Skip to main content

Identification of Legionella in the Environment

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 954))

Abstract

Legionella is ubiquitous in freshwater systems worldwide and can also be found in soil. Legionellosis may be caused by inhalation of aerosolized water or soil particles containing Legionella. Isolation of Legionella from the environment is an essential step in outbreak investigation and may also be performed within the context of a hazard analysis and control risk management plan. Culture remains the gold standard for detection of Legionella in environmental samples. Specific properties of environmental sites that could be a source of Legionella contamination, collection of samples from such sites, and procedures for culture of these samples for Legionella are described in this chapter.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Fields BS, Benson RF, Besser RE (2002) Legionella and Legionnaires’ disease: 25 years of investigation. Clin Microbiol Rev 15:506–526

    Article  PubMed  Google Scholar 

  2. Anonymous (2000) Minimizing the risk of legionellosis associated with building water systems. American Society of Heating, Refrigerating, and Air-conditioning Engineers, Inc., Atlanta

    Google Scholar 

  3. Casati S, Gioria-Martinoni A, Gaia V (2009) Commercial potting soils as an alternative infection source of Legionella pneumophila and other Legionella species in Switzerland. Clin Microbiol Infect 15:571–575

    Article  PubMed  CAS  Google Scholar 

  4. den Boer JW, Yzerman EP, Jansen R, Bruin JP, Verhoef LP, Neve G, van der Zwaluw K (2007) Legionnaires’ disease and gardening. Clin Microbiol Infect 13:88–91

    Article  Google Scholar 

  5. Hughes MS, Steele TW (1994) Occurrence and distribution of Legionella species in composted plant materials. Appl Environ Microbiol 60:2003–2005

    PubMed  CAS  Google Scholar 

  6. Koide M, Saito A, Okazaki M, Umeda B, Benson RF (1999) Isolation of Legionella longbeachae serogroup 1 from potting soils in Japan. Clin Infect Dis 29:943–944

    Article  PubMed  CAS  Google Scholar 

  7. Pravinkumar SJ, Edwards G, Lindsay D, Redmond S, Stirling J, House R, Kerr J, Anderson E, Breen D, Blatchford O, McDonald E, Brown A (2010) A cluster of Legionnaires’ disease caused by Legionella longbeachae linked to potting compost in Scotland, 2008–2009. Euro Surveill 15:19496

    PubMed  CAS  Google Scholar 

  8. Travis TC, Brown EW, Peruski LF, Siludjai D, Jorakate P, Salika P, Yang G, Kozak NA, Kodani M, Warner AK, Lucas CE, Thurman KA, Winchell JM, Thamthitiwat S, and Fields BS (2012) Survey of Legionella species found in Thai soil, International journal of microbiology 2012, 218791.

    Google Scholar 

  9. Velonakis EN, Kiousi IM, Koutis C, Papadogiannakis E, Babatsikou F, Vatopoulos A (2010) First isolation of Legionella species, including L. pneumophila serogroup 1, in Greek potting soils: possible importance for public health. Clin Microbiol Infect 16:763–766

    Article  PubMed  CAS  Google Scholar 

  10. Atlas RM (1999) Legionella: from environmental habitats to disease pathology, detection and control. Environ Microbiol 1:283–293

    Article  PubMed  CAS  Google Scholar 

  11. Bugler TJL, Fields BS, Miller RD (2010) Cooling towers, drift, and legionellosis. CTI J 31:30–47

    Google Scholar 

  12. Alary M, Joly JR (1992) Comparison of culture methods and an immunofluorescence assay for the detection of Legionella pneumophila in domestic hot water devices. Curr Microbiol 25:19–23

    Article  PubMed  CAS  Google Scholar 

  13. Flournoy DJ, Belobraydic KA, Silberg SL, Lawrence CH, Guthrie PJ (1988) False positive Legionella pneumophila direct immunofluorescent monoclonal antibody test caused by bacillus cereus spores. Diagn Microbiol Infect Dis 9:123–125

    Article  PubMed  CAS  Google Scholar 

  14. Tronel H, Hartemann P (2009) Overview of diagnostic and detection methods for legionellosis and Legionella spp. Lett Appl Microbiol 48:653–656

    PubMed  CAS  Google Scholar 

  15. Anonymous (2005) Procedures for the recovery of Legionella from the environment, (Fields, B. S., Ed.), US Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, Atlanta, GA

    Google Scholar 

  16. Barbaree JM, Gorman GW, Martin WT, Fields BS, Morrill WE (1987) Protocol for sampling environmental sites for legionellae. Appl Environ Microbiol 53:1454–1458

    PubMed  CAS  Google Scholar 

  17. Miller RP (1979) Cooling towers and evaporative condensers. Ann Intern Med 90:667–670

    PubMed  CAS  Google Scholar 

  18. Anonymous (2009) Cooling Tower Fundamentals, (Hensley, J. C., Ed.) 2nd ed., SPX Cooling Technologies, Inc., Overland Park, KS.

    Google Scholar 

  19. O’Loughlin RE, Kightlinger L, Werpy MC, Brown E, Stevens V, Hepper C, Keane T, Benson RF, Fields BS, Moore MR (2007) Restaurant outbreak of Legionnaires’ disease associated with a decorative fountain: an environmental and case-control study. BMC Infect Dis 7:93

    Article  PubMed  Google Scholar 

  20. Hlady WG, Mullen RC, Mintz CS, Shelton BG, Hopkins RS, Daikos GL (1993) Outbreak of Legionnaire’s disease linked to a decorative fountain by molecular epidemiology. Am J Epidemiol 138:555–562

    PubMed  CAS  Google Scholar 

  21. Woo AH, Goetz A, Yu VL (1992) Transmission of Legionella by respiratory equipment and aerosol generating devices. Chest 102:1586–1590

    Article  PubMed  CAS  Google Scholar 

  22. Mastro TD, Fields BS, Breiman RF, Campbell J, Plikaytis BD, Spika JS (1991) Nosocomial legionnaires’ disease and use of medication nebulizers. J Infect Dis 163:667–671

    Article  PubMed  CAS  Google Scholar 

  23. Macfarlane JT (1983) Legionnaires’ disease: update. Br Med J (Clin Res Ed) 287:443–444

    Article  CAS  Google Scholar 

  24. Stout JE, Yu VL, Muraca P (1985) Isolation of Legionella pneumophila from the cold water of hospital ice machines: implications for origin and transmission of the organism. Infect Control 6:141–146

    PubMed  CAS  Google Scholar 

  25. Mahoney FJ, Hoge CW, Farley TA, Barbaree JM, Breiman RF, Benson RF, McFarland LM (1992) Communitywide outbreak of Legionnaires’ disease associated with a grocery store mist machine. J Infect Dis 165:736–739

    Article  PubMed  CAS  Google Scholar 

  26. Wallensten A, Oliver I, Ricketts K, Kafatos G, Stuart JM, Joseph C (2010) Windscreen wiper fluid without added screen wash in motor vehicles: a newly identified risk factor for Legionnaires’ disease. Eur J Epidemiol 25:661–665

    Article  PubMed  Google Scholar 

  27. Boulanger CA, Edelstein PH (1995) Precision and accuracy of recovery of Legionella pneumophila from seeded tap water by filtration and centrifugation. Appl Environ Microbiol 61:1805–1809

    PubMed  CAS  Google Scholar 

  28. Ta AC, Stout JE, Yu VL, Wagener MM (1995) Comparison of culture methods for monitoring Legionella species in hospital potable water systems and recommendations for standardization of such methods. J Clin Microbiol 33:2118–2123

    PubMed  CAS  Google Scholar 

  29. Bopp CA, Sumner JW, Morris GK, Wells JG (1981) Isolation of Legionella spp. from environmental water samples by low-pH treatment and use of a selective medium. J Clin Microbiol 13:714–719

    PubMed  CAS  Google Scholar 

  30. Amemura-Maekawa J, Hayakawa Y, Sugie H, Moribayashi A, Kura F, Chang B, Wada A, Watanabe H (2004) Legioliulin, a new isocoumarin compound responsible for blue-white autofluorescence in Legionella (Fluoribacter) dumoffii under long-wavelength UV light. Biochem Biophys Res Commun 323:954–959

    Article  PubMed  CAS  Google Scholar 

  31. Sanden GN, Morrill WE, Fields BS, Breiman RF, Barbaree JM (1992) Incubation of water samples containing amoebae improves detection of legionellae by the culture method. Appl Environ Microbiol 58:2001–2004

    PubMed  CAS  Google Scholar 

  32. Wiedenmann A, Langhammer W, Botzenhart K (2001) A case report of false negative Legionella test results in a chlorinated public hot water distribution system due to the lack of sodium thiosulfate in sampling bottles. Int J Hyg Environ Health 204:245–249

    Article  PubMed  CAS  Google Scholar 

  33. Feeley JC, Gibson RJ, Gorman GW, Langford NC, Rasheed JK, Mackel DC, Baine WB (1979) Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila. J Clin Microbiol 10:437–441

    PubMed  CAS  Google Scholar 

  34. Edelstein PH, Finegold SM (1979) Use of a semiselective medium to culture Legionella pneumophila from contaminated lung specimens. J Clin Microbiol 10:141–143

    PubMed  CAS  Google Scholar 

  35. Wadowsky RM, Yee RB (1981) Glycine-containing selective medium for isolation of Legionellaceae from environmental specimens. Appl Environ Microbiol 42:768–772

    PubMed  CAS  Google Scholar 

  36. Morrill WE, Barbaree JM, Fields BS, Sanden GN, Martin WT (1990) Increased recovery of Legionella micdadei and Legionella bozemanii on buffered charcoal yeast extract agar supplemented with albumin. J Clin Microbiol 28:616–618

    PubMed  CAS  Google Scholar 

  37. Orrison LH, Cherry WB, Tyndall RL, Fliermans CB, Gough SB, Lambert MA, McDougal LK, Bibb WF, Brenner DJ (1983) Legionella oakridgensis: unusual new species isolated from cooling tower water. Appl Environ Microbiol 45:536–545

    PubMed  CAS  Google Scholar 

  38. Lucas CE, Taylor TH Jr, Fields BS (2011) Accuracy and precision of Legionella isolation by US laboratories in the ELITE program pilot study. Water Res 45:4428–4436

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

We thank Ellen W. Brown and Joseph C. Carpenter for their help in preparation of the chapter.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Natalia A. Kozak .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Kozak, N.A., Lucas, C.E., Winchell, J.M. (2013). Identification of Legionella in the Environment. In: Buchrieser, C., Hilbi, H. (eds) Legionella. Methods in Molecular Biology, vol 954. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-161-5_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-161-5_1

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-160-8

  • Online ISBN: 978-1-62703-161-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics