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. 1992 Jun;58(6):2001–2004. doi: 10.1128/aem.58.6.2001-2004.1992

Incubation of water samples containing amoebae improves detection of legionellae by the culture method.

G N Sanden 1, W E Morrill 1, B S Fields 1, R F Breiman 1, J M Barbaree 1
PMCID: PMC195717  PMID: 1622278

Abstract

Some protozoans isolated from aquatic habitats, including domestic water supplies, can support the intracellular replication of autochthonous legionellae in vitro. We studied the effect of incubating water samples containing amoebae on the sensitivity of culture for legionellae. Samples collected during investigations of legionellosis epidemics and shown by conventional culture procedures to contain amoebae, but not legionellae, were incubated at 35 degrees C and replated. Legionellae were recovered from 59 of 144 such samples. Species isolated included L. pneumophila, L. anisa, L. bozemanii, L. gormanii, L. micdadei, L. rubrilucens, L. sainthelensi, L. steigerwaltii, and an unnamed species. Acanthamoeba polyphaga, Acanthamoeba hatchetti, a Rosculus sp., Hartmannella vermiformis, and Vahlkampfia spp. were among the autochthonous amoebae identified. Legionellae were recovered by this procedure from only 3 of 63 samples that were negative for amoebae by conventional culture procedures. These results show that water samples negative for legionellae, but positive for amoebae, by standard culture techniques should be incubated and replated to maximize the sensitivity of culture for legionellae.

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Selected References

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  1. Anand C. M., Skinner A. R., Malic A., Kurtz J. B. Interaction of L. pneumophilia and a free living amoeba (Acanthamoeba palestinensis). J Hyg (Lond) 1983 Oct;91(2):167–178. doi: 10.1017/s0022172400060174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbaree J. M., Fields B. S., Feeley J. C., Gorman G. W., Martin W. T. Isolation of protozoa from water associated with a legionellosis outbreak and demonstration of intracellular multiplication of Legionella pneumophila. Appl Environ Microbiol. 1986 Feb;51(2):422–424. doi: 10.1128/aem.51.2.422-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breiman R. F., Fields B. S., Sanden G. N., Volmer L., Meier A., Spika J. S. Association of shower use with Legionnaires' disease. Possible role of amoebae. JAMA. 1990 Jun 6;263(21):2924–2926. [PubMed] [Google Scholar]
  4. Brown A., Yu V. L., Magnussen M. H., Vickers R. M., Garrity G. M., Elder E. M. Isolation of Pittsburgh pneumonia agent from a hospital shower. Appl Environ Microbiol. 1982 Mar;43(3):725–726. doi: 10.1128/aem.43.3.725-726.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dowling J. N., Pasculle A. W., Frola F. N., Zaphyr M. K., Yee R. B. Infections caused by Legionella micdadei and Legionella pneumophila among renal transplant recipients. J Infect Dis. 1984 May;149(5):703–713. doi: 10.1093/infdis/149.5.703. [DOI] [PubMed] [Google Scholar]
  6. Feeley J. C., Gorman G. W., Weaver R. E., Mackel D. C., Smith H. W. Primary isolation media for Legionnaires disease bacterium. J Clin Microbiol. 1978 Sep;8(3):320–325. doi: 10.1128/jcm.8.3.320-325.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fields B. S., Barbaree J. M., Sanden G. N., Morrill W. E. Virulence of a Legionella anisa strain associated with Pontiac fever: an evaluation using protozoan, cell culture, and guinea pig models. Infect Immun. 1990 Sep;58(9):3139–3142. doi: 10.1128/iai.58.9.3139-3142.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fields B. S., Nerad T. A., Sawyer T. K., King C. H., Barbaree J. M., Martin W. T., Morrill W. E., Sanden G. N. Characterization of an axenic strain of Hartmannella vermiformis obtained from an investigation of nosocomial legionellosis. J Protozool. 1990 Nov-Dec;37(6):581–583. doi: 10.1111/j.1550-7408.1990.tb01269.x. [DOI] [PubMed] [Google Scholar]
  9. Fields B. S., Shotts E. B., Jr, Feeley J. C., Gorman G. W., Martin W. T. Proliferation of Legionella pneumophila as an intracellular parasite of the ciliated protozoan Tetrahymena pyriformis. Appl Environ Microbiol. 1984 Mar;47(3):467–471. doi: 10.1128/aem.47.3.467-471.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fisher-Hoch S. P., Bartlett C. L., Tobin J. O., Gillett M. B., Nelson A. M., Pritchard J. E., Smith M. G., Swann R. A., Talbot J. M., Thomas J. A. Investigation and control of an outbreaks of legionnaires' disease in a district general hospital. Lancet. 1981 Apr 25;1(8226):932–936. doi: 10.1016/s0140-6736(81)91626-3. [DOI] [PubMed] [Google Scholar]
  11. Fitzgeorge R. B., Dennis P. J. Isolation of Legionella pneumophila from water supplies: comparison of methods based on the guinea-pig and culture media. J Hyg (Lond) 1983 Oct;91(2):179–187. doi: 10.1017/s0022172400060186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fliermans C. B., Cherry W. B., Orrison L. H., Smith S. J., Tison D. L., Pope D. H. Ecological distribution of Legionella pneumophila. Appl Environ Microbiol. 1981 Jan;41(1):9–16. doi: 10.1128/aem.41.1.9-16.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Henke M., Seidel K. M. Association between Legionella pneumophila and amoebae in water. Isr J Med Sci. 1986 Sep;22(9):690–695. [PubMed] [Google Scholar]
  14. Morrill W. E., Barbaree J. M., Fields B. S., Sanden G. N., Martin W. T. Increased recovery of Legionella micdadei and Legionella bozemanii on buffered charcoal yeast extract agar supplemented with albumin. J Clin Microbiol. 1990 Mar;28(3):616–618. doi: 10.1128/jcm.28.3.616-618.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Neill M. A., Gorman G. W., Gibert C., Roussel A., Hightower A. W., McKinney R. M., Broome C. V. Nosocomial legionellosis, Paris, France. Evidence for transmission by potable water. Am J Med. 1985 Apr;78(4):581–588. doi: 10.1016/0002-9343(85)90399-7. [DOI] [PubMed] [Google Scholar]
  16. Newsome A. L., Baker R. L., Miller R. D., Arnold R. R. Interactions between Naegleria fowleri and Legionella pneumophila. Infect Immun. 1985 Nov;50(2):449–452. doi: 10.1128/iai.50.2.449-452.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Roszak D. B., Colwell R. R. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. doi: 10.1128/mr.51.3.365-379.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rowbotham T. J. Current views on the relationships between amoebae, legionellae and man. Isr J Med Sci. 1986 Sep;22(9):678–689. [PubMed] [Google Scholar]
  19. Rowbotham T. J. Isolation of Legionella pneumophila from clinical specimens via amoebae, and the interaction of those and other isolates with amoebae. J Clin Pathol. 1983 Sep;36(9):978–986. doi: 10.1136/jcp.36.9.978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rowbotham T. J. Preliminary report on the pathogenicity of Legionella pneumophila for freshwater and soil amoebae. J Clin Pathol. 1980 Dec;33(12):1179–1183. doi: 10.1136/jcp.33.12.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Spriggs D. R. Legionella, microbial ecology, and inconspicuous consumption. J Infect Dis. 1987 May;155(5):1086–1087. doi: 10.1093/infdis/155.5.1086. [DOI] [PubMed] [Google Scholar]
  22. Tison D. L., Seidler R. J. Legionella incidence and density in potable drinking water supplies. Appl Environ Microbiol. 1983 Jan;45(1):337–339. doi: 10.1128/aem.45.1.337-339.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wadowsky R. M., Butler L. J., Cook M. K., Verma S. M., Paul M. A., Fields B. S., Keleti G., Sykora J. L., Yee R. B. Growth-supporting activity for Legionella pneumophila in tap water cultures and implication of hartmannellid amoebae as growth factors. Appl Environ Microbiol. 1988 Nov;54(11):2677–2682. doi: 10.1128/aem.54.11.2677-2682.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wadowsky R. M., Yee R. B., Mezmar L., Wing E. J., Dowling J. N. Hot water systems as sources of Legionella pneumophila in hospital and nonhospital plumbing fixtures. Appl Environ Microbiol. 1982 May;43(5):1104–1110. doi: 10.1128/aem.43.5.1104-1110.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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