Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case

  • Yu, Hak-Sun (Department of Parasitology, Pusan National University School of Medicine) ;
  • Jeong, Hae-Jin (Department of Parasitology, Pusan National University School of Medicine) ;
  • Hong, Yeon-Chul (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Seol, Seong-Yong (Department of Microbiology, Kyungpook National University School of Medicine) ;
  • Chung, Dong-Il (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Kong, Hyun-Hee (Department of Parasitology, Kyungpook National University School of Medicine)
  • Published : 2007.03.31

Abstract

Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multi-plied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

Keywords

References

  1. Ahn GS, Choi EY, Jeon KW (1990) A Symbiosome-membrane specific protein in symbiont-bearing Amoeba proteus as studied with monoclonal antibody. Endocytobios. Cell Res 7: 45-50
  2. Amer AO, Swanson MS (2002) A phagosome of one's own: a microbial guide to life in the macrophage. Curr Opin Microbiol 5: 56-6I https://doi.org/10.1016/S1369-5274(02)00286-2
  3. Amano A, Nakagawa I, Yoshimori T (2006) Autophagy in innate immunity against intracellular bacteria. J Biochem (Tokyo) 140: 161-166 https://doi.org/10.1093/jb/mvj162
  4. Burger G, Plante I, Lonergan KM, Gray MW (1995) The mitochondrial DNA of the amoeboid protozoon, Acanthamoeba castellanii: complete sequence, gene content and genome organization. J Mol BioI 245: 522-537 https://doi.org/10.1006/jmbi.1994.0043
  5. Choi JY, Lee TW, Jeon KW, Ahn TI (1997) Evidence for symbiont- induced alteration of a host's expression: Irreversible loss of SAM synthetase from Ameba proteus. J Eukaryot Microbiol 44: 412-419 https://doi.org/10.1111/j.1550-7408.1997.tb05717.x
  6. Chung DI, Kong HH, Kim TH, Hwang MY, Yu HS, Yun HC, Seol SY (1997) Bacterial endosymbiosis within the cytoplasm of Acanthamoeba lugdunensis isolated from a contact lens storage case. Korean J Parasitol 35: 127-133 https://doi.org/10.3347/kjp.1997.35.2.127
  7. Cirillo JD, Falkow S, Tompkins LS (1994) Growth of Leigionella pneumophila in Acanthamoeba castellanii enhances invasion. Infect Immun 62: 3254-326I
  8. Cirillo JD, Falkow S, Tompkins LS, Bermudez LE (1997) Interaction of Mycobacterium avium with environmental amoebae enhances virulence. Infect Immun 65: 3759- 3767
  9. Felsenstein J (1993) PHYLIP manual, version 3.5. Department of Genetics, University of Washington
  10. Fields BS (1996) The molecular ecology of Legionellae. Trends Microbiol 4: 286-290 https://doi.org/10.1016/0966-842X(96)10041-X
  11. Holden EP, Winkler HH, Wood DO, Leinbach ED (1984) Intracellular growth of Legionella pneumophila within Acanthamoeba castellanii Neff. Infect Immun 45: 18-24
  12. Jadin JB (1973) Hypotheses on the adaption of amoebas of the limax group to man and animals. Ann Parasitol Hum Comp 48: 199-204 https://doi.org/10.1051/parasite/1973482199
  13. Khan NA (2006) Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev 30: 564-595 https://doi.org/10.1111/j.1574-6976.2006.00023.x
  14. Kilvington 5, Price J (1990) Survival of Legionella pneumophila within cysts of Acanthamoeba polyphaga following chlorine exposure. J Appl Bacteriol 68: 519-525 https://doi.org/10.1111/j.1365-2672.1990.tb02904.x
  15. Kong HH, Chung OI (1996) Bacterial endosymbionts of Acanthamoeba sp. isolated from cooling tower water. Jpn J Parasitol 45: 505-51I
  16. Ly TM, Muller HE (1990) Ingested Listeria monocytogenes survive and multiply in protozoa. J Med Microbiol 33: 51- 54 https://doi.org/10.1099/00222615-33-1-51
  17. Miltner EC, Bermudez LE (2000) Mycobacterium avium growth in Acanthamoeba castellanii is protected from the effects of antimicrobials. Antimicrob Agents Chemother 44: 1990-1994 https://doi.org/10.1128/AAC.44.7.1990-1994.2000
  18. Park MS (1990) Nucleotide sequence of a symbiotic bacterial gene coding for a protein used by the host Amoeba proteius. Endocytobios. Cell Res 7: 37-44
  19. Reed C, von Reyn CF, Chamblee 5, Ellerbrock TV, Johnson JW, Marsh BJ,Johnson LS, Trenschel RJ, Horsburgh CR Jr (2006) Environmental risk factors for infection with Mycobacterium avium complex. Am J Epidemiol 164: 32-40 https://doi.org/10.1093/aje/kwj159
  20. Rosenberger CM, Finaly BB (2003) Phagocyte sabotage: disruption of macrophage signaling by bacterial pathogens. Nat Rev Mol Cell BioI 4: 385-396 https://doi.org/10.1038/nrm1104
  21. Shin HI, Cho MS, Kim HI, Im KI (1999) Isoenzyme patterns and phylogenetic relationships in Acanthamoeba spp. isolated from contact lens containers in Korea. Korean J Parasitol 37: 229-236 https://doi.org/10.3347/kjp.1999.37.4.229
  22. Smith I (2003) Mycobacterium tuberculosis pathogenesis and molecular determinants of virulence. Clin Microbiol Rev 16: 463-496 https://doi.org/10.1128/CMR.16.3.463-496.2003
  23. Steinert M, Birkness K, White E, Fields B, Quinn F (1998) Mycobacterium avium bacilli grow saprozoically in coculture with Acanthamoeba polyphaga and survive within cyst walls. Appl Environ Microbiol 64: 2256-2261
  24. Taylor GM, Stewart GR, Cooke M, Chaplin 5, Ladva 5, Kirkup I, Palmer 5, Young DB (2003) Koch's bacillus- a look at the first isolate of Mycobacterium tuberculosis from a modern perspective. Micorbiology 149: 3213-3220
  25. Thorn 5, Warhurst D, Drasar BS (1992) Association of Vibrio cholerae with fresh water amoebae. J Med Microbiol 36: 303-306 https://doi.org/10.1099/00222615-36-5-303
  26. Winiecka-Krusnell J, Linder E (2001) Bacterial infections of free-living amoebae. Res Microbiol 152: 613-619 https://doi.org/10.1016/S0923-2508(01)01240-2
  27. Yagita K, Endo T (1990) Restriction enzyme analysis of mito- chondrial DNA of Acanthamoeba strains in Japan. J Protozool 37: 570-575 https://doi.org/10.1111/j.1550-7408.1990.tb01267.x
  28. Yu HS, Kong HH, Kim SY, Hahn YH, Hahn TW, Chung DI (2004) Laboratory investigation of Acanthamoeba lugdunensis from patients with keratitis. Invest Ophthalmol Vis Sci 45: 1418-1426 https://doi.org/10.1167/iovs.03-0433