Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Molecular characterization of bacterial endosymbionts of Acanthamoeba isolates from infected corneas of Korean patients

  • Xuan, Ying-Hua (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Yu, Hak-Sun (Department of Parasitology, Pusan National University School of Medicine) ;
  • Jeong, Hae-Jin (Department of Parasitology, Pusan National University School of Medicine) ;
  • Seol, Sung-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
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Abstract

The endosymbionts of 4 strains of Acanthamoeba(KA/E9, KA/E21, KA/E22, and KA/E23) isolated from the infected corneas of Korean patients were characterized via orcein stain, transmission electron microscopic examination, and 16S rDNA sequence analysis. Double membrane-bound, rod-shaped endosymbionts were distributed randomly throughout both the trophozoites and cysts of each of Acanthamoeba isolates. The endosymbionts of KA/E9, KA/E22, and KA/E23 were surrounded by electron-translucent areas. No lacunae-like structures were observed in the endosymbionts of KA/E21, the bacterial cell walls of which were studded with host ribosomes. Comparative analyses of the 16S rDNA sequences showed that the endosymbionts of KA/E9, KA/E22 and KA/E23 were closely related to Caedibacter caryophilus, whereas the KA/E21 endosymbiont was assigned to the Cytophaga-Flavobacterium-Bacteroides(CFB) phylum. In the 4 strains of Acanthamoeba, the hosts of the endosymbionts were identified as belonging to the Acanthamoeba castellanii complex, which corresponds to the T4 genotype. Acanthamoeba KA/E21 evidenced characteristics almost identical to those of KA/E6, with the exception of the existence of endosymbionts. The discovery of these endosymbionts from Acanthamoeba may prove essential to future studies focusing on interactions between the endosymbionts and the amoebic hosts.

Keywords

References

  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol BioI 215: 403-410 https://doi.org/10.1016/S0022-2836(05)80360-2
  2. Barker J, Humphrey TJ, Brown MW (1999) Survival of Escherichia coli 0157 in a soil protozoan: implications for disease. FEMS Microbial Lett 173: 291-295 https://doi.org/10.1111/j.1574-6968.1999.tb13516.x
  3. Beier CL, Horn M, Michel R, Schweikert M, Gortz HD, Wagner M (2002) The genus Caedibacter comprises endosymbionts of Paramecium spp. related to the Rickettsiales (Alphaproteobacteria) and to Francisella tularensis (Gammaproteobacteria). Appl Environ Microbial 68: 6043-6050 https://doi.org/10.1128/AEM.68.12.6043-6050.2002
  4. Burger G, Plante I, Lonergan KM, Gray MW (1995) The mitochondrial DNA of 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. Cengiz AM, Harmis N, Stapleton F (2000) Co-incubation of Acanthamoeba castellanii with strains of Pseudomonas aeruginosa alters the survival of amoeba. Clin Experiment Ophthalmol 28: 191-193 https://doi.org/10.1046/j.1442-9071.2000.00291.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, Bermudez LE (1997) Interaction of Mycobacterium avium with environmental amoebae enhances virulence. Infect Immun 65: 3759- 3767
  8. Essig A, Heinemann M, Simnacher V, Marre R (1997) Infection of Acanthamoeba castellanii by Chlamydia pneumoniae. Appl Environ Microbiol 63: 1396-1399
  9. Fritsche TR, Gautom RK, Seyedirashti S, Bergeron DL, Lindquist TD (1993) Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J Clin Microbiol 31: 1122-1126
  10. Gunderson JH, Sogin ML (1986) Length variation in eukaryotic rRNAs: small subunit rRNAs from the protists Acanthamoeba castellanii and Euglena gracilis. Gene 44: 63- 70 https://doi.org/10.1016/0378-1119(86)90043-0
  11. Hall J, Voelz H (1985) Bacterial endosymbionts of Acanthamoeba sp. J Parasitol 71: 89-95 https://doi.org/10.2307/3281982
  12. Horn M, Fritsche TR, Gautom RK, Schleifer KH, Wagner M (1999) Novel bacterial endosymbionts of Acanthamoeba spp. related to the Paramecium caudatum symbiont Caedibacter caryophilus. Environ Microbiol 1: 357-367 https://doi.org/10.1046/j.1462-2920.1999.00045.x
  13. Horn M, Harzenetter MD, Linner T, Schmid EN, Muller KD, Michel R, Wagner M (2001) Members of the Cytophaga- Flavobacterium-Bacteroides phylum as intracellular bacteria of Acanthamoebae: proposal of 'Candidatus Amoebophilus asiaticus'. Environ Microbiol 3: 440-449 https://doi.org/10.1046/j.1462-2920.2001.00210.x
  14. Jones BR, McGill JI, Steele AD (1975) Recurrent suppurative kerato-uveitis with loss of eye due to infection by Acanthamoeba castellani. Trans Ophthalmol Sac U K 95: 210-213
  15. Kong HH, Chung DI (1996) PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga. Korean J Parasitol34: 127-134 https://doi.org/10.3347/kjp.1996.34.2.127
  16. Kong HH, Shin JY, Yu HS, Kim J, Hahn TW, Hahn YH, Chung DI (2002) Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J Clin Microbiol 40: 1199-1206 https://doi.org/10.1128/JCM.40.4.1199-1206.2002
  17. Lee SM, Choi YJ, Ryu HW, Kong HH, Chung DI (1997) Species identification and molecular characterization of Acanthamoeba isolated from contact lens paraphernalia. Korean J Ophthalmol 11: 39-50 https://doi.org/10.3341/kjo.1997.11.1.39
  18. Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16: 273-307 https://doi.org/10.1128/CMR.16.2.273-307.2003
  19. Rowbotham TJ (1986) Current views on the relationship between amoebae, legionellae and man. Isr J Med Sci 22: 678-689
  20. Rodriguez-Zaragoza S (1994) Ecology of free living amoebae. Crit Rev Microbiol 20: 225-241 https://doi.org/10.3109/10408419409114556
  21. Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol44: 846-849 https://doi.org/10.1099/00207713-44-4-846
  22. 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
  23. Stothard DR, Schroeder-Diedrich JM, Awwad MH, Gast RJ, Ledee DR, Rodriguez-Zaragoza S, Dean CL, Fuerst PA, Byers TJ (1998) The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45: 45- 54 https://doi.org/10.1111/j.1550-7408.1998.tb05068.x
  24. Thomspon JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL-X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 4876-4882 https://doi.org/10.1093/nar/25.24.4876
  25. Winiecka-Krusnell J, Linder E (2001) Bacterial infections of free-living amoebae. Res Microbiol152: 613-619 https://doi.org/10.1016/S0923-2508(01)01240-2
  26. Yagita K, Endo T (1990) Restriction enzyme analysis of mitochondrial DNA of Acanthamoeba strains in Japan. J Protozool 37: 570-575 https://doi.org/10.1111/j.1550-7408.1990.tb01267.x
  27. Yu HS, Choi KH, Kim HK, Kong HH, Chung DI (2001) Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Korean J Parasitol39: 161-170 https://doi.org/10.3347/kjp.2001.39.2.161