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Korean J Parasitol > Volume 34(1):1996 > Article

Original Article
Korean J Parasitol. 1996 Mar;34(1):79-85. English.
Published online Mar 20, 1996.  http://dx.doi.org/10.3347/kjp.1996.34.1.79
Copyright © 1996 by The Korean Society for Parasitology
Biochemical and molecular characterization of a strain KA/S2 of Acanthamoeba castellanii isolated from Korean soil
Dong-Il Chung,*1Hyun-Hee Kong,1Hak-Sun Yu,1Yu-Mi Oh,1Sung-Tae Yee,2 and Young-Jin Lim2
1Department of Parasitology, Kyungpook National University School of Medicine, Taegu 700-422, Korea.
2Department of Parasitology, college of Medicine, Dong-A University, Pusan 602-103, Korea.
Received October 20, 1995; Accepted January 03, 1996.


A strain, KA/S2, isolated from Korean soil and morphologically assigned to Acanthamoeba castellanii, was characterized by isoenzyme analysis, and total proteins profile, and mitochondrial (Mt) DNA restriction fragment length polymorphism (RFLP), and compared with four reference strains assigned to the species (the authenitic Castellani, Neff, Ma, and Chang strains). It was found that four isoenzyme, total proteins, and Mt DNA RFLP patterns by eight restriction endonucleases of the strain KA/S2 were identical with those of the Neff strain, isolated from soil of California, USA. The Chang strain was unique in its morphology and total protein patterns. Interstrain polymorphisms of isoenzyme profiles and Mt DNA RFLP patterns were observed among the Castellani, Neff, Ma, and Chang strains. Mt DNA RFLP was confirmed to be highly appropriate for the strain characterization and identification of Acanthamoeba spp.


Fig. 1
Photomicrographs of the cysts of Acanthamoeba castellanii. A. KA/S2 strain; B. Castellani strain; C. Neff strain; D. Ma strain; E. Chang strain. Bars indicate 10 µm.

Fig. 2
Zymograms for isoenzymes of KA/S2 and reference strains of A. Castellanii separated by polyacrylamide gel isoelectric focusing in pH gradient 3~10. A. Acid phosphatase; B. Leucine amino peptidase; C. Glucose-6-phosphate dehydrogenase; D. Glucose phosphate Isomerase. Numbers above lanes refer to the strains listed in Table 2.

Fig. 3
Total proteins of Acanthamoeba separated by polyacrylamide gel isoelectric focusing in pH gradient 3-7. Numbers above lanes refer to the strains listed in Table 2.

Fig. 4
Agarose gel electrophoretic fingerprints of mitochondrial DNA from KA/S2 and reference strains of A. Castellanii. A.EcoR I digests; B.Hpa I digests; C.Bgl II digests; D.Sca I digests; E.Cla I digests; F.Xba I digests; G.Sst I digests; H.Sal I digests. Numbers above lanes refer to the strains listed in Table 2. Size marker is Hind III digested λ phage DNA (M).


Table 1
Morphology of Acanthamoeba castellanii KA/S2 and the reference strains

Table 2
Development conditions of enzymes tested (final concentration/100ml)

1. Bogler SA, Zarley CD, Burianek LL, Fuerst PA, Byers TJ. Interstrain mitochondrial DNA polymorphism detected in Acanthamoeba by restriction endonuclease analysis. Mol Biochem Parasitol 1983;8(2):145–163.
2. Burger G, Plante I, Lonergan KM, Gray MW. The mitochondrial DNA of the amoeboid protozoon, Acanthamoeba castellanii: complete sequence, gene content and genome organization. J Mol Biol 1995;245(5):522–537.
3. Byers TJ, et al. J Protozool 1983;30:198–203.
4. Byers TJ, et al. J Protozool 1990;37(4):17s–25s.
6. Douglas M. J Trop Med London 1930;33:258–259.
7. Gautom RK, Lory S, Seyedirashti S, Bergeron DL, Fritsche TR. Mitochondrial DNA fingerprinting of Acanthamoeba spp. isolated from clinical and environmental sources. J Clin Microbiol 1994;32(4):1070–1073.
8. Gunderson JH, Sogin ML. Length variation in eukaryotic rRNAs: small subunit rRNAs from the protists Acanthamoeba castellanii and Euglena gracilis. Gene 1986;44(1):63–70.
9. Ferris SD, Wilson AC, Brown WM. Evolutionary tree for apes and humans based on cleavage maps of mitochondrial DNA. Proc Natl Acad Sci U S A 1981;78(4):2432–2436.
10. Kilvington S, Beeching JR, White DG. Differentiation of Acanthamoeba strains from infected corneas and the environment by using restriction endonuclease digestion of whole-cell DNA. J Clin Microbiol 1991;29(2):310–314.
11. Kong HH, Park JH, Chung DI. Interstrain polymorphisms of isoenzyme profiles and mitochondrial DNA fingerprints among seven strains assigned to Acanthamoeba polyphaga. Korean J Parasitol 1995;33(4):331–340.
12. Ma P, Willaert E, Juechter KB, Stevens AR. A case of keratitis due to Acanthamoeba in New York, New York, and features of 10 cases. J Infect Dis 1981;143(5):662–667.
13. McLaughlin GL, Brandt FH, Visvesvara GS. Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae. J Clin Microbiol 1988;26(9):1655–1658.
14. MacKay RM, Doolittle WF. Nucleotide sequences of Acanthamoeba castellanii 5S and 5.8S ribosomal ribonucleic acids: phylogenetic and comparative structural analyses. Nucleic Acids Res 1981;9(14):3321–3334.
15. Neff RJ. J Protozool 1957;4:176–182.
16. Page FC. Re-definition of the genus Acanthamoeba with descriptions of three species. J Protozool 1967;14(4):709–724.
18. Vodkin MH, Howe DK, Visvesvara GS, McLaughlin GL. Identification of Acanthamoeba at the generic and specific levels using the polymerase chain reaction. J Protozool 1992;39(3):378–385.
19. Yagita K. Jpn J Parasitol 1993;42:468–478.
20. Yagita K, Endo T. Restriction enzyme analysis of mitochondrial DNA of Acanthamoeba strains in Japan. J Protozool 1990;37(6):570–575.
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