Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81
Warning: fopen(upload/ip_log/ip_log_2024-11.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83
Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84 Close relatedness of Acanthamoeba pustulosa with Acanthamoeba palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns
Close relatedness of Acanthamoeba pustulosa with Acanthamoeba palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns
Young Ho Kim,1Mee Sun Ock,2Ho Cheol Yun,1Mee Yul Hwang,1Hak Sun Yu,1Hyun Hee Kong,1 and Dong Il Chung*1
1Department of Parasitology, Kyungpook National University School of Medicine, Taegu 700-422, Korea.
2Department of Parasitology, College of Medicine, Kosin University, Pusan 602-030, Korea.
Received September 06, 1996; Accepted November 29, 1996.
Abstract
The taxonomic validity of morphological group III Acanthamoeba spp. is uncertain. In the present study, six type strains of group III Acanthamoeba spp., Acanthamoeba culbertsoni, A. healyi, A. pustulosa, A. palestinensis, A. royreba and A. lenticulata were subjected for the evaluation of their taxonomic validity by comparison of the isoenzyme patterns by isoelectic focusing on polyacrylamide gels, mitochondrial DNA (Mt DNA) restriction fragment length polymorphism (RFLP), and small subunit ribosomal DNA (ssu rDNA) PCR-RFLP patterns. The Mt DNA RFLP patterns were heterogeneous between species. The type strains of A. palestinensis and A. pustulosa showed almost identical patterns of isoenzymes and rDNA PCR-RFLP with an estimated sequence divergence of 2.6%. The other species showed heterogeneous patterns of isoenzymes and rDNA PCR-RFLP. It is likely that A. pustulosa is closely related with A. palestinensis and that the former may be regarded as a junior synonym of the latter.
Figures
Fig. 1 Microagraphs of Acanthamoeba cysts. A, A. culbertdoni; B, A. healyi; C, A. pustulosa; D, A. palestinensis; E, A. royreba; F, A. lenticulata. Bar = 10.0 µm.
Fig. 2 Polyacrylamide gel isoelectric focusing patterns for five kinds of isoenzymes of 6 strains of Acanthamoeba. AcP, acid phosphatase; ADH, alcohol dehydrogenase; GPI, glucose phosphate isomerase; LAP, leucine aminopeptidase; MDH, malate dehydrogenase. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. lenticulata; 6, A. royreba.
Fig. 3 Agarose gel electrophoretic restriction fragment patterns for mitochondrial DNA from 6 strains of Acanthamoeba. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. lenticulata; 6, A. royreba; M, Hind III digested lambda DNA for molecular size standard.
Fig. 4 Agarose gel electrophoretic restriction fragment patterns for PCR amplified ssu rDNA from 6 strains of Acanthamoeba. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. royreba; 6, A. lenticulata; M, DNA molecular size stanard.
Fig. 5 Phylogenetic tree of Acanthamoeba isolates based on genetic divergence estimates. The matrix of divergence estimates in Table 2 was used to construct this tree using UPGMA.
Tables
Table 1 Morphology and sources of Acanthamoeba strains analyzed in this study
Table 2 Proportions of homologous fragments and estimated average numbers of changes per nucleotide position
References
1.
Adam KMG. J Protozool 1964;11:423–430.
2.
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.
3.
Byers TJ, Hugo ER, Stewart VJ. Genes of Acanthamoeba: DNA, RNA and protein sequences (a review). J Protozool 1990;37(4):17S–25S.
4.
Chung DI, Kong HH, Yu HS, Oh YM, Yee ST, Lim YJ. Biochemical and molecular characterization of a strain KA/S2 of Acanthamoeba castellanii isolated from Korean soil. Korean J Parasitol 1996;34(1):79–85.
5.
Costas M, et al. Protistologica 1984;20:33–41.
6.
De Jonckheere JF. J Protozool 1983;30:701–706.
8.
Gast RJ, Byers TJ. Genus- and subgenus-specific oligonucleotide probes for Acanthamoeba. Mol Biochem Parasitol 1995;71(2):255–260.
9.
Gast RJ, Fuerst PA, Byers TJ. Discovery of group I introns in the nuclear small subunit ribosomal RNA genes of Acanthamoeba. Nucleic Acids Res 1994;22(4):592–596.
10.
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.
11.
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.
12.
Kong HH, Chung DI. 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 Parasitol 1996;34(2):127–134.
13.
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.
14.
Molet B, et al. Protistologica 1976;12:571–576.
15.
Moura H, Wallace S, Visvesvara GS. Acanthamoeba healyi n. sp. and the isoenzyme and immunoblot profiles of Acanthamoeba spp., groups 1 and 3. J Protozool 1992;39(5):573–583.
16.
Nei M, Li WH. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci U S A 1979;76(10):5269–5273.
17.
Page FC. Re-definition of the genus Acanthamoeba with descriptions of three species. J Protozool 1967;14(4):709–724.
18.
Pussard M, et al. Protistologica 1977;13:557–598.
19.
Reich K. Arch Protistenkd 1933;79:76–98.
20.
Singh BN, Das SR. Studies on pathogenic and non-pathogenic small free-living amoebae and the bearing of nuclear division on the classification of the order amoebida. Philos Trans R Soc Lond B Biol Sci 1970;259(832):435–476.
21.
Stratford MP, et al. J Gen Microbiol 1978;108:33–37.
22.
Visvesvara GS. Classification of Acanthamoeba. Rev Infect Dis 1991;13 Suppl 5:S369–S372.
23.
Willaert E, Stevens AR, Tyndall RL. Acanthamoeba royreba sp. n. from a human tumor cell culture. J Protozool 1978;25(1):1–14.
24.
Yagita K. Jpn J Parasitol 1993;42:468–478.
25.
Yagita K, Endo T. Restriction enzyme analysis of mitochondrial DNA of Acanthamoeba strains in Japan. J Protozool 1990;37(6):570–575.
PDF Links
Full text via DOI
Download Citation 
