Bacterial endosymbiosis within the cytoplasm of <i>Acanthamoeba lugdunensis</i> isolated from a contact lens storage case

, Chung, Dong Il; , Kong, Hyun Hee; , Kim, Tae Ho; , Hwang, Mee Yul; , Yu, Hak Sun; , Yun, Ho Cheol; , Seol, Sung Yong

doi:1997.35.2.127

Korean J Parasito > Volume 35(2):1997 > Article

Brief Communication


Korean J Parasitol. 1997 Jun;35(2):127-133. Korean. Published online Jun 20, 1997. http://dx.doi.org/10.3347/kjp.1997.35.2.127
Copyright © 1997 by The Korean Society for Parasitology


Bacterial endosymbiosis within the cytoplasm of Acanthamoeba lugdunensis isolated from a contact lens storage case
Dong-Il Chung,^*¹Hyun-Hee Kong,¹Tae-Ho Kim,¹Mee-Yul Hwang,¹Hak-Sun Yu,¹Ho-Cheol Yun,¹ and Sung-Yong Seol²
¹Department of Parasitology, Kyungpook National University School of Medicine, Taegu 137-701, Korea.
^*Corresponding author (Email: dichung@bh.kyung)

Received January 10, 1997; Accepted May 12, 1997.
Abstract
Transmission electron microscopy of an Acanthamoeba isolate (KA/L5) from a contact lens case revealed bacterial endosymbionts within cytoplasm of the amoebae. The Acanthamoeba isolate belonged to the morphological group II. Based on the polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) of 18S ribosomal RNA coding DNA (rDNA), the isolate was identified as A. lugdunensis. Strain typing by isoenzyme analysis using isoelectric focusing (IEF) and mitochondrial (Mt) DNA RFLP revealed that the isolate was closely related with KA/L1, the most predominant type of isolates from contact lens storage cases, KA/E2, a clinical isolate, KA/W4, previously reported to host endosymbionts, and L3a strains of A. lugdunensis. The endosymbionts were similar to those of KA/W4 in aspects that they were randomly distributed in both trophozoites and cysts, and were rod-shaped bacteria measuring approximately 1.38 x 0.50 µm. But the number of endosymbionts per amoeba was significantly lower than that of KA/W4. They were neither limited by phagosomal membranes nor included in lacunaelike structure.

Figures


	Fig. 1 Agarose gel electrophoretic of PCR products and digested DNA fragments of Acanthamoeba KA/L5 and reference strains. Lane 1:Acanthamoeba KA/L5, 2:Acanthamoeba KA/L1, 3:Acanthamoeba lugdunensis L3a, M:Hind III digested lamda phage DNA, A: Amplisize^R (Biorad, USA) as DNA molecular size standard.


	Fig. 2 Acrylamide gel electrophoretic patterns of digested PCR product of Acanthamoeba KA/L5 and reference strains. Lane 1:Acanthamoeba KA/L5, 2:Acanthamoeba KA/L1, 3:Acanthamoeba lugdunensis L3a, M: Amplisize^R (Biorad, USA) as DNA molecular size standard.


	Fig. 3 Zymograms for isoenzymes of KA/L5 and reference strains of A. lugdunensis separated by polyacrylamide gel isoelectric focusing in pH gradient 3-10. AcP, acid phosphatase; ADH, alcohol dehydrogenase; LAP, leucine aminopeptidase; GPI, glucose phosphate isomerase. Lane 1:A. lugdunensis KA/L5, 2: KA/L1, 3: KA/W4, 4: KA/W2, 5: KA/E2, 6: L3a strain.


	Fig. 4 Agarose gel electrophoretic patterns of digested mitochondrial DNA of A. lugdunensis KA/L5 and reference strains. Lane 1:A. lugdunensis KA/L5, 2: KA/L1, 3: KA/W4, 4: KA/W2, 5: KA/E2, 6: L3a strain, M:Hind III digested lamda phage DNA as DNA molecular size standard. KA/L5 and KA/W4 show extra bands (arrows) as well as common bands with the other reference strains.


	Fig. 5 Electron micrographs of A. lugdunensis KA/L5 with bacterial endosymbionts. A: Rod-shaped bacterial endosymbionts are randomly distributed in the cytoplasm of trophozoite. B: Cyst with endosymbionts. C: Magnification of A. Ribosomes of amoebic host are studed on the surface of endosymbionts. D: Dividing endosymbiont. Bar indicates 2 µm.


	Fig. 6 Agarose gel electrophoresis of PCR products with Legionella specific primer set. Chromosomal DNA of endosymbionts and Legionella spp. were used as template DNA. Lane 1: endosymbionts of Acanthamoeba KA/L5 strain, 2:L. pneumophila ATCC 33154, 3:L. pneumophila ATCC 33156, 4:L. gormanii ATCC 33297, M: Amplisize as DNA molecular size standard.

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