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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
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Original Article

Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae

Hae-Jin Sohn1, Heekyoung Kang1, Ga-Eun Seo1, Jong-Hyun Kim2, Suk-Yul Jung3, Ho-Joon Shin1,*
The Korean Journal of Parasitology 2017;55(3):233-238.
Published online: June 30, 2017

1Department of Microbiology, Ajou University School of Medicine, and Department of Biomedical Science, Graduate School of Ajou University, Suwon 16499, Korea

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*Corresponding author (hjshin@ajou.ac.kr)
• Received: February 15, 2017   • Revised: April 4, 2017   • Accepted: April 10, 2017

Copyright © 2017 by The Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
Korean J Parasitol. 2017;55(3):233-238.   Published online June 30, 2017
DOI: https://doi.org/10.3347/kjp.2017.55.3.233
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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
Korean J Parasitol. 2017;55(3):233-238.   Published online June 30, 2017
DOI: https://doi.org/10.3347/kjp.2017.55.3.233
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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
Image Image Image Image Image
Fig. 1 Morphological changes of N. fowleri trophozoites into precysts (or cysts) cultured with encystation media for 6, 24, and 48 hr. Buffer 1 is an effective medium. Staurosporine induced rounding, but trophozoites were not recovered in Nelson’s medium.
Fig. 2 Morphological changes of A. castellanii trophozoites into precysts (or cysts) cultured in encystation media for 1, 2, and 3 days. The effective medium is buffer 2. Cysts recovered to trophozoites in PYG medium. Arrows indicate mature cysts.
Fig. 3 Morphological changes of A. polyphaga trophozoites into precysts (or cysts) cultured in encystation media for 1, 2, and 3 days. Buffer 2 is an effective medium. Cysts recovered to trophozoites in PYG medium.
Fig. 4 The mRNA expressions of nfa1 and nf-actin genes on N. fowleri trophozoites (Troph), precyts, and cysts (PreC) at 48 hr post cultivation with buffer 1. The p3 mRNA expression is used as the control.
Fig. 5 The mRNA expressions of actin and atg8 genes on A. castellanii (A) and A. polyphaga (B) trophozoites, precysts and cysts. The 18S rRNA mRNA expression is used as the control.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae

Compositions of encystation media for Naegleria (buffer 1), Acanthamoeba spp. (buffers 2 and 3), and Page’s amoeba saline

Buffer 1. Naegleria Encystment media (pH6.8)
 120 mM NaCI
 0.03 mM MgCI4
 1 mM Na2HPO4
 1 mM KH2PO4
 0.03 mM CaCI2
 0.02 mM FeCI2
Buffer 2. Acanthamoeba Encystment media (pH9.0)
 95 mM NaCI
 5 mM KCI
 8 mM MgSO4
 1 mM NaHCO3
 0.4 mM CaCI2
 20 mM Tris-CI (Ph9.0)
Buffer 3. Acanthamoeba Encystment media (pH7.6)
 0.1 mM KCI
 8 mM MgSO4
 0.4 mM CaCI2
 20 mM 2-amino-2-methlyl-1, 3-propanediol
Page’s amoeba saline (pH6.8–7.9)
 2 mM NaCI
 0.03 mM MgSO4
 0.03 mM CaCI2
 1 mM Na2HPO4
 1 mM KH2PO4
Table 1 Compositions of encystation media for Naegleria (buffer 1), Acanthamoeba spp. (buffers 2 and 3), and Page’s amoeba saline

Table 1

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