Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16

Yamasaki, Masahiro; Hwang, Shiang-Jyi; Ohta, Hiroshi; Yamato, Osamu; Maede, Yoshimitsu; Takiguchi, Mitsuyoshi

doi:https://doi.org/10.14943/jjvr.55.4.129

Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16

Yamasaki, Masahiro; Hwang, Shiang-Jyi; Ohta, Hiroshi et al.

Japanese Journal of Veterinary Research, 2008, 55(4), 129-136

Permalink : https://doi.org/10.14943/jjvr.55.4.129

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アクセス権	open access
DOI	https://doi.org/10.14943/jjvr.55.4.129
URI	https://hdl.handle.net/2115/32387
タイトル	Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16
著者	Yamasaki, Masahiro ; Hwang, Shiang-Jyi ; Ohta, Hiroshi ; Yamato, Osamu ; Maede, Yoshimitsu ; Takiguchi, Mitsuyoshi
e-Rad_Researcher 40322846 著者名 Yamasaki, Masahiro
著者名 Hwang, Shiang-Jyi
e-Rad_Researcher 50431333 著者名 Ohta, Hiroshi
著者名 Yamato, Osamu
e-Rad_Researcher 40002084 著者名 Maede, Yoshimitsu
e-Rad_Researcher 70261336 著者名 Takiguchi, Mitsuyoshi
言語	英語
キーワード	Babesia gibsoni
	flow cytometry
	SYTO16
発行日	2008-03-07
出版者	The Graduate School of Veterinary Medicine, Hokkaido University
収録物名	Japanese Journal of Veterinary Research
巻(号)	55(4)
ページ	129-136
記述	In the present study, we employed flow cytometry to evaluate the level of parasitemia of Babesia gibsoni infecting canine erythrocytes in vivo and in vitro by using fluorescent nucleic acid staining. Peripheral blood samples from a B. gibsoni-infected dog and cultured B. gibsoni parasitizing in canine erythrocytes were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16. In this study, we utilized normal canine erythrocytes (LK erythrocytes) and canine erythrocytes containing high concentrations of potassium, reduced glutathione, and some free amino acids (HK erythrocytes) as host cells for culture. Parasitized cells in vivo were discriminated completely from unparasitized cells and a correlation (r = 0.998) between the percentage of SYTO16-positive cells and parasitemia in vivo was observed. On the other hand, erythrocytes in vitro could not be divided clearly into parasitized and unparasitized cells. However, when LK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost the same as, and was well correlated (r = 0.932) with, the level of parasitemia. When HK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost half of, but was correlated (r = 0.982) with, the level of parasitemia. Therefore, we attempted to observe the changes in the percentage of parasitized cells after treatment with antiprotozoal drug or mitochondria inhibitors by using flow cytometry. The changes in the percentage of SYTO16-positive cells corresponded well with the changes of the level of parasitemia when the parasites in HK erythrocytes were cultured with each compound. The present results suggest that flow cytometric detection using SYTO16 is a rapid and reliable method for monitoring parasitemia both in vivo and in vitro.
資源タイプ	紀要論文
出版タイプ	Version of Record
PISSN	0047-1917