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Volume 55, Number 4 >

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

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/jjvr.55.4.129

Title: Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16
Authors: Yamasaki, Masahiro Browse this author →KAKEN DB
Hwang, Shiang-Jyi Browse this author
Ohta, Hiroshi Browse this author →KAKEN DB
Yamato, Osamu Browse this author
Maede, Yoshimitsu Browse this author →KAKEN DB
Takiguchi, Mitsuyoshi Browse this author →KAKEN DB
Keywords: Babesia gibsoni
flow cytometry
SYTO16
Issue Date: 7-Mar-2008
Publisher: The Graduate School of Veterinary Medicine, Hokkaido University
Journal Title: Japanese Journal of Veterinary Research
Volume: 55
Issue: 4
Start Page: 129
End Page: 136
Description: 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.
Type: bulletin (article)
URI: http://hdl.handle.net/2115/32387
Appears in Collections:Japanese Journal of Veterinary Research > Volume 55, Number 4

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