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Preservation of wild isolates of human malaria parasites in wet ice and adaptation efficacy to in vitro culture.

Tantular IS, Pusarawati S, Khin L, Kanbe T, Kimura M, Kido Y, Kawamoto F - Trop Med Health (2012)

Bottom Line: These results indicated that a great difference in adaptation to in vitro culture may exist between wild isolates distributed in continental Southeast Asia and in eastern Indonesia and that gametocytogenesis might be easily switched on in Indonesian isolates.In wild isolates of P. vivax, P. malariae and P. ovale preserved for 2-9 days, ring forms or young trophozoites survived, but adaptation to in vitro culture failed.These results indicate that wild isolates can be preserved in wet ice for 9-10 days.

View Article: PubMed Central - PubMed

Affiliation: Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia ; Department of Parasitology, Faculty of Medicine, Airlangga University, Surabaya, Indonesia.

ABSTRACT
Wild isolates of malaria parasites were preserved in wet ice for 2-12 days and cultivated by a candle jar method. In four isolates of Plasmodium falciparum collected from Myanmar and preserved for 12 days, all failed to grow. In 31 isolates preserved for 5-10 days, nine were transformed to young gametocytes, but 22 isolates grew well. From Ranong, Thailand, nine isolates preserved for 7 days were examined, and six grew well. On the other hand, all of the 59 isolates collected from eastern Indonesian islands failed to establish as culture-adapted isolates, even most of them were preserved only for 2-3 days: 10 isolates stopped to grow, and 49 isolates were transformed to sexual stages by Day 10. These results indicated that a great difference in adaptation to in vitro culture may exist between wild isolates distributed in continental Southeast Asia and in eastern Indonesia and that gametocytogenesis might be easily switched on in Indonesian isolates. In wild isolates of P. vivax, P. malariae and P. ovale preserved for 2-9 days, ring forms or young trophozoites survived, but adaptation to in vitro culture failed. These results indicate that wild isolates can be preserved in wet ice for 9-10 days.

No MeSH data available.


Related in: MedlinePlus

Map showing the study sites.
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Related In: Results  -  Collection


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Figure 1: Map showing the study sites.

Mentions: Fresh isolates of the four human malaria parasites were obtained at several villages in Myanmar (Tanintharyi Division, Mandalay Division, Bago Division, Mon State, Rakhine State, Shan State), at a clinic in Ranong, Thailand, and at several villages in eastern Indonesian islands (Buru, Halmahera and Flores; Fig. 1) from March 1998 to December 2006 [4, 13–19]. Informed consent was obtained from all symptomatic volunteers before diagnosis. Thin and thick smears were made from a single finger puncture and examined using the acridine orange (AO) staining method [20, 21]. AO-stained thin smears were examined at a magnification of 400 using a light microscope and a halogen illuminator equipped with an interference filter for excitation of AO dye. If a malaria-positive patient was diagnosed as single-species infection with parasitaemia higher than 0.5%, informed consent was obtained again, and 2–3 ml of venous blood was drawn into an evacuated blood collection tube with sodium heparin powder. Confirmation of single infections with P. falciparum, P. vivax, P. malariae or P. ovale was performed later at laboratories in Indonesia and Japan by AO- and Giemsa-stained, thin smear microscopy and also by PCR-based diagnosis [16, 18].


Preservation of wild isolates of human malaria parasites in wet ice and adaptation efficacy to in vitro culture.

Tantular IS, Pusarawati S, Khin L, Kanbe T, Kimura M, Kido Y, Kawamoto F - Trop Med Health (2012)

Map showing the study sites.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3475313&req=5

Figure 1: Map showing the study sites.
Mentions: Fresh isolates of the four human malaria parasites were obtained at several villages in Myanmar (Tanintharyi Division, Mandalay Division, Bago Division, Mon State, Rakhine State, Shan State), at a clinic in Ranong, Thailand, and at several villages in eastern Indonesian islands (Buru, Halmahera and Flores; Fig. 1) from March 1998 to December 2006 [4, 13–19]. Informed consent was obtained from all symptomatic volunteers before diagnosis. Thin and thick smears were made from a single finger puncture and examined using the acridine orange (AO) staining method [20, 21]. AO-stained thin smears were examined at a magnification of 400 using a light microscope and a halogen illuminator equipped with an interference filter for excitation of AO dye. If a malaria-positive patient was diagnosed as single-species infection with parasitaemia higher than 0.5%, informed consent was obtained again, and 2–3 ml of venous blood was drawn into an evacuated blood collection tube with sodium heparin powder. Confirmation of single infections with P. falciparum, P. vivax, P. malariae or P. ovale was performed later at laboratories in Indonesia and Japan by AO- and Giemsa-stained, thin smear microscopy and also by PCR-based diagnosis [16, 18].

Bottom Line: These results indicated that a great difference in adaptation to in vitro culture may exist between wild isolates distributed in continental Southeast Asia and in eastern Indonesia and that gametocytogenesis might be easily switched on in Indonesian isolates.In wild isolates of P. vivax, P. malariae and P. ovale preserved for 2-9 days, ring forms or young trophozoites survived, but adaptation to in vitro culture failed.These results indicate that wild isolates can be preserved in wet ice for 9-10 days.

View Article: PubMed Central - PubMed

Affiliation: Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia ; Department of Parasitology, Faculty of Medicine, Airlangga University, Surabaya, Indonesia.

ABSTRACT
Wild isolates of malaria parasites were preserved in wet ice for 2-12 days and cultivated by a candle jar method. In four isolates of Plasmodium falciparum collected from Myanmar and preserved for 12 days, all failed to grow. In 31 isolates preserved for 5-10 days, nine were transformed to young gametocytes, but 22 isolates grew well. From Ranong, Thailand, nine isolates preserved for 7 days were examined, and six grew well. On the other hand, all of the 59 isolates collected from eastern Indonesian islands failed to establish as culture-adapted isolates, even most of them were preserved only for 2-3 days: 10 isolates stopped to grow, and 49 isolates were transformed to sexual stages by Day 10. These results indicated that a great difference in adaptation to in vitro culture may exist between wild isolates distributed in continental Southeast Asia and in eastern Indonesia and that gametocytogenesis might be easily switched on in Indonesian isolates. In wild isolates of P. vivax, P. malariae and P. ovale preserved for 2-9 days, ring forms or young trophozoites survived, but adaptation to in vitro culture failed. These results indicate that wild isolates can be preserved in wet ice for 9-10 days.

No MeSH data available.


Related in: MedlinePlus