| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
Gametocytogenesis in Leucocytozoon caulleryi was investigated in in vitro culture using erythrocytes infected with the second-generation merozoites. In 1992, cultures conditions such as several kinds of media and air phases, and effects of addition of human normal crythrocytes were examined. The influences on ages of chicken, infection doses and days of infection were also examined. As a result, when cultures were carried out at 37? C in an atomosphere of 2% O_2,8% CO_2 and 90% N_2, the second-generation merozoites on Day 15 after infection can be developed into mature gametocytes in culture using improved RPMI 1640 medium supplemented with 25mM Hepes, 23.8mM sodium bicarbonate, 2.1mM L-glutamine, 0.37mN hypoxanthine, and addition of horse serum (10%) and human normal erythrocytes. However, the number of mature gametocytes observed were very smaller than that of in vivo. In 1993, the concentration of medium and NaCl, and addition of vitamines and heparin were examined in culture. The l
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arger number of developed gametocytes were observed in culture using slightly high concentration of medium or NaCl. The supernatant culture of anti-Plasmodium falciparum antibody producing hybridoma cells, and Berenil of anti-protozoa drug, which have gametocytogenesis activity in cultured P falciparum, were also examined, but these reagents have not gametocytogenesis activity in cultured L,caulleryi. In 1994, the efficacy of artificial bleeding from infected chickens were examined in vivo and in vitro. After artificial bleeding on Day 12, the number of immature erythrocytes in peripheral blood increased and reached a first peak on Day 14. In this time, most of the second-generation merozoites invaded into immature erythrocytes, so that the larger number of mature or developed gametocytes were observed in infected chickens after artificial bleeding or in vitro culture. However, the phenomenon that the second-generation merozoitesinvade into immature erythrocytes was not considered to be under the necessary and sufficient conditions for doveloping from the second-generation merozoites into mature gametocytes Less
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