Cultivation in vitro of Plasmodium gallinaceum oocysts |
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| Authors: | I Schneider |
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| Affiliation: | 1. Evolutionary Studies Institute, University of the Witwatersrand, Johannesburg 2050, South Africa;2. Karoo Palaeontology, Iziko South African Museum, Cape Town 8000, South Africa;3. Karoo Palaeontology Department, National Museum, Bloemfontein 9300, South Africa;4. University of the Free State, Department of Zoology and Entomology, Bloemfontein 9300, South Africa;5. Negaunee Integrative Research Center, Field Museum of Natural History, 1400 South DuSable Lake Shore Drive, Chicago, IL 60605, United States;1. Department of Anthropology, University of Hawai''i at Manoa, 2424 Maile Way 346 Saunders Hall, Honolulu, HI 96822, USA;2. Key Laboratory of Vertebrate Evolution and Human Origins of Chinese Academy of Sciences, Institute of Vertebrate Palaeontology and Palaeoanthropology, Chinese Academy of Sciences, No. 142 Xizhimenwai Street, Beijing 100044, China;3. Key Laboratory of Geobiology and Environmental Geology, Ministry of Education, China University of Geosciences, Wuhan, Hubei 430074, China;4. School of Earth Sciences, China University of Geosciences, Wuhan, Hubei 430074, China;5. Guangxi Museum of Nationalities, Nanning 530022, China |
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| Abstract: | Large numbers of viable infectious Plasmodium gallinaceum sporozoites, virtually free of mosquito tissue, were obtained by culturing somewhat younger stages of the sporogonous cycle. Twenty or more oocysts were placed in each hanging- or sitting-drop culture. Grace's insect tissue culture medium, slightly modified and supplemented with either vertebrate and/or invertebrate sera, was employed. In addition, approximately one half of the cultures contained mosquito cells, organs, or whole body extracts. Nine-day-old oocysts usually completed development within 24 hours and liberated sporozoites shortly thereafter. The extent of development in 8-day-old oocysts was dependent upon whether sporoblast formation with subsequent budding of the sporozoites had or had not occurred prior to explantation. If the former, the oocysts continued their development in 60% of the cultures; if the latter, less than 1% of the oocysts were capable of producing sporozoites. Attempts to culture 6- and 7-day-old oocysts were not successful. This failure of the younger oocysts to develop in vitro was attributed to insufficiencies in the culture medium, lack of tissue bulk, suboptimal conditioning of the medium, and the relatively static environment of the present system. The suggestion was made that such factors might be largely negated if the oocysts were cultured in the presence of an actively growing mosquito cell strain. |
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