In vitro culture of Plasmodium vivax liver stages underlies key understandings of the fundamental biology of this parasite, particularly the latent, hyponozoite stage, toward drug and vaccine development. Here, we report systematic production of Plasmodium vivax sporozoites in colonized Anopheles darlingi mosquitoes in the Peruvian Amazon. Human subject-derived P. vivax-infected blood was fed to Anopheles darlingi females using standard membrane feedings assays. Optimizing A. darlingi infection and sporozoite production included replacement of infected patient donor serum with naïve donor serum, comparing anticoagulants in processing blood samples, and addition of penicillin-streptomycin and ATP to infectious blood meals. Replacement of donor serum by naïve serum in the P. vivax donor blood increased oocysts in the mosquito midgut, and heparin, as anticoagulant, was associated with the highest sporozoite yields. Maintaining blood-fed mosquitoes on penicillin-streptomycin in sugar significantly extended mosquito survival which enabled greater sporozoite yield. In this study, we have shown that a robust P. vivax sporozoite production is feasible in a malaria-endemic setting where infected subjects and a stable A. darlingi colony are brought together, with optimized laboratory conditions.

中文翻译：

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从秘鲁亚马逊地区的定殖达氏按蚊连续供应间日疟原虫子孢子。

间日疟原虫肝阶段的体外培养是对该寄生虫的基本生物学（尤其是潜伏的次生子阶段）对药物和疫苗开发的基本了解的关键理解。在这里，我们报告了秘鲁亚马逊河中定殖的达氏按蚊的间日疟原虫子孢子的系统生产。使用标准的膜饲喂测定法，将人类受试者来源的间日疟原虫感染的血液饲喂给达氏按蚊雌性。优化达令吉氏菌的感染和子孢子的产生包括用纯净的供体血清代替被感染的患者供体血清，比较处理血样中的抗凝剂，以及在感染性血粉中添加青霉素-链霉素和ATP。间日疟原虫供体血液中的幼稚血清替代供体血清会增加蚊子中肠和肝素中的卵囊，作为抗凝剂，与子孢子的最高产量有关。在糖中的青霉素-链霉素上维持食血蚊子可以显着延长蚊子的生存时间，从而提高子孢子的产量。在这项研究中，我们已经表明，在疟疾流行的环境中，将感染的受试者和稳定的达林氏菌菌落聚在一起，并优化实验室条件，可以稳定地生产间日疟原虫子孢子。