DEVELOPMENT OF AN AXENIC CULTURE FOR PNEUMOCYSTIS

描述：（改编自申请人的摘要）垫博士最近 成功地建立了一个无菌培养系统， Pc的生长，类似于组织培养物的生长。 是 本项目的总体目标是无菌培养Pc.第一特定 目的是试图确定产生 最健康的生物体进行培养 为加快文化发展， 研究人员还计划开发快速评估方法， 微生物数量和活力。 这些方法将应用于 完成第二个目标，无菌培养Pc.研究者 将尝试开发一种支持Pc生长的培养基， 在营养评估和生化分析之后， 研究人员已经成功地为其他人建立了这样的文化， 原生生物 各种浓度的营养块通常需要 在本发明中，将评估微生物培养的Pc生长。 无菌培养基或改进的粗培养基。 每种的最佳浓度 将通过缩小稀释范围来识别组织块。 一旦 确定，块的最佳浓度将保持恒定 而另一个块被测试和优化。 增长和活力将是 通过多种方法进行评估，包括放射性标记的前体 掺入、微生物计数和荧光测定。 第三 该项目的目标是试图描述生命周期阶段的特征， 无菌培养中的生长动力学。 超微结构和动力学 分析将用于确定生命周期的各个阶段， 培养，确定生长速度，并监测健康的 有机体 由于寄生原生生物通常会降低它们的毒性， 当长期培养时， 与表面抗原相连，培养生物体的抗原性 将进行超微结构评价。 这些项目旨在 首先，提供评估生物体数量的新的或改进的方法， 生存能力 第二，预计关于 从受感染的肺中分离出的生物体的生理状态将是 得到了 最后，希望通过加强对 Pc的代谢能力，此外， 将获得Pc的培养物。

DESCRIPTION: (Adapted from applicant's abstract) Dr. Cushion has recently succeeded in establishing an axenic culture system which supports limited growth of Pc, similar to that achieved with tissue cultures. It is the overall goal of this project to axenically culture Pc. The first specific aim is to attempt to identify isolation procedures that produce the healthiest organisms for culture. To expedite culture development, the investigator also plans to develop rapid methods for evaluation of organism numbers and viability. These methods will then be applied to accomplish the second goal, to axenically culture Pc. The investigator will attempt to develop a medium which supports the growth of Pc by following nutritional evaluations and biochemical analyses used by other investigators who have successfully established such cultures for other protists. Various concentrations of blocks of nutrients commonly required for culture of microbes will be evaluated for Pc growth in the present axenic medium or an improved crude medium. Optimal concentrations of each block will be identified by narrowing the range of dilutions. Once identified, the optimal concentration of a block will remain constant while another block is tested and optimized. Growth and viability will be assessed by a variety of methods including radiolabeled precursor incorporation, enumeration of organisms and fluorometric assay. The third goal of the project is to try to characterize the life cycle stages and kinetics of growth in axenic culture. Ultrastructural and kinetic analyses will be used to identify the life cycle stages replicating in cultures, determine the growth rate, and monitor the health of the organisms. Since parasitic protists commonly decrease their virulence when cultured over long periods and this virulence has oftentimes been linked to surface antigens, the antigenicity of the cultured organisms will be ultrastructurally evaluated. The projects have been designed to provide, first, new or improved methods of assessing organism number and viability. Secondly, it is anticipated that information on the physiologic state of organisms isolated from infected lungs will be obtained. Lastly, it is hoped that increased understanding of the metabolic capabilities of Pc and, in addition, the development of axenic culture for Pc, will be obtained.