Production of Toxic Culture of Pfiesteria piscicida in a Specialized Biohazard III Facility

最近，在切萨皮克爆发之前的近十年里，美国大陆最大河口的几个支流在第二大河口Albemarle-Pamlico爆发有毒物质期间，人们认识到了被称为有毒Pfiesteria复合体的dino-flagellates对鱼类健康和人类健康的微妙但严重的慢性/亚致死影响。 在本研究中，我们将大量生产的有毒培养的杀鱼原甲在定制设计的生物危害III设施的毒性阶段的研究。 该设施将需要用于维持杀鱼Pfiesteria和第二种已知的有毒类Pfiesteria的毒性阶段，以及用于所有实验以确定每个物种的毒性阶段之间的阶段转换范围。 这将涉及以8至12个月的间隔从纽斯河口和波科莫克河口克隆有毒和无毒的杀鱼性Pfiesteria piscicida分离株，以及第二种已知有毒的类Pfiesteria物种（需要，因为与活鱼保持毒性模式较长时间的培养物往往会失去毒性）。 这项研究将大大提高识别Pfiesteria现场样品从鱼杀死/流行病，也将提高营养控制阶段的转换和丰富的两个已知物种的有毒Pfiesteria复杂的一般理解。

Recently, the potential for subtle but serious chronic/sublethal impacts from dino-flagellates known as the toxic Pfiesteria complex on fish health as well as human health was realized during toxic outbreaks in several tributaries of the largest estuary on the U.S. mainland, in the second largest estuary, the Albemarle-Pamlico, for nearly a decade before the Chesapeake outbreaks. In this research we will mass-produce toxic culture of P. piscicida in a custom-designed biohazard III facility for research on toxic stages. This facility will be required for maintenance of toxic stages of Pfiesteria piscicida and the second known toxic Pfiesteria-like species, and for all experiments to determine the range of stage transformations among toxic stages of each species. This will involve cloning toxic and nontoxic isolates of Pfiesteria piscicida, and of the second known toxic Pfiesteria-like species, from the Neuse and Pocomoke Estuaries at 8- to 12-month intervals (needed since cultures that are maintained in toxic mode with live fish for longer periods tend to lose virulence). This research will greatly improve recognition of Pfiesteria in field samples from fish kills/epizootics, and will also enhance general understanding of trophic controls on stage transformations and abundance of both known species in the toxic Pfiesteria complex.