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

最近，在切萨皮克爆发前近十年，在美国大陆最大的河口Albemarle-Pamlico的几个支流中，被称为有毒Pfiesteria复合体的鞭毛藻对鱼类健康和人类健康的潜在微妙但严重的慢性/亚致命影响被意识到。在这项研究中，我们将在一个专门设计的生物危害III设施中大规模生产piscicida有毒培养物，用于毒性阶段的研究。该设施将用于维持鱼腥味Pfiesteria Pfiesteria的毒性阶段和第二种已知的有毒Pfiesteria样物种，并用于所有实验以确定每个物种的毒性阶段之间的阶段转化范围。这将涉及每隔8至12个月从Neuse和Pocomoke河口克隆有毒和无毒的Pfiesteria Pfiesteria，以及第二种已知有毒的Pfiesteria样物种（这是必要的，因为与活鱼长期保持有毒模式的培养往往会失去毒力）。这项研究将极大地提高对鱼类死亡/动物流行病现场样品中Pfiesteria的认识，并将提高对有毒Pfiesteria复群中两种已知物种的阶段转化和丰度的营养控制的一般理解。

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.