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

最近，在切萨皮克疫情爆发前的近十年里，在美国大陆第二大河口Albemarle-Pamlico的几条支流爆发的有毒疫情中，被称为有毒Pfiester Complex的恐鞭毛虫对鱼类健康和人类健康的潜在微妙但严重的慢性/亚致命性影响已经意识到。在这项研究中，我们将在定制设计的生物危害III设施中大规模生产鱼腥藻的有毒培养物，用于毒性阶段的研究。这一设施将用于维持Pfiester piscicida和第二个已知的有毒类Pfiestia物种的毒性阶段，并用于确定每一物种有毒阶段之间阶段转化的范围的所有实验。这将涉及从Neuse和Pocomoke河口每隔8至12个月克隆一次有毒和无毒的Pfiester piscicida分离株，以及第二个已知的有毒类Pfiester属物种(需要克隆，因为与活鱼在有毒模式下维持较长时间的培养往往会失去毒性)。这项研究将极大地提高对鱼类死亡/家畜现场样本中Pfiester的识别，并将加强对有毒Pfiester复合体中这两个已知物种的阶段转化和丰度的营养控制的一般了解。

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.