There is a critical time between exposure and the onset of symptoms during a biological attack, where early detection and warning could save many lives. Over the past 15 years, members of the Environmental Biology Group, along with colleagues across LLNL and federal agencies, have been developing and fielding laboratory diagnostics to scan the environment to detect and respond to a biological attack as early as possible. Our Biological Aerosol Sentry and Information System (BASIS) deployable laboratory, which incorporated highly-specific DNA signatures and real-time Polymerase Chain Reaction (PCR) assays was deployed shortly after the events of 9/11 to protect the National Capital Region (NCR). We have continued to expand and improve upon methods, assays, and laboratories for environmental biothreat monitoring, which have been utilized in programs such as the Department of Homeland Security (DHS) BioWatch Program. We’ve supported the DHS BioWatch Program since it started in 2003 by providing subject matter expertise, reach-back support, data management systems, hands-on training, and even setting up and operating laboratories. Our current research is focusing on improving and developing methods for characterizing the public health risk of environmental contamination following an initial biothreat detection.

Dr. Carlsen, along with scientists from the Environmental Restoration Department (ERD), is investigating the ecological impact of uranium contamination around a former firing table at the Building 812 Complex at Site 300. The Building 812 Complex was built in the late 1950s and early 1960s to conduct explosive tests and diagnostics in support of national defense programs. ERD began investigating the area in 1988, and is currently focusing on characterizing and developing remedial alternatives for elevated uranium concentrations found in surface soil surrounding the Building 812 Complex, as well as in the underlying ground water. As the area supports a rich, natural ecosystem, Dr. Carlsen is investigating the movement of uranium through the ecosystem through the uptake by plants and invertebrates, and modeling the potential impact of uranium to higher vertebrates, specifically the deer mouse and rock wren. Preliminary results show that plants and invertebrates have taken up uranium, but the ecological significance of this uptake has not yet been determined.

Team members partnered with EPA’s National Homeland Security Research Center to develop rapid methods for analysis of viable pathogen cells or spores in surface and environmental samples. The RV-PCR method was successfully transitioned to the EPA Environmental Response Laboratory Network allowing EPA to be in a better position to make response and remediation decisions. After a bio-attack, a large number of samples of diverse type will need to be analyzed in a short time-frame to determine the efficacy of decontamination and to clear the site for re-use and re-entry. Routine methods including real-time PCR do not provide information on whether the decontamination efforts have killed the pathogen. The RV-PCR method is an innovative integration of high-throughput sample processing, short-incubation broth culture, and highly sensitive and specific real-time PCR assays to detect low levels of viable pathogen cells or spores. This RV-PCR method addresses limitations of the traditional culture method, and has a lower limit of detection (LOD) since the entire sample is used for analysis. The method is also semi-automated, and less prone to inhibition by environmental matrices and to interference by outgrowth of other microbes.

Dr. Carlsen, along with scientists from the Environmental Functional Area, is investigating the use of fire to manage A. grandiflora populations at Site 300. Earlier research at the original Site 300 A. grandiflora experimental population by Dr. Carlsen (Carlsen et al., 2000) showed A. grandiflora to prefer growing in a matrix of native perennial bunch grasses, as opposed to the exotic annual grasses that are now more common in California. As fire is a common tool used to increase the density of native perennial bunch grasses, the use of fire to provide a more hospitable habitat for A. grandiflora is being investigated. A long-term fire frequency experiment was initiated in 2000. A. grandiflora and P. secunda (a native perennial bunch grass common at Site 300) was established in twenty experimental plots. Plots were assigned to treatments that included a control (no burning), low frequency burning (every 5 years), intermediate frequency burning (every 3 years), and high frequency burning (every year). The final burn treatment was conducted in 2011. Three years of post-treatment data are being collected. Preliminary results suggest that while intermediate and high frequency burns do increase the density of native perennial bunch grasses, A. grandiflora does not tolerate the immediate impacts of the burn.