A field investigation of Bacillus anthracis contamination of U.S. Department of Agriculture and other Washington,D.C., buildings during the anthrax attack of October 2001

In response to a bioterrorism attack in the Washington, D.C., area in October 2001, a mobile laboratory (ML) was set up in the city to conduct rapid molecular tests on environmental samples for the presence of Bacillus anthracis spores and to route samples for further culture analysis. The ML contained class I laminar-flow hoods, a portable autoclave, two portable real-time PCR devices (Ruggedized Advanced Pathogen Identification Device [RAPID]), and miscellaneous supplies and equipment to process samples. Envelopes and swab and air samples collected from 30 locations in the metropolitan area once every three days were subjected to visual examination and DNA extraction, followed by real-time PCR using freeze-dried, fluorescent-probe-based reagents. Surface swabs and air samples were also cultured for B. anthracis at the National Veterinary Service Laboratory (NVSL) in Ames, Iowa. From 24 October 2001 to 15 September 2002, 2,092 pieces of mail were examined, 405 real-time PCR assays were performed (comprising 4,639 samples), and at the NVSL 6,275 samples were subjected to over 18,000 platings. None of the PCR assays on DNA extracted from swab and air samples were positive, but viable spores were cultured from surface swabs taken from six locations in the metropolitan area in October, November, and December 2001 and February, March, and May 2002. DNA extracted from these suspected B. anthracis colonies was positive by real-time and conventional PCRs for the lethal factor, pXO1, and for capA and vrr genes; sequence analysis of the latter amplicons indicated >99% homology with the Ames, vollum, B6273-93, C93022281, and W-21 strains of B. anthracis, suggesting they arose from cross-contamination during the attack through the mail. The RAPID-based PCR analysis provided fast confirmation of suspect colonies from an overnight incubation on agar plates.     

Temperature sensitivity of soil respiration in a low-latitude forest ecosystem varies by season and habitat but is unaffected by experimental warming

Experimental warming of forest ecosystems typically stimulates soil respiration (CO₂ efflux), but most warming experiments have been conducted in northern latitudes (>?40°N) with relatively young soils. We quantified the influence of experimental warming on soil respiration (R_T) in two adjacent forest habitats—a mature, closed canopy forest and a gap where trees were manually removed— on highly-weathered Ultisols of the southeastern U.S. (33°N). Using temperature variation, both natural and induced by experimental warming, we also quantified the temperature sensitivity of R_T, defined as the activation energy, E_A in the Arrhenius equation. Experimental warming (either + 3 °C or + 5 °C above ambient) did not significantly increase soil respiration rate or cumulative CO₂ loss over the 3 years of the experiment, and did not influence the temperature sensitivity of soil respiration, once the influence of natural temperature variation was taken into consideration. Despite the absence of an experimental warming effect, we observed that E_A varied on monthly time scales, and varied differently in each habitat. Soil moisture and habitat also influenced R_T, but the effects were not consistent, and varied by month. Our results suggest that although R_T does depend on temperature, the sensitivity of R_T to temperature variation is influenced primarily by factors like microclimate and plant phenology that can change on relatively short (<?monthly) time scales. Thus, using the temperature sensitivity of R_T to predict future CO₂ losses due to warming is only reasonable if monthly variation in E_A is incorporated into models for lower-latitude subtropical ecosystems with highly weathered soils, such as those in this study. Finally, our results suggest that higher temperatures may not enhance R_T in highly-weathered, C-poor soils to the extent that has been reported in prior studies of high-latitude soils, which may constrain ecosystem-atmosphere carbon exchanges and feedbacks to the climate system.