Abstract
The effect of packaging, shipping temperatures and storage times on recovery of Bacillus anthracis. Sterne spores from swabs was investigated. Macrofoam swabs were pre-moistened, inoculated with Bacillus anthracis spores, and packaged in primary containment or secondary containment before storage at −15°C, 5°C, 21°C, or 35°C for 0–7 days. Swabs were processed according to validated Centers for Disease Control/Laboratory Response Network culture protocols, and the percent recovery relative to a reference sample (T0) was determined for each variable. No differences were observed in recovery between swabs held at −15° and 5°C, (p ≥ 0.23). These two temperatures provided significantly better recovery than swabs held at 21°C or 35°C (all 7 days pooled, p ≤ 0.04). The percent recovery at 5°C was not significantly different if processed on days 1, 2 or 4, but was significantly lower on day 7 (day 2 vs. 7, 5°C, 102, p=0.03). Secondary containment provided significantly better percent recovery than primary containment, regardless of storage time (5°C data, p ≤ 0.008). The integrity of environmental swab samples containing Bacillus anthracis spores shipped in secondary containment was maintained when stored at −15°C or 5°C and processed within 4 days to yield the optimum percent recovery of spores.
Highlights
During the Anthrax response in 2001, environmental surface samples were collected and shipped to laboratories for extraction and analysis [1,2,3]
We investigated the effects of storage conditions such as temperature, time, containment, and presence of dust on recovery of viable Bacillus anthracis (BA) spores from macrofoam swabs
No significant difference in recovery was seen between swabs pre-moistened with phosphate buffered saline with 0.02% Tween® 80 (PBST) and those pre-moistened with NB for all inoculum levels when the storage data up to 2 days for all temperatures was pooled (p=0.76)
Summary
During the Anthrax response in 2001, environmental surface samples (swabs, wipes, HEPA vacuum socks, etc.) were collected and shipped to laboratories for extraction and analysis [1,2,3]. Though Bacillus anthracis (BA) spores are known to be hardy, questions arose about the possibility that spores would germinate and allow for cell growth in the presence of dust and wetting agent. Another concern centered on temperature fluctuations that may occur during shipping and how this might impact the viability of spores before arriving at the laboratory [4]. Congressional reports from the Government Accountability Office [4,5] determined that answering these questions was critical in order to provide better confidence in the results of environmental sampling after an anthrax contamination event
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