Abstract
Spores of Bacillus anthracis Sterne strain were recovered from 100 ml and 1 L volumes of tap and source waters using filtration through a 0.45 um filter, followed by overnight culture on agar plates. In a set of experiments comparing sheep red blood cell (SRBC) plates with a chromogenic agar formulation designed by R & F Laboratories, with a spiking dose of 47 plate-enumerated spores in 100 ml tap water, the mean spore recoveries were 34.0 and 30.8 spores, respectively. When a spiking dose of 100 fluorescence activated cell sorter(FACS)-enumerated spores was used in 100 ml potable water, the average recovery with SRBC plates was 48 spores. Detection efforts with spiking doses of 35 and 10 spores in 1 L tap water were successful, but recovery efforts from spiked 1 L volumes of source water were problematic due to the concomitant growth of normal spore-forming flora. Recoveries were also attempted on 10 L volumes of tap water. For a spiking dose of 100 spores, mean recovery from six replicates was 11 spores (±6.8, range 2–20), and for a spiking dose of 10 spores, mean recovery from six replicates was 2.3 spores (±3.5, range 0–9). Efforts were also made to “direct detect” spores via polymerase chain reaction (PCR) on washes from filters. When spiking 534 spores in 100 ml, 9/9 replicates of spiked tap water, 6/6 source water replicates, and 0/3 unspiked controls were positive by lef PCR. When 534 spores were spiked into 1 L tap water, the lef PCR was unsuccessful; however, using the nested vrrA PCR resulted in 4/9 spiked samples, and 0/3 unspiked controls, testing positive. Our results indicate that an inexpensive and user-friendly method, utilizing filtration apparatus commonly present in many water quality testing labs, can readily be adapted for use in detecting this potential threat agent.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.