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Simple and rapid method for detection of bacterial spores in powder useful for first responders.


* Bacillus anthracis is an aerobic, Gram-positive, spore-forming, nonmotile Bacillus species.

* Its spores germinate when they are in an environment rich in amino acids, nucleosides, and glucose, such as the blood or tissues of an animal or human host.

* Vegetative Bacillus species turn into spores in harsh environmental conditions, such as boiling, freezing, desiccation des·ic·ca·tion
n.
The process of being desiccated.



desic·ca
, and nutrient exhaustion.

* The spores can survive for decades in adverse conditions.

* Current detection methods, such as colony counting or polymerase chain reaction polymerase chain reaction (pŏl`ĭmərās') (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is  (PCR PCR polymerase chain reaction.

PCR
abbr.
polymerase chain reaction


Polymerase chain reaction (PCR) 
), require more than a working day to provide results.

* The need for a rapid method by which first responders can screen for the presence of spores was highlighted by the anthrax attack of 2001.

* The majority of powders that were sampled in the context of that attack were hoaxes.

* Hoaxes place a heavy burden on public health laboratories.

* A simple and rapid screening procedure that could detect spores in an unidentified powder would reduce the workload of the laboratories.

* Appropriate actions could be taken immediately upon detection of the spores.

* An adenosine triphosphate triphosphate /tri·phos·phate/ (tri-fos´fat) a salt containing three phosphate radicals.

tri·phos·phate
n.
A salt or ester containing three phosphate groups.
 (ATP ATP: see adenosine triphosphate.
ATP
 in full adenosine triphosphate

Organic compound, substrate in many enzyme-catalyzed reactions (see catalysis) in the cells of animals, plants, and microorganisms.
) bioluminescence bioluminescence (bī'ōl'mĭnĕs`əns), production of light by living organisms.  assay allows estimation of viable bacterial cells within minutes.

* Bacterial spores are, however, deficient in ATP.

* So they are almost undetectable by a regular bioluminescence technique.

* Therefore, they need to be germinated to a vegetative state by addition of a nutrient.

* The purpose of this study was to find the best conditions for triggering the breakdown of endospore en·do·spore
n.
1. A small spore formed within the vegetative cells of some bacteria.

2. A fungus spore borne within a cell or within the tubular end of a sporophore.

3. The inner layer of the wall of a spore.
 dormancy.

* The variables include germination time, temperature, nutrient type, and nutrient concentration.

* The optimum conditions for spore germination can be summarized as follows:

1. 37[degrees]C as a germination temperature,

2. 15 minutes as a germination time, and

3. single-strength solution of brain heart infusion as a nutrient.

* TSB TSB TPS (Thermal Protection System) Sample Box
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 can serve as a good alternative to brain heart infusion.

* Also, 5 minutes of germination time seems to be long enough to identify an increase in the biological activity of spores.

* The detection limit is less than 100 spores.

* All necessary equipment is portable.

* The germinants can easily be warmed with a small portable incubator or a hand warmer.

* Therefore, this method can be used to confirm the presence of viable spores in less time than traditional methods, at a higher level of sensitivity, and on site.
COPYRIGHT 2006 National Environmental Health Association
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Title Annotation:Practical Stuff!
Publication:Journal of Environmental Health
Geographic Code:1USA
Date:Apr 1, 2006
Words:367
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