CSI-1

Technical Data - MSDS - Test Data

                               

Efficacy of a Chemical Solutions International Product
Against Bacillus Anthracis Spores

Objective:
To determine the effect of a Chemical Solutions International (CSI) product on Bacillus anthracis spore viability. Spores will be treated with the product and then tested for the ability to germinate on culture medium.

Materials and Methods

Bacillus anthracis. Spores of strain 7702 were used for this study.

CSI Product. The product was supplied as 2%, 4%, and 6% solutions. The 2% and 6% solutions were used in this study.

Treatment Protocol.

  1. Add 0.9 ml of CSI solution to a 12 x 75 mm glass tube. For control tube, add 0.9 ml of water.
  2. Add 107 spores in 0.1 ml of water to each tube.
  3. Vortex briefly and incubate for desired time.
  4. Wash spores 3 times in 1% peptone as follows:
    Attach a Swinnex filter unit containing a sterile Millipore
    GS 0.22uM filter to the bottom of a 5-10 cc syringe.
    Using a glass pipet, transfer contents of a tube to the bottom of the syringe.
    Use the syringe plunger to force the liquid through the filter.
    Separate the filter unit from the syringe, remove the plunger, replace the filter unit, and add 5 ml of 1% peptone. Force the liquid through the filter as before.
    Repeat the washing 2 times.
  5. Carefully remove filter from filter unit and place it in a sterile 14-ml conical tube.
  6. Add 1 ml of 1% peptone to the conical tube and vortex to remove spores from the filter.
  7. Dilute in 1% peptone as indicated. Plate 0.1 ml aliquots on LB agar plates.
  8. Incubate at overnight at 37oC.
  9. Count colonies and determine count colony forming units (cfu) per ml.

 

 
2%
6%
Time (min) Expt 1
12/12		 Expt 2
12/17		 Expt 3
12/20		 Expt 1
12/17		 Expt 2
12/20
0 8.6 x 106 2.5 x 107 2.2 x 107 1.9 x 107 2.2 x 107
30 4.3 x 105 1.6 x 105 3.7 x 105 <10 4.8 x 102
60 1.6 x 103 2.3 x 102 3.3 x 103 <10 1.3 x 102
180 ND ND <10   <10


Conclusions

The data indicate that submersion of B. anthracis 7702 spores in the CSI solutions (2% and 6%) for 2 hrs results in complete loss of spore viability. Numbers of viable spores were reduced from approximately 107 to fewer than 10. In this assay 10 represented the limit of detection.

The rate of inactivation was greater for the 6% solution. The 6% solution reduced the number of viable spores from 107 to fewer than 500 after only 30 min of treatment (0.005% viability). The 2% solution reduced the number of viable spores from 107 to less than 5 x 105 after 30 min (5% viability).