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Microbe Inotech Laboratories, Inc. has developed several tests for determining if and how well bioremediation can occur at your site. 1. Endpoint Assay This analysis will indicate if any of the indigenous, or purchased strains of bacteria we have isolated are able to use your chosen contaminant or substrate as a carbon source. It also reveals how well the bacteria can use the carbon source compared to hundreds of other strains that have been tested. The Endpoint Assay uses the 96 well microtiter plate to give rapid testing with increased savings. In this test, the microplate is loaded with a suspension of your isolated bacteria, the contaminant you have chosen, and media and water for the positive and negative controls. Each unknown and control is done in replicates of at least four wells for added accuracy. After 24 hours incubation the plate is read in a microplate reader to determine the optical density of each of the wells. A dye in the wells turns purple if the bacteria are using the carbon source. The higher the optical density, the better your strains are at consuming the contaminant. The data is reported to you in an easy to read bar chart (see right).
2. Kinetics Study  Once
you have determined there are degraders present, and you would like to know how
the addition of nutrients or oxygen will affect their growth rate, you may want
to do a kinetics study. The Kinetics
Study is
similar to the Endpoint
Assay in that it uses the 96 well microtiter plate, containing a
tetrazolium dye that darkens proportionately to the metabolic activity
of the
strain. The kinetic plate is put into the microplate reader immediately
after
preparation. The optical density of the wells is then read every 10
minutes for
18 hours. Each reading is automatically plotted to create a growth
curve.
Comparisons of curves from wells with nutrients to the control wells
gives your precise information about how the addition of any;
substrates will affect the growth of your strains.
3. Comparative Population Assay This is a MPN (most probable number) test that gives you the order of magnitude of your degrader populations. The 96 well plate is also used in this test. The Comparative Population Assay differs from the Endpoint or Kinetic Assays since a serial dilution of your soil or water sample is used directly in the plate, along with the contaminant, instead of the isolated strains. After 48 hours incubation, the plate is read in the microplate reader to determine the highest dilution at which growth is detected. This gives you the MPN of the degrader populations. Also in the plate are positive growth control wells. These wells provide you with the total heterotrophic plate count information. |
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