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1. To Study the Xenobiotic
Torent Bacteria
Bioremediation and Bioprocessing
ZOL-654 3(2-1)
BS 8th M (A & B)
Practical

2. Objective
 To understand how to isolate bacteria from mix culture
loaded with xenobiotics and characterize them after
isolation and confirmation

3. Theory
 Microorganisms play a significant role in
bioremediation of xenobiotics (pesticides), heavy metals
etc. which contaminated soil and wastewater.
 A xenobiotic is a chemical substance found within an
organism that is not naturally produced or expected to
be present within the organism.
 Xenobiotics may be grouped as carcinogens, drugs,
environmental pollutants, food additives,
hydrocarbons, and pesticides.
 Characterization of xenobiotic resistance in bacteria
provide a new clue for future perspectives reassert in
microbiology.

4. XenobioticTolerance
Tolerance means protection against something….
There are two major categories of protective
mechanisms:
(1) toxicokinetically derived mechanisms, which
alter the way in which organisms absorb,
biotransform, and excrete chemicals; and
(2) toxicodynamically derived mechanisms, in which
target sites are modified to reduce sensitivity.

5. Reasonnottogetsuccesstoisolatethe
resistantstrains
Three main reasons have been found.
 First, some compounds may be partly or completely degraded by
cooxidation or cometabolism that requires an additional substrate.
 Second, the compound in question may be degraded only by a
microbial consortium, with no single organism possessing all of the
required characteristics.
 Third, the degradation process may require interfaces and/or
gradients.
This last factor is applicable mainly to toxic and sparingly soluble
substances in aqueous environments.

6. Practical:
Isolationof dye degrading
bacteria

7. DYE
DEGRADING
BACTERIA

8. APPARATUS
 Pressure Cooker
 Stove
 Jam jars
 Beaker
 Petri plate
 Spreader and
 Syringe

9. MATERIAL
 Distilled water 100ml
 Agar Powder 0.3 g
 Nutrient Broth 0.4g
 Sample Dye

10. METHOD
1. Take 0.3 g nutrient broth powder in a glass jar.
2. Then add 100 ml distilled water in it and shake well.
3. Auto clave it for 15 minutes at 15 lb pressure and
then leave it at room temperature for about 15
minutes and let it cool.
4. Add 1 ml of textile effluent sample in the solution.
5. Observe the solution after 24 hours. As soon as the
whitish-cloudy growth appears we proceed to next
step.
6. For isolation of dye degrading bacteria spread 0.1m of
enrichment growth over the textile dye containing
nutrient agar plates.
7. Incubate it for 24 hours at room temperature and
then observe the bacterial colonies.

11. RESULT
 Bacterial growth appear on petri plate as different
shaped colonies
 Bacteria degraded about half of the total dye after 24
hours as indicated by the reduction in the intensity of
the dye colour in effluent .

12. Conclusion
 The present studies inform us that the isolated
bacteria have the properties to resist and accumulate
high levels of dye and pesticides; it may be harmful to
human being as well as to the animals.
 The isolation of these isolates comprise a valuable
assemblage for testing for strains degrading pollutants
in the presence of high concentrations of mercury,
which can be used in bioremediation of mixed wastes.

13. Resources
 https://www.researchgate.net/publication/278711261_X
enobiotic_ProtectionResistance_Mechanisms_in_Organ
isms
 https://link.springer.com/referenceworkentry/10.1007%
2F978-1-4419-0851-3_51
 https://aem.asm.org/content/aem/59/6/1717.full.pdf