BADMUS, SIMBIAT

ATINUKE
10/48KC031
MCB 424 PRACTICAL
REPORT
INTRODUCTION

Microorganisms are everywhere and will almost always interfere with some processes in our
daily activities. It's required that they are "absent" in some environments before production can
proceed otherwise, they would contaminate products meant for use and may be a source of harm
to man and his general environments. An example of such is the pharmaceutical industries where
chemotherapeutic agents for human use are produced. It is essential that this products are of
themselves "clean" of "other sources of harm" before they can be used.

The destruction or, removal of microorganisms from instruments, materials, body surfaces, et.c
is referred to as decontamination. Decontamination involves the use of chemical and physical
agents. Chemical agents such as alcohol, heavy metals, et.c are used to achieve this process.
Examples of physical agents include heat, low temperature, radiation, et.c.

The effectiveness of these agents against microorganisms depend on the type of microorganism
and the stage in the life cycle at which they are exposed to the agents. It is because of this, that it
is essential to understand the type of organism that contaminate and its relative resistance in
order to determine the ideal method of decontamination.

Practical 1

Title: The evaluation of chemical agents of microbial control on some bacteria

Aim: to evaluate chemical agents on microbial control on some bacteria

Materials used

Broth culture of Bacillus subtilis

Chemical agents labeled as follows
A. Dettol
B. 70% ethanol
C. Hypo bleach
D. Omo detergent
E. Sterile distilled water
5 plates of nutrient agar
5 sterile cotton bulbs
10 sterile test tubes
10ml sterile distilled water
Procedure

Work bench and our hands were sterilized with 70% ethanol. All the practical work was done
beside the flame in other aseptic conditions to be maintained. The sterile test tubes and nutrient
agar plate were labeled; A, B, C, D and E. 5ml of each of the organism was transferred into each
of the test tubes. 1ml of Dettol was added to test tube A;1ml of 1ml of hypo bleach was added to
test tube C ;1g of omo detergent was added to test tube D; 1ml of 70% ethanol was added to test
tube B and 1ml of sterile distilled was added to test tube E using separate syringes. The test
tubes were shook gently to mix the contents. The test tubes were placed in test tube rack and
allowed to stand for 10minutes. 0.1ml was inoculated into labeled nutrients agar plates from the
corresponding test tube sterile syringe. The inoculums were spread using sterile cotton bulb. All
plates were incubated at 37oC for 18hrs.The plates were removed from the incubator and
counted.

RESULT

PLATES CHEMICAL AGENTS NUMBER OF COLONIES

A Dettol No growth
B 70% ethanol Growth all over the plate
C Hypo bleach No growth
D OMO detergent No growth
E Sterile distilled water 27 colonies and smarmy
organisms

Conclusion

The growth on the 70% ethanol plate has proven that the ethanol is not effective.

The growth on the distilled water plate proves that the organisms used for the experiment are
viable (not dead)

The lack of growth on the other plates has proven the effectiveness of the chemical agent in the
control of microbial growth.

PRACTICAL 2

Title: Evaluation of physical techniques of microbial control

Aim: To evaluate the physical techniques in microbial control

MATERIALS USED

Broth culture of Bacillus subtilis

Oven (set at 100oC),
 water bath (set at 100oC),
Refrigerator (set at 15oC),
freezer take the temperature(-2oC)
work bench (28oC)
5 plates of nutrients agar
10 sterile syringe (1ml)
5 tubes of sterile peptone/saline water (5ml)
5 sterile cotton bulbs

PROCEDURE

The test tubes containing sterile peptone water were labeled A, B, C, D and E. 1ml of the
organism was transferred into each of the test tubes. Test tube A was placed in the water bath, B
in the oven, C in the refrigerator, D in the freezer, E on the work bench; they were all incubated
for 30minutes. After 30 minutes, the test tubes were removed and left on the bench to bring to
room temperature. 0.1ml was transferred into the tubes onto the center of nutrient Agar medium
and spread on the agar medium using sterile cotton bulb. All plates were incubated at 37oC for
18hours. After incubation the number of colonies on each plate was counted.

RESULT

Plates Equipment Number of colonies

A Water bath Growth all over the plate
B Oven 54 colonies
C Refrigerator Swarmy growth all over the
plate
D Freezer 30 colonies, swarmy growth at
the side of the pate
E Work bench Swarmy growth all over the
plate

Conclusion

In conclusion,

A. the water bath allows the growth of microorganisms all over the plate
B. it was observed the oven temperature which was set at 100oc was effective against the
microbial cells but not against the spores which causes the little growth on the plate.
C. The refrigeration only altered the growth for a short period of time but does not reduce
the microbial population
 D. The freezer reduced the number of organisms compared to that of the refrigeration
probably because some of the microbial cells were killed.
E. There is no difference on plates on the work bench to the water bath.

APPENDIX FOR PRACTICAL 1

plate A: dettol Plate B: 70% ethanol Plate C: hypo bleach

Plate D: omo detergent plate E: sterile distilled water

APPENDIX FOR PRACTICAL 2

Plate A: H202 plate B: oven

 plate c: refrigerator Plate D: freezer

Plate E: water bench