PRACTICAL 1

INTRODUCTION
Chemicals are used either as antiseptics or disinfectants. Antiseptics are chemical agent that are
used typically i.e on the surface of living tissue to kill or inhibit the growth of microorganism.
Disinfectants are also used to kill microorganisms on inanimate objects. The chemicals used in
this practical are Dettol, 70% ethanol, hypo bleach and omo detergent, sterile distilled water was
used as the control test.

Dettol: it is used after surgery as antiseptic. The original Dettol liquid antiseptic
and disinfectant is light yellow in colour in the concentrated form but, as several
of the ingredients are insoluble in water, it produces a milky emulsion of oil
droplets when diluted with water, exhibiting the ouzo effect. The active ingredient
in Dettol that confers its antiseptic property is chloroxylenol (C8H9ClO), an
aromatic chemical compound. Chloroxylenol comprises 4.8% of Dettols total
admixture, with the rest made up by pine oil, isopropanol, castor oil, soap and

water[1].
Alcohols: alcohols, usually ethanol or isopropanol, are sometimes used as a
disinfectant, but moere often as an antiseptic (the distinction being that alcohol
tends to be used on living tissue rather than non living surfaces). They are noncorrosive, but can be a fire hazard. They also have limited residual activity due to
evaporation, which results in brief contact times unless the surface is submerged,
and have a limited activity in the presence of organic material. Alcohols are most
effective when combined with distilled water to facilitate diffusion through the
cell membrane; 100% alcohol typically denatures only external membrane
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proteins. A mixture of 70%ethanol or isopropanol diluted in water is effective

against a wide spectrum of bacteria, though higher concentrations are often

needed to disinfect wet surfaces.

Hypo bleach: hypo has become a synonymy of consumer bleach in Nigerian
household owing to its , affordability and accessibility. Hypos active ingredient is

sodium hypochlorite 3.5% w/v meant for household use[2].

Omo detergent: according to unilever, the active detergent components in OMO
powders sold in eurrope are surfactants. Such as sodium
dodecylbenzenesulfonaten. Some OMO powders also contain lipase, amylase and
other enzymes that help break down protein-based stains. Sodium compounds,
such as sodium carbonate and sodium sulfate, are common building and bulking
agents in these European formulations. These agents comprise most of the actual
mass of the powder. Other binding agents, structrants and stabilizers, sch as
cellulose, sodium polyacrylate and tetrasodium etidronate ensure that components

of the detergent remain well mixed[anonymous].

Sterile distilled water: this was used as control for the experiment.

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 70% ethanol was added to test
tube D 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
A
B
C
D
E

CHEMICAL AGENTS
Dettol
70% ethanol
Hypo bleach
OMO detergent
Sterile distilled water

NUMBER OF COLONIES
No growth
Growth all over the plate
No growth
No growth
27 colonies and smarmy
organisms

DISCUSSION
The growth on the 70% ethanol plate has proven that the ethanol used for the experiment is not
effective for the control of microorganism. This can either be due manufacturers defect or wrong
v/v ethanol to water ratio was used.
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.

CONCLUSION
In conclusion, it has been observed that chemical agents can be used in the control of
microorganisms(including the spore forming ones such as Bacillus subtillis)

APPENDIX FOR PRACTICAL 1

plate A: dettol

Plate D: omo detergent

Plate B: 70% ethanol

plate E: sterile distilled water

Plate C: hypo bleach

PRACTICAL 2
INTRODUCTION
Temperature, like pH, has an important effect on microbial growth. Based on response to a low,
medium or high temperature, microbes could even be grouped as psychrophiles, mesophiles or
thermophiles. However each microorganism has a range of temperatures that supports its growth
best. Below and above this range, growth may be adversely effected [3].
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
A
B
C

Equipment
Water bath
Oven
Refrigerator

Number of colonies
Growth all over the plate
54 colonies
Swarmy growth all over the

Freezer

plate
30 colonies, swarmy growth at

Work bench

the side of the pate

Swarmy growth all over the

plate

DISCUSSION
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 between the plates on the work bench to that of the water bath.

CONCLUSION
In conclusion, it has been noted from this experiment that temperature below 1000C is not
effective enough to kill spore forming bacteria such as Bacillus subtilis.

APPENDIX FOR PRACTICAL 2

Plate A: H202

plate c: refrigerator

plate B: oven

Plate D: freezer

Plate E: water bench

REFERENCES
1. Summary of product characteristic/Dettol Liquid. MHRA 22 November 2010. Retrieved
9 November 2014
2. www.hypo.com.ng/products.html
3. Fawole, M.O. and Oso, B.A. (2004). Laboratory manual of Microbiology, 5thEdition,
Spectrum

Books Ltd, Ibadan, Nigeria.

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