FACULTY : ENGINEERING

TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY

EDITION:
REVISION NO:

FACULTY OF ENGINEERING TECHNOLOGY

EFFECTIVE DATE:
EXPERIMENT: BACTERIA
DEPARTMENT
OF
CIVIL
ENGINEERING
TECHNOLOGY
COUNT
AMENDMENT DATE:

2
FEB
2015

ENVIRONMENTAL ENGINEERING TECHNOLOGY

LABORATORY
LABORATORY INSTRUCTION SHEETS
COURSE CODE

BNP 20503

EXPERIMENT CODE

EXPERIMENT 5

EXPERIMENT TITLE

BACTERIA COUNT

DATE
GROUP NO.
LECTURER/INSTRUCTOR/TUTOR

1)
2)

DATE OF REPORT SUBMISSION

DISTRIBUTION OF MARKS FOR

LABORATORY REPORT

EXAMINER COMMENTS:

ATTENDANCE/PARTICIPATION/DISIPLINE:

/5%

INTRODUCTION:

/5%

PROCEDURE:

/5%

RESULTS & CALCULATIONS

/15%

ANALYSIS

/15%

DISCUSSIONS:

/20%

ADDITIONAL QUESTIONS

/15%

CONCLUSION

/10%

SUGGESTIONS & RECOMENDATIONS

/5%

REFERENCES:

/5%

TOTAL:

/100%

RECEIVED DATE AND STAMP:

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BACTERIA
COUNT

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB
2015

AMENDMENT DATE:

STUDENT CODE OF ETHICS

DEPARTMENT OF CIVIL ENGINEERING TECHNOLOGY

FACULTY OF ENGINEERING TECHNOLOGY

I hereby declare that I have prepared this report with my own efforts. I also admit to
not accept or provide any assistance in preparing this report and anything that is in it
is true.
1) Group Leader
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________

2) Group Member 1
Name
:
Matrix No :

__________________________________________(Signature)
__________________________________
___________________________________

3) Group Member 2
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________

4) Group Member 3
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________
2

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BACTERIA
COUNT

1.0

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB
2015

AMENDMENT DATE:

OBJECTIVES
a)

To determine the total count of bacteria quantity in water sample by

performing total plate count.

2.0

LEARNING OUTCOMES
At the end of this course students are able to:
a) Be more proficient at dilutions.
b) Be more proficient at performing a standard plate count and determining
bacterial counts in a sample.

3.0

INTRODUCTION
3.1 The standard plate count method consists of diluting a sample with
phosphate buffer diluent until the bacteria are diluted enough to be
counted accurately. Hence, the final plates in the series should have
between 30 and 300 colonies. Fewer than 30 colonies are not acceptable
for statistical reasons (too few may not be representative of the sample),
and more than 300 colonies on a plate are likely to produce colonies too
close to each other to be distinguished as distinct colony-forming units
(CFUs).
3.2 The assumption is that each viable bacterial cell is separate from all others
and will develop into a single discrete colony (CFU). Thus, the number of
colonies should give the number of bacteria that can grow under the
incubation conditions employed. A wide series of dilutions (e.g., 10-4 to
10-10) is normally plated because the exact number of bacteria is usually
unknown.
3.3 Greater accuracy is achieved by plating duplicates or triplicates of each
dilution. Increased turbidity in a culture is another index of bacterial
growth and cell numbers (biomass). By using a spectrophotometer, the
amount of transmitted light decreases as the cell population increases.
The transmitted light is converted to electrical energy, and this is indicated
on a galvanometer. The reading, called absorbance or optical density,
indirectly reflects the number of bacteria. This method is faster than the

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BACTERIA
COUNT

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB
2015

AMENDMENT DATE:

standard plate count but it has limitation where sensitivity is restricted to

bacterial suspensions of 107 cells or greater.
4.0

INSTRUMENTS /APPARATUS / CHEMICAL / REAGENTS

4.1

Chemicals / Reagents
No chemical / chemical required in this experiment

4.2.

Apparatus /Instruments
i)

Petri plate

ii)

Pipette

iii)

Test tube

iv)

Glass rod

v)

Bunsen burner

vi)

Incubator

vii)

Ethanol 95% @ methanol

viii) Sterilizer
ix)

Microscope

x)

Bacteria medium:

Peptone = 5g, Beef Extract=3g, Agar=15g,

Distilled water=600 mL
5

PROCEDURE
5.1 Media preparation
Please prepare the nutrient media using the microbiology
standard method.
5.2 Sample preparation
Prepare the serial dilution of the water sample using the
appropriate dilution factor.
5.3 Plating procedures
Using the pour plate and spread plate method.

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BACTERIA
COUNT

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB
2015

AMENDMENT DATE:

RESULTS CALCULATIONS
6.1

Results
Table 6.1: Plating method

Plating
Method

Average
Colony
/plate

Dilution
1/10

1/100

1/1000

1/104

1/105

1/106

Total
bacteria
/ mL

Pour
Plate
Spread
Plate

ANALYSIS
Please show the calculation for each of the plating method and fill in the above
table. Analyze the results by using appropriate method. Explain your findings.

DISCUSSIONS
State the systematic bias error that could occur during this experiment.
Usually, the results give different readings for both methods. However, in
some cases, both methods produce the same results. Explain why the results
are indistinguishable.

ADVANCED QUESTIONS
1. Explain the meaning of a phrase two times ten to the eight cells per mL
in your own convenient terminology.
2. What the meaning of TNTC and the significant amount due to the TNTC?
Give the formula for determining bacteria count.
3. Design an experiment to compare the bacteria counts in different water
samples (tapwater, lake water, swimming pool water and rainbarrel water).
Explain the difference of bacteria count for each type of water sample?
4. In many experiments there are 2 types of control used which are positive
and negative control. Based on this experiment what is the suitable
control? How will the control affect your findings?

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BACTERIA
COUNT

10

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB
2015

AMENDMENT DATE:

CONCLUSION
Conclusion is merely a summary, presented in a logical order, of the
important findings already reported in the discussion section. It also relates to
the objectives stated earlier.

Prepared by/Disediakan oleh :

Approved by/Disahkan oleh :

Signature/Tandatangan :
Name/Nama : DR. NOR HASLINA HASHIM

Signature/Tandatangan :
Name/Nama : DR. SURAYA HANI ADNAN

Date/Tarikh :

FEBRUARY 2015

Date/ Date/Tarikh : FEBRUARY 2015