ANNEXURE A

FINAL%

FACULTY OF …
DEPARTMENT OF …
ASSIGNMENT COVER SHEET
MODULE TITLE Prokaryote classification and microbial technique
MODULE CODE 4MCB211
ASSIGNMENT TOPIC Turbidity measurement

LECTURER NAME DR F TSHABUSE

DUE DATE 29/04/2024
NON - PLAGIARISM DECLARATION
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wrong. I understand that my research must be accurately referenced. I have followed the rules and conventions
concerning referencing, citation and the use of quotations as set out in the Departmental Guide. This assignment is my
own work, or my group’s own unique group assignment. I acknowledge that copying someone else’s assignment, or part
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acceptable similarity percentage, I may receive 0 (ZERO) for my assignment.
LEARNER NAME LEARNER NO SIGNATURE
Mosisidi N 202273891
Mbelu S.N 202313283
Magwaza N.S 202344227
Magubane S.A 202320704
Kheswa A.Z 202339472
Nene P 202213709
Gumede S.L 202278362
Majola K 230001365
LECTURER REMARKS

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Introduction
Serial dilutions are used in microbiology to decrease the bacterial population to a
required concentration. Turbidity measurement is the technique used to indirectly
measure the concentration of bacterial population by measuring the cloudiness or
haziness of the broth culture which is caused by the presence of microorganisms
(bacteria). Turbidity measurement can be used to estimate the cell density of the
microorganisms in the broth culture, which is essential for various applications, such
as antibiotic susceptibility testing, microbial enumeration, and fermentation processes.
It can also be used to assess the effectiveness of antimicrobial agents or other
inhibitors on microbial growth. By measuring turbidity, you can gain valuable insights
into microbial growth and behavior, enabling informed decisions in various fields, such
as microbiology, biotechnology, and food safety. However, for determinations on
extracts (e.g. antibiotics and vitamins) of bacteria the turbidimetry method is no longer
useful and it is necessary to resort to weighing or chemical determination. In 1907,
MacFarlane introduced the first standard turbidimetry preparation for use in
bacteriology. Brown studied and extended the system. Both MacFarlane and Brown
referred the results of the opacity determination to the number of bacilli measured by
use of counting chambers. Their turbidimetric observations were performed with the
naked eye. In this report we are going to measure and discuss the effect of light
dispersed by colloidal suspension in a solution.

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Aim: To estimate the amount of cells in a suspension of bacteria by
turbidity measurement

Material / Apparatus
 Clean cuvettes
 2 3ml sterile pipettes
 1 spectrophotometer
 5 test tubes
 nutrient broth culture

Procedure
1. Prepare your broth cultures by inoculating nutrient broth with the desired microorganism.

3. Label five test tubes and add 9 ml of nutrient broth to each.

4. Using a sterile pipette, transfer 1 ml of each culture to the corresponding test tube,
ensuring each test tube contains the same volume of culture.

5. Mix the contents of each test tube thoroughly by gently swirling.

6. Measure the initial turbidity of each culture using a spectrophotometer at 620 nm

wavelength.

7. Record the initial optical density (OD) readings for each test tube.

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 Turbidity Measurement of Broth Culture

Results

OD Readings of Dilutions

Dilutions OD 620nm

0 (Original) 1,36

1:10 0,221

1:100 0,034

1:1000 0,013

1:10,000 0,009

1:100,000 0,006

Blank

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 OD VS DILUTION OF BACTERIAL SAMPLE

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Discussion of Results

The results plotted on a linear graph makes it clear that from test tube A to test tube E
the bacterial population is decreasing, thus serial dilution is performed successfully.
Turbidity is directly proportional to the concentration of bacterial cells present in the
medium (), as result, the decrease in bacterial population means the turbidity is also
decreasing respectively. The optical density (measure of the absorbance) is greater in
quantity in test tube A and decreases respectively towards test tube E, proving that
the absorbance of light is parallel to the concentration of bacterial population. These
results are in coherent with Lambert-beer’s law which states that the absorbance of
light is directly proportional to both the concentration of the absorbing medium and the
thickness of the medium.

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Conclusion
In conclusion, the turbidity measurement experiment conducted on the broth culture
effectively demonstrated the growth and density of microorganisms present in the
culture over time. The results obtained indicate a clear correlation between optical
density readings and microbial growth, with an increase in turbidity directly
corresponding to an increase in cell concentration. This relationship is in line with the
principles of turbidimetric measurements, where the scattering of light by particles in
suspension is used to quantify microbial growth (Madigan et al., 2018).

Furthermore, the consistency and reproducibility of the turbidity measurements

highlight the reliability of this method for monitoring microbial growth in liquid cultures.
By utilizing turbidity as a proxy for cell density, researchers can efficiently track the
growth kinetics of microorganisms without the need for time-consuming and labor-
intensive cell counting techniques (Madigan et al., 2018).

Overall, the turbidity measurement experiment provided valuable insights into the
dynamics of microbial growth in the broth culture, underscoring the utility of
turbidimetric methods in microbiology research and bioprocess monitoring.

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References
Madigan, M. T., Martinko, J. M., Bender, K. S., Buckley, D. H., & Stahl, D. A. (2018). Brock
Biology of Microorganisms (15th ed.). Pearson.
McFarland J (1907). Nephelometer: An instrument for media used for estimating the number
of bacteria in suspensions used for calculating the opsonic index and for vaccines
J. Spaun (1962) Bull. Wld Hlth Org., 26, 225

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