Experiment No: 1

Name of Experiment: Effect Of PH on Microbial Growth

Principle:
Different microorganisms often require distinct environments, with varied temperature, levels of
oxygen, light and acidity or pH level. Some microbes grow faster in environments with
extremely low pH values. These are called acidophiles, because of their preference for acidic
environments. Although most microorganisms requires neutral pH values to have optimum
growth, alkaliphilic microorganisms prefer low-acidity or high pH environment.

Acidophiles

Microorganisms which optimum growth at pH levels lower than 5 are called acidophiles. These
microbes are found in a variety of environments, including geysers and sulfuric pools, as well as
in the human stomach. Examples of acidophiles include the microscopic algae Cyanidium
caldarium and Dunaliella acidophila. The microscopic fungi, Acontium cylatium,
Cephalosporium and Trichosporon cerebriae can grow near pH 0. A primitive microorganism
called Picrophilaceae have optimum pH values close to zero and can also grow at negative pH
values.

Alkaliphilic

Alkaliphilic microorganisms have optimum growth at pH values between 9 and 12. These
microorganisms thrive in alkaline lakes, soils and other high pH environments. In slag dumps of
Lake Calumet, southeast Chicago, the water can reach a pH of 12.8, which is similar to caustic
soda. Some bacteria related to the Clostridium and Bacillus live in that extremely alkaline
environment. Mono Lake, in California, and Octopus Spring in Yellowstone Park are examples
of environments were alkaliphilic microorganisms are found.

Neutrophiles

Neutral pH values, laying between 6 and 8, are more commonly found in nature. Along their
evolution, most microorganisms have adapted to have optimum growths in acidity-neutral
environments. These microorganisms are called neutrophiles, and include most species of
microalgae and other organisms that form the phytoplankton, as well as some soil-dwelling
bacteria and yeasts.

Pathogens and PH

Most microorganisms associated with human, animals and plant diseases are neutrophiles, such
as Escherichia coli, which causes intestinal infections; Erwinia caratovora, a plant parasite;
Pseudomonas aeruginosa, which causes a series of infection in humans and animals; and
Streptococcus pneumoniae, which causes pneumonia. However, pathogens are also found among
acidophiles and alkaliphilics. The bacterium Lactobacillus acidophilus have optimum growth in
low pH levels, causing vaginal infections. The alkaliphilic bacterium Vibrio cholerae causes
cholera in humans.

Materials:
1.Trypton Soya Broth
2.Organism B
3.Distiled water

Equipments:
1.Loop
2.Bunsen Burner
3.Spectrophotometre
4. Cuvette

Procedure:

1. Inoculate organism B from a Petridis to the Tripton Soya broth

2. Incubate for 48 hours at 37C
3. Fill a blank Cuvette with the broth media
4. Select a cuvette filled with distilled water and place it in the spectrophotometer. Close the
lid.
5. Click on 0 ABS 100%T button, the instrument now reads 0.00000 A.

6. Choose a solution with known concentration and measure the absorbance between the
wavelengths 660 nm

7. Record the wavelength at the maximum absorbance value.

Observation: after 48 hours incubation at 37C

Result:
Absorbance value for the Organisms are given below

Organism pH Absorbance value Remark

A 4 0.049 low absorbance, low growth
B 10.5 0.017 Low absorbance, Low growth
C 6 0.192 High absorbance, High growth
D 9 0.037 Low absorbance, Low growth

Discussion:
The purpose of this experiment was to observe bacterial growth under different pH. For this
purpose we used Tripton Soya broth and after incubating we use spectrophotometer to check the
turbidity by measuring the absorbance. The higher the absorbance is ,the higher is the growth.
According to the organism A the absorbance was 0.049 for the pH 4. It can be consider as low
absorbance, that means low growth and this is due to the acidic pH. Organism B and D both were
incubate in alkaline pH, they were respectively 10.5 and 9. Their absorbance was respectively
0.017 and 0.037. Showing that the organism D had more growth than the organism B. Which had
higher pH. So pH 9 was more favorable for the growth. In case of organism C ,the pH was 6
which was among the optimum pH so the absorbance was high,0.192 indicating a high microbial
growth. According to the experiment it is shown that microorganism from well in neutral pH
rather than very high or low pH.