BI103 General Biology

PRACTICAL 3: Generation of Biogas by Indigenous Bacteria

(Archaebacteria) from Cow Dung
Objective:
To investigate the possibility of biogas production from a cheap raw material (cow dung) using a
laboratory scale digester.

Introduction:
Biogas is a mixture of colorless, flammable gases obtained by the anaerobic digestion of plant
based organic waste materials. Biogas is typically made up of methane (50-70%) carbon dioxide
(30-40%) and other trace gases. It is generally accepted that fuel consumption of a nation is an
index of its development and standard of living. There have been increases in the use of and
demand for fuel in terms of transportation and power generation in many nations including Fiji.
Fossil fuels are not environmentally friendly and are also expensive. The use of alternative and
more environmentally-friendly energy sources such as biogas has been advocated.

In Fiji, the use of wastes from organic matter, though important, has been relegated to the
background. There are abundant agricultural residues and municipal solid wastes, whose potentials
are yet to be fully tapped for energy generation. The possibility of using such wastes for biogas
production should be explored. The raw materials used in commercial methane generation include
plant residues, animal waste like cow dung and various urban wastes which are available in Fiji.
Biogas technology has advantages which include the following: generation of storable energy
sources, production of a stabilized residue that can be used as a fertilizer, an energy-efficient means
of manufacturing nitrogen containing fertilizer, a process having the potential for sterilization
which can reduce public health hazards from faecal pathogens, and if applied to agricultural
residues, a reduction in the transfer of fungal and plant pathogens from one year’s crop to the next.

Microorganisms are playing crucial role in the digestion of organic matter in nature. These
microbes are particularly methanogenic bacteria have an optimum pH around 7. The major source
of these organisms is rumen of animals. Biomethanogenesis takes place in three ways:
1. Solubilization and hydrolysis of the organic matter
2. Acidogenesis
3. Methanogenesis
In the first step, the conversion of complex organic substrates into butyric acid, propionic acid and
lactic acid. Secondly, the acetogenic bacteria convert those organic acids into acetic acid, hydrogen
and CO2. Then, the methanogenic bacteria reduce CO2 to methane and consume hydrogen. The
biogas produced by this process contain 65% methane, 30% CO2, 1% hydrogen and traces of N2,
O2, H2 and carbon monoxide (CO). The gas produced in trace amounts is collectively called as
marsh gas. It is colourless and burns with a blue flame. The energy produced by biogas 28 m3 is
equivalent to that produced by natural gas is 16.8 m3. The microbial isolates from the digesters
included Bacillus licheniformis, Escherichia coli and Clostridium sp., Bacillus sp., Yersinia sp.
and Pseudomonas species are responsible for biogas production in cow dung.

Material Required:

Cow dung, bottle, container, glass jars or burettes and trough.

Procedure:

The following procedures are done in laboratory to extract biogas from cow dung. It is stated for
your own information. For this lab, see the following video to understand the processes and
complete practical worksheet:
https://www.youtube.com/watch?v=MYj13FB_Iac
https://www.youtube.com/watch?v=sq-qNVhxZm0
https://www.youtube.com/watch?v=IGPEP9EZU3Y

Sample collection: Fresh cow dung was collected in a clean container with cover was used for
collection of the waste. If the cow dung was dried under the sun for four days and then pulverised
using a pestle and mortar. The pulverised dung was sieved and dried again for a day.

Slurry preparation: Three grams each of the fine powdered cow dung was weighed and mixed
with 30 cm3 of distilled water in a 250 ml conical flask or round bottom flask to give a ratio of
1:10. The mixture was thoroughly stirred with a glass rod to achieve homogeneity.

Anaerobic digestion: One set of 1 litre conical flasks or round bottom flasks, one flask was used
as digester. The flasks were labelled control and digester. A flask containing equal volumes of the
slurry (3 g dung: 10 cm3 water) was connected by a rubber delivery tube, which conveys the gas,
to a burette filled with water and placed in an inverted position in a glass trough containing water
such that gas released from the digestion process was collected in the burette by water displacement
method. The flask-end of the delivery tube was inserted into the mouth of the conical flask and
held in place by cotton wool stuffed at the mouth of the flask. The connecting point of tube and
flask was sealed with adhesive tape to prevent leakage of gas from the flask. The experimental set-
up was kept in the laboratory at room temperature (20 ± 2°C) with gas collection carried out over
water. The contents of the flasks were allowed to undergo digestion for a retention period of 3-4
weeks with weekly measurements of gas yields.

Results, Observations and Discussions:

Complete Part 1 and Part 2. Label, use arrows to show direction and explain the process of
biogas production. Watch the videos and do some research to answer the discussion questions.
Conclusion:

Write a brief conclusion.

Reference

USP library provides guide on how to reference using APA style. Use this link below to learn how to
do in-text citation and list references. https://usp.ac.fj.libguides.com/apa