ABSTRACT
The research study was aimed at investigating microorganisms associated with biogas production. Using sewage and cassava peel, as substrates. Sewage was collected from Ibrahim Babanginda hostel in Michael Okpara University of Agriculture, Umudike, Abia state while cassava peels were collected in Amawom, Umuahia, Abia state. Standard microbiological methods were used to isolate the organisms and anaerobic bio-digester was constructed to house the waste substrates for biogas production. Anaerobic bacteria isolated were identified as, Pseudomonas sp., Escherichia coli, Bacillus sp., Micrococcus sp., Proteus vulgaris, Citrobacter sp., Clostridium sp., Streptococcus bovis, Enterobacterium cloacae and Klebsiella sp., while fungi isolated were identified as Fusarium sp., Mucor sp., Aspergillus sp., Rhizopus sp. Methanogenic bacteria isolated were identified as Methanococcoides methyilutens, Methanoculleus bourgense, M. hungatei, Methanobacterium formicicum, M. hungatei, M. voltaei from media using anaerobic chamber for isolation . Analysis from this research revealed that the temperature of cassava peels and sewage ranged between 26°C and 30°C while the pH varied between 4.00 and 6.80 during digestion. The volume of biogas produced during the course of this project was between 1.6 and 8.2 ml and values for dissolve oxygen decrease from 10.4 to 0.0 mg/l. However, the study has shown that the role of Methanogens and other complementing bacteria and fungi in biogas production is indispensable.
TABLE OF CONTENTS
Title page
i
Certification ii
Dedication
iii
Acknowledgement
iv
Table of Contents
v
List of Tables
viii
List of Figures
ix
Abstract
x
CHAPTER ONE
1.0
Introduction 1
1.1 Aim and Objective
3
CHAPTER TWO
LITERATURE
REVIEW
2.0 Brief history
of biogas
4
2.1 Process/Principles of
biogas production
5
2.1.1
Hydrolysis
8
2.1.2 Acidogenesis (acidification phase)
8
2.1.3 Acetogenesis
9
2.1.4 Methanogenesis
10
2.2 Microorganisms
involved in biogas processes
11
2.2.1
Acidogens
11
2.2.2
Syntrophic Acetogens
12
2.2.3
Methanogens (Archaea)
13
2.3
Cassava peel and Sewage as substrates
for biogas production
16
2.3.1
Sewage
16
2.3.1.1
Types of Sewage
18
2.3.1.2
Reuse of Sewage
18
2.3.2
Cassava peels
19
2..3.3
Pretreatment of cassava peel and sewage for efficient
activity of microbial and biogas production 21
2.3.3.1 Pre-separation
21
2.3.3.2
Size reduction and disinfectant 21
2.4 Factors affecting
anaerobic degradation
22
2.4.1
Temperature 22
2.4.2
pH and Alkalinity
23
2.4.3
Oxygen 24
2.4.4
Ammonia Concentration
25
2.4.5
Hydraulic Retention Time and Organic Loading Rate
27
2.4.6
Inhibitor
28
2.4.7
Mixing
32
2.5
Importance of co-digestion
33
2.6
Types of digesters
33
2.6.1
Plug flow digesters
33
2.6.2
Fixed dome digesters
34
2.6.3
Floating drum digesters 35
CHAPTER
THREE
3.0
Materials and Methods
37
3.1 Biogas digester construction
37
3.2 Sample collection and preparation
38
3.3
Microbiological analysis
38
3.3.1 Isolation of microorganisms(Bacteria and
fungi)
38
3.3.2 Identification of bacterial and fungal
isolates
39
3.3.2.1 Morphological
and biochemical identification of bacteria and fungi isolates 39
3.5
Biochemical test 40-43
3.6
Analysis of physico-chemical parameters
43
3.7
Quantification of gas analysis
43
CHAPTER
FOUR
4.0 Results
44-51
CHAPTER
FIVE
5.0 Discussion
52
5.1
Conclusion
57
5.2
Recommendation
57
REFERENCES 58-67
LIST OF TABLES
Table
Title
Page
2.1:
Various
constituents of biogas generated from the anaerobic digestion process. 11
2.2:
Synthrophic
aceto-oxiding bacteria in association with hydrogentrophic methagens. 13
2.3: Inhibitors in anaerobic decomposition
processes and the concentrations at which
31
They become damaging.
4.1: Morphological characteristics of bacterial
isolates.
48
4.2:
Methane producing bacteria during the process of biogas production. 49
4.3:
Biochemical characterization and identification of bacterial
isolates 50
4.4:
Characterization and identification of fungal isolates 51
LIST OF FIGURES
Figure Title
Page
2.1: A schematic presentation of anaerobic
digestion process. 6
2.2: A schematic presentation of anaerobic
digestion process. 7
2.3:
Schematic representation of a plug flow digester 34
2.4: Schematic
representation of a fixed dome digester
35
2.5: Schematic
representation of a floating drum digester 36
3.1: Locally made biodigester
37
4.1: Shows pH variation of cassava peels and sewage during
fermentation process. 44
4.2: Shows the mesophilic temperature variation of
the substrates 45
(cassava peels and sewage)
before and during the digestion process.
4.3:
Shows Dissolve
oxygen decreasing from 10.4 - 0.0 during the digestion 46
process indicating consumption
of oxygen by facultative organisms.
4.4:
Shows the quantity of biogas produced during anaerobic digestion. 47
CHAPTER ONE
1.0 Introduction:
Millions of tons of wastes
(liquid or solid) are generated each year from municipal, industrial and
agricultural sources. Unmanaged organic waste fractions from farming, industry
and municipalities etc. decompose in the environment, resulting to large-scale
contamination of land, water and air, and also release green house gases. These
wastes not only represent a threat to environmental quality, but also possess a
potential energy value that is not fully utilized despite the fact that they
are cheap and abundant in most parts of the world (Merlin
Christy, et al., 2014). Biogas is one
of the versatile renewable fuels gotten or prepared from the organic wastes
which can be used for power and heat/cool production or it can be upgraded to
biomethane to be used as vehicle fuel (Bhuvaneswari et al., 2014).
Anaerobic
digestion is a suitable technology to treat organic wastes with aid of various
diversity of microorganism for biogas production. Anaerobic digestion consists
of several interdependent, complex sequential, and parallel biological
reactions, during which the products from one group of microorganisms serve as
the substrates for the next, resulting in transformation of organic matter
mainly into a mixture of methane and carbon dioxide with minor quantities of
nitrogen, hydrogen, ammonia and hydrogen sulfide etc (Ishmael et al., 2014; Merlin Christy et al., 2014). In nature, this process
occurs in environments such as hot springs, swamps, paddy fields, lakes and
oceans and the intestinal tract of animals like ruminants such as cattle etc.
Biogas is produced by anaerobic digestion of biological wastes such as cattle
dung, vegetable wastes, sheep and poultry droppings, municipal solid waste,
industrial waste water and land fill etc to give mainly methane (50-70%),
carbon dioxide (20-40%) and traces of other gases such as nitrogen, hydrogen,
ammonia, hydrogen sulphide, water vapour etc. these gases are smokeless,
hygienic and more convenient to use than other solid fuels (Gopinath et al., 2014).
In biogas production, the conversion of
complex organic matter to methane and carbon dioxide and other traces of gases
are possible mainly by the actions of different group of microorganisms, with
the microbial community of biogas comprised essentially of bacteria and fungi
and other groups of protozoan. The
essential microbial complex is comprised of
hydrolytic bacteria, fermenting bacteria, acetogenic bacteria and
methanogenic bacteria and these
groups of microorganisms have been
reported to establish synthrophic relationships where the later members of the
food chain depend on the previous for their substrate but may also have
significant metabolic products. These microorganisms act at the different
stages of the anaerobic process to bring about effective biogas production and
are an integral component of nature’s waste management and are commonly found
in soils and deep waters as well as land fill sites (Asikong et al., 2016).
The
anaerobic digestion is characterized by a series of biochemical transformations
of organic matter or wastes. The degradation steps are carried out by different
consortia of microorganisms, which partly stand in syntrophic interrelation and
place different requirements on the environment.
The
whole process involves several distinct stages such as hydrolysis, acidogenesis, acetogenesis and the final stage
methanogenesis. In stage 1, fats, complex carbohydrate and proteins are
hydrolysed to their monomeric form by enzymes. In stage 2, the monomers are further
degraded into short chain acids and these short chain acids are converted to
hydrogen, carbon dioxide and acetate and in the final stage which is stage 3,
the intermediate products are converted to methane and carbon dioxide by
methanogens (Ishmael et al., 2014).
1.1 AIM AND OBJECTIVE:
1.
To isolate and identify microorganisms from sewage and cassava peels
responsible for biogas production.
2.
To produce biogas from sewage and cassava peels.
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