ISOLATION AND CHARACTERIZATION OF MICROORGANISMS INVOLVED IN BIOGAS PRODUCTION USING CASSA PEELS

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Product Code: 00008892

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Abstract

This study was aimed at investigating microorganisms associated with biogas production using cassava peel as substrates. Cassava peels obtained after peeling cassava roots were anaerobically digested using a 5 litre capacity fermentor. The peels were blended and mixed with water, in the ratio of 1:2. Standard microbiological methods and anaerobic biodigester were used to screen the isolates and the wastes substrate for biogas production. The mean flammable biogas yield of the cassava peels was 13.7Litre/total mass of slurry. Analysis revealed that the temperature of raw substrates ranged between 26°C and 30°C while the pH varied between 4.04 and 6.86 during digestion. The results of the total heterotrophic and anaerobic microbial counts during the digestion period showed a steady variation in the anaerobic bacteria and fungal counts as fermentation progressed. The load obtained for the total heterotrophic bacteria and anaerobic bacteria were within 2.9x104cfu/ml to 9.4x104cfu/ml and 1.8x103cfu/ml to 6.9x103cfu/ml respectively. Similarly the fungal count obtained through the digestion period ranged from 1.1x10cfu/ml to 3.9x10cfu/ml. The bacterial isolates were identified as Pseudomonas sp, Escherichia coliBacillus sp, Clostridium sp, and Methanococcus sp, while fungi isolated were identified as Saccharomyces cerevicaeAspergillus sp and Penicillium sp.





TABLE OF CONTENT

 

Title page                                                                                                                                i

Certification                                                                                                                           ii

Declaration                                                                                                                             iii

Dedication                                                                                                                              iv

Acknowledgement                                                                                                                  v

Table of content                                                                                                                      vi

List of Table                                                                                                                           vii

List of Figures                                                                                                                         viii

Abstract                                                                                                                                  ix


CHAPTER 1:

INTRODUCTION

1.1      Introduction                                                                                                            1

1.2    Aim and Objectives                                                                                                       4


CHAPTER 2:

LITERATURE REVIEW

2.0      Literature Review                                                                                                       5

2.1.      Biogas                                                                                                                         5

2.2       Biogas Composition                                                                                                   5

2.3       Chemical Characteristics of Biogas                                                                           6

2.4       Physical Characteristics of Biogas                                                                             6

2.5       Methane as a Component of Biogas                                                                           6

2.6       Collection of Biogas                                                                                                   7

2.7       Types of Biogas bioreactor                                                                                        7

2.8       Substrate  (Cassava Peels )                                                                                         7

2.9       Microbial Floral Present in Production of Biogas                                                      8

2.10     Microbiology and Biochemistry of Biogas Production                                              8

2.10.1  Hydrolysis                                                                                                                   10

2.10.2 Acidification/Acetogenesis                                                                                        10

2.10.3 Methane Formation                                                                                                    10

2.11     Interactions between the Various Microbial Groups                                                  11

2.12     Factors Influencing Biogas Production.                                                                     11

2.12.1  Temperature                                                                                                               11

2.12.2  pH (Hydrogen Concentration)                                                                                   12

2.12.3  Substrate Quality and Characterization                                                                     13

2.12.4  Loading Rate                                                                                                             13

2.12.5    Bioreactor  Design                                                                                                    14

2.13     Benefit of Biogas Technology                                                                                    14

2.14     Domestic and Industrial Uses                                                                                     15

2.15    Other Uses of Biogas                                                                                                   15


CHAPTER 3:

3.0       MATERIALS AND METHODS

3.1       Construction of Anaerobic Digester                                                                           16

3.2       Sample Collection                                                                                                      16

3.3       Pretreatment of Cassava Peels                                                                                    16

3.4       Charging of the Digester                                                                                            16

3.5       Determination of Quantity of Biogas Produced                                                         16

3.6       Determination of Physiochemical Analysis of the Slurry in the Bioreactor.                  17

3.6.1                Hydrogen Ion Concentration (pH)                                                                  17

3.6.2                Temperature                                                                                                   17

3.6.3                Dissolved Oxygen (DO)                                                                                 17

3.7       Determination of Biogas Flammability                                                                     17

3.9         Collection of Sample for Isolation                                                                           17

3.9       Media Preparation                                                                                                      18

3.10     Isolation Of Microorganisms (Fungi And Bacteria)                                                  18

3.13  Isolation And Identification Of Bacterial Isolates                                                         19

3.14  Gram Staining                                                                                                                19

3.14  Biochemical Identification of the Isolates                                                                     19

3.14.1 Catalase test                                                                                                                 20

3.14.2  Coagulase Test                                                                                                           20

3.14.3  Oxidase Test                                                                                                               20

3.14.4   Indole Test                                                                                                                 20

3.14.5 Citrate Utilization Test                                                                                                21

3.14.6 Urease Test                                                                                                                  21

3.14.7 Methyl Red Test                                                                                                          21

3.14.8 Voges- Proskauer Test                                                                                                 21

3.14.9 Sugar Fermentation Test                                                                                             21

CHAPTER 4:

RESULTS

4.0    Results of Biogas Produced and microbial isolates                                                      23

4.1    Results                                                                                                                           23

CHAPTER 5:

 DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1    Discussion                                                                                                                      29

5.2    Conclusion                                                                                                                     31

5.3    Recommendations                                                                                                         32

REFERENCES                                                                            47


 





LIST OF TABLES


Table                          Title                                          Pages

4.1       The Temperature pH, Dissolved oxygen level and conductivity

readings of the bioreactor at two days interval                         24

4.2       colonial Description and Biochemical characteristics of the bacterial isolates         27

4.3       lag period, Cumulative gas yield and mean volume of gas      28

                                                           


 





LIST OF FIGURES


Figure                                 Title                                  pages

 

1.       Conversion of fermentative substrate to biogas                                                          11

2.       Total Viable count, Anaerobic count and Total Fungal count measured in cfu/ml.    29

3.       Diagram showing fabricated bioreactor                                                                        37

 


 


 

 

CHAPTER ONE

1.1      INTRODUCTION

The demand for renewable fuel is increasing with growing concern about climate change, air quality, energy import dependence and the depletion of fossil fuels. Biogas is one of the versatile renewable fuels which can be used for power and heat/cool production or it can be upgraded to biomethane to be used as vehicle fuel. Biogas is produced by anaerobic digestion of biological wastes such as cattle dung,  cow dung, food waste, sheep and poultry droppings, plantain peels, municipal solid waste, industrial waste water and landfill 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 (Ofoefule and Ibeto, 2010). The composition of the mixture depends on the source of biological waste and management of digestion process (Yadar and Hesse, 2001).

 

The anaerobic fermentation of organic materials has long been used to generate useful resources which have been harnessed for the use of mankind (Uri, 1992; US Environmental Protection Agency, 2001). As early as the 18th Century, anaerobic process of decomposing organic matter was known, and in the middle of the 19th century, it became clear that anaerobic bacteria are involved in the decomposition process. Methanogens (methane producing bacteria) are the last component in a chain of microbes which degrade natural material and return the decay products in the environment. Hydrogen and carbon dioxide or acetate is used by methanogenic archea for the fructification of methane (also termed methanation) (Aisha and Shagufta, 2013). In this series of action biogas is generated and the methane is finally reformed to electricity. Although it is burdensome to detect attenuated organisms in very distinct communities acting as those in anaerobic digesters, defining community morphology can provide good information concerning the functional potential of the community (Aisha and Shagufta, 2013).

The energy in plant vegetation, animals, industrial and domestic waste matter can be released in terms of a useful gas when fermented anaerobically, that is in the absence of oxygen the biogas formed after the decomposition of organic wastes is channeled or transported to homes for use for cooking, running engines, electrical power generation and heating with virtually little or no pollution at all (Ozor et al; 2014). As demand for energy is increasing astronomically, and the fossil based fuels become scarce and more expensive and carbon dioxide emission levels become of more concern; Biogas a by-product of anaerobic fermentation and a renewable energy source have currently been recognized globally as a means of solving the problem of rising energy prices, waste treatment/management and creating sustainable development (Rao and Seenayya, 1994; Ofoefule and Uzodinma, 2004).

Hashimoto and Varriel, (1981), defined Biogas as a colourless, flammable gas produced through anaerobic fermentation (digestion) of animal, plant, food, human, industrial and municipal waste to produce methane (50-70%), carbon dioxide (30-40%) and traces of other gases such as hydrogen (1-10%). Nitrogen (1-3%), oxygen (0.1%), carbon monoxide and trace of hydrogen sulphide. However, the composition of the mixture depends on the source of biological waste and management of digestion process (Yadar and Hesse, 2001).

The natural generation of biogas is an important part of biochemical reaction which takes place under anaerobic condition in the presence of highly pH sensitive microbiological catalyst that are mainly bacteria (Uzodinma and Ofoefule, 2009). The production of Biogas comprises of three bio chemical processes; or steps: hydrolysis, acidogenesis/acetogenesis and methanogenesis (Nagamani and Ramasamy, (1999). Biogas is formed solely through the activity of bacteria. Bacteria have a temperature range which they are most productive in term of production rates, growth rates and substrate degradation performance. Several groups of bacteria are involved in anaerobic digestion and they also have different optimum temperature. This results in two main temperature ranges at which digestion usually can be done optimally and most economically. These ranges from 30-450C and it is termed mesophilic range and 45-700C called the thermophilic range (BTG, 2003; Ezeonu et al., 2005).

The rate of bacteriological methane production increase with digester temperature, retention time and with the percentage of total solid/volatile solid in the slurry (Dioha et al., 2006, Kalia and Singh, 1996). This is because temperature and the addition of innocula affects the enzymatic activities of the microorganisms (anaerobic) responsible for the conversion of organic materials into biogas (Kelper, 2006, Maurya et al., 1994).

One of the main environmental problems of today’s society is the continuous increase in production of organic wastes which are harmful to human existence. Some of these waste which are posing serious environmental threats to human and animal existence in these nations come from agriculture due to their degradable nature and lack of profitable technique to convert these “wastes” to better manure quality or other useful means such as energy (Okoroigwe et al., 2013).

 Cassava (Manihot exculenta) is one of the major root crops produced in sub-Saharan Africa. It is a very important crop grown for food and industrial purposes in several parts of the world. According to NPC (2008), Nigeria has the largest production of the crop of 49million tonnes of cassava. The processing of cassava results in the production of peels, chaff, fibre, and spoilt or otherwise unwanted tubers which is feed directly to ruminants. However, the much larger remaining proportion of cassava solid wastes are indiscriminately discharged into the environment and amassed as waste dumps on sites where cassava is processed. With increased production of peels and other cassava derived wastes, this constitutes an enhanced risk of pollution to the environment (Adelikan and Bangboye; 2009). This is also a need to find an alternative productive use of the peels. One area of possibility is to investigate the potential of cassava peels for the production of biogas and by so doing, it will reduce the nuisance value it creates in the environment.


1.2       AIM AND OBJECTIVES

This study was aimed at:

1.     Producing combustible biogas from cassava peels

2.     Isolating and characterizing the microbes responsible for the biodegradation of cassava peels in biogas production.

3.     Evaluating the physicochemical parameters of the slurry during biogas production

 

 

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