ABSTRACT
Biodegradation has proved over time that it is the cheapest and safest method human can use to tackle waste and population. The study of biodegradation of polypropylene revels that Actinomycyte and three other unknown strains are capable of biodegrading polypropylene (making new functional group) within 3weeks. The new functional group seen after 3 weeks were ester, cyanide, and ketone. The microbial community at the Yola waste are a community of diverse organism. Each with its own unique morphology, and growth pattern. All organisms were gram positive. This means that they can adapt to high stress and a resist turgor pressure. Actinomycyte spp was able to biodegrade polypropylene by breaking the carbon to hydrogen, and breaking down carbon hydrogen bond to make carbon oxygen bonds.
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TABLE OF CONTENTS
CERTIFICATION…………………………………………………………………….ii
READERS’APPROVAL………………………………………………………………iii
DEDICATION………………………………………………………………………….iv
ACKNOWLEDGEMENTS……………………………………………………………..v
ABSTRACT……………………………………………………………………………vi
LIST OF TABLES……………………………………………………………………ix
LIST OF FIGURES……………………………………………………………………x
CHAPTER 1………………………………………………………………………….1
INTRODUCTION……………………………………………………………………..1
Waste……………………………………………………………………………………1
Composition……………………………………………………………………………….2
Waste management ………….…………………………………………………………4
Bioremediation…………………………………..……………………………………5
Degradation of waste materials by microorganisms………………………………….7
Keratin degradation…………………………………………………….……………7
Plastic degradation………………………………………………………………………………8
Anaerobic digester……………………………………………………………..……..9
Types of bioremediation…………………………………………….……………….11
Compositing …………………………………………………………………………11
Bioventing ……………………………………………………………………………………12
Case of Nigeria………………… …………………………………………………………….13
AIMS & OBJECTIVES…………..……………………………….………………..15
CHAPTER 2……………………………………………………………………….16
MATERIALS & METHODS………………………………………………16
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Study site………………………………………………………………………16
Sampling techniques……………………………………………………….17
CHAPTER 3………………………………………………………………………..20
RESULTS…………………………………………………………………..20
CHAPTER 4………………………………………………………………………….25
DISCUSSION…………………………………………………………….…25
CHAPTER 5…………………………………………………………………………33
CONCLUSIONS AND RECOMMENDATIONS……………………………………………………………33.
REFERENCES…………………………………………………………………….. 34
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LIST OF TABLES
Table 1: IR interpretation chat for all sample after 2 week of biodegradation……………………………………………………………………….22
Table (2). Waste dump soil organism biochemical test……………………………………………………………………………………23
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LIST OF FIGURES
Fig:1 Composition of household wastes………………………………………………2
Fig:2 Municipal waste collection data for different countries………………………..4
Fig:3 Different bioremediation techniques used across the world………………………………………………………………………………13
Fig:4 Map of Yola town………………………………………………………………..16
Fig:5 IR spectrometer reading of Polypropylene (PP): B untreated (un-inoculated), A, C and D treated samples (inoculated with strain 1 and 2 for 2weeks in a broth of PP and water ………………………………………………………………….…….…..21
Fig:6 Colour change due to pseudomonas spp………………………….…………..23
Fig:7 Microscopic view of bacteria found in the total sample soil at total magnification of 1000X……………………………………………………………………………..24
Fig:8 Molecular structure of polypropylene…………………………………………27
Fig:9 Molecular structure of Ester……………………………………………………28
Fig.10 Molecular structures of aldehyde and ketone…………………………………………..28
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CHAPTER 1
INTRODUCTION
Every year, about 1.3 billion tons of waste is generated globally. By 2025, it is estimated that this will increase to 2.2 billion tons, with a large sum (2.13 kg/capita/day) from developing countries (Bhada-Tata, &Hoornweg, 2012). The term waste refers to items or products (organic and inorganic) that are considered useless or have lost their value (Bhada-Tata, & Hoornweg, 2012). It is a broad term items such as animal bones, plastic bags and bottles, and used clothes. Waste generation has been in existence, since the beginning of agricultural revolution, and it can be traced back to the first human civilization. As a result, waste is inevitable and cannot simply be avoided, due to urbanization (Muhammad, Huma, Munir, & Atiq, 2015). Information age, urbanization and industral age has led improve human lifestyle, in many cities, which is the major cause of increase in solid waste production (Renou, 2008).
With the emergence of the industrial age, and then the information age, urbanization could not stifled because of the human desire for securing a more comfortable life. This led to the production of waste products. Urbanization generally brings economic prosperity, and higher waste production. This is because people living in cities usually earn more income, and have several options from which they can choose from (Renou, Givaudan, Poulain, Dirassouyan, & Moulin , 2008).However, urban settlements are known to be densely populated, leading to greater amount of waste compared to non-urban areas(Renou, Givaudan, Poulain, Dirassouyan, & Moulin , 2008).
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Composition of waste in urban areas
On average, urban waste dumps consist of millions of different waste materials (Bhada-Tata, &Hoornweg, 2012). The composition of a landfill or dumpsite gives an idea of the physical, chemical, and thermal properties of the waste. There are four factors that influence the composition of waste in a dumpsite: culture, seasons, laws guiding waste disposal, and demographics of people living in the area. For example, in developed countries and in urban areas people tend to consume more processed foods than unprocessed food. (Bhada-Tata, & Hoornweg, 2012).
Municipal wastes are garbage collected and transported to landfills from households and industries. Municipal waste in landfills serve as a home and substrate for microorganisms and provide a unique ecosystem for compositing and anaerobic digesters.
Fig.(1). Composition of household wastes in the world (Ogola, Chimuka, & Tshivhase, 2011)
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The major challenge in the today’s world is that most of the waste generated are inorganic, and the rate of degradation is inversely proportional to the amount of it produced (Pettigrew, Palmisano, & Charles , 1992). In other words, the rate at which our consumer-driven society utilizes and discards products, especially inorganic ones, supersedes the time needed for the products to decompose. This situation is obvious and can be found in the open garbage dumpsites in developed, developing, and under-developed countries. In developed countries such as United States of America, 25 million tons of plastics are discarded every year. Most of the plastics are deposited in landfills where the degradation process may last for decades or centuries, therefore leading to difficulties in locating new landfill sites (Pettigrew, Palmisano, & Charles , 1992).
Open landfills are rapidly increasing due to the rate at which the world is producing and discarding products, which has led to the extension of existing garbage sites into new lands. New garbage sites are also being created, and this is a threat to agriculture because it takes up land that could have been used for the cultivation of crops. An issue with garbage dump sites is that it causes environmental pollution, such as soil contamination and air pollution, which eventually leads to water contamination and human health hazards such as diarrhea, respiratory ailments and dengue fever (Agnieszka Kalwasińska, 2012).
In ancient times, when mankind relied on foraging for survival, waste management was not a major issue because the small human population produced little waste compared to that which we produce in tons daily (Giusti, 2009). The wastes we produce are poorly managed globally, especially in developing countries, which is
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causing serious environmental pollution (Giusti, 2009). The impact of improper waste disposal can be seen in developing countries around the world.
Waste management in developing countries
Composition, waste generation, and the waste management practices in use vary from one geographical area to another in both developed and developing countries (Vaibhav, & Sultan, 2014). Due to the pollution caused by poor waste management practices, there has been concern about control practices, inadequate legislation, and the environmental and human health impacts of waste.
As a result, some countries have made efforts to enact laws that control unsustainable
Fig. (2).
Fig. (2). Waste collection data for different countriesWaste collection data for different countries
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waste management. However, in developing countries globally, waste may be dumped in open landfills, without adhering to the recommended rules by the municipal authorities. Landfills are lands where solid waste is disposed. They are usually located at the outskirts of urban centers. Landfills are usually the final site for waste disposal in many developing countries. This is because open-dump sites are cheap and affordable for any nation (Vaibhav, & Sultan, 2014). However, in developing countries, few data exist about open waste dumps, and this could be a reason for continuous environmental pollution. . In developed countries on the other hand, efforts have been made in reducing the amount of municipal wastes littered in the environment. Developed countries have made modern waste management models. These models include modern recycling, incineration, and anaerobic digestion has been developed. In addition, countries have begun investigating a natural form of waste clean-up: bioremediation.
Bioremediation
Bioremediation is a naturally occurring process whereby micro-organisms convert harmful products to less toxic products (Arvanitoyannis,&Thassitou, 2001).Municipal waste acts as substrate for many microorganisms. These micro-organisms provide a unique ecosystem for composting and anaerobic digesters. Each layer of a landfills provide a conducive environment for microorganisms (Palmisano & Barlaz, 1996).
The upper layer has the most nutrients because of the adequate moisture and temperature present. A typical waste dump consists of polymeric substances, such as paper, yard waste, and food. Therefore, waste dumps act as substrates for microbe ,
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and these substrate will include cellulose, starch, protein, and hemicellulose. The microbial organisms in landfills include digesters, composters, anaerobic digesters (hydrolytic and fermentative bacteria), acetogen, methanogens and sulphate reducers. Other micro-organisms present include such bacterial species as Bacillus spp., Escherichia coli, Klebsiellaspp., Proteus spp., Pseudomonasspp., Staphylococcusspp., and Streptococcus spp. On the other hand, fungi such as Aspergillus, Fusarium, Mucor, Penicillium, and Saccharomyces are capable of biodegrading waste materials (plastics). All these microbes work together to breakdown waste as part of the bioremediation process.
Bioremediation has been found to be the most sustainable way for water and soil remediation. This is because it does not pose a threat to the environment or human health, and it is inexpensive (Arvanitoyannis, & Thassitou, 2001). Bioremediation processes for the treatment of contaminated soil and water are divided into four categories: inoculation, stimulation, use of immobilized enzymes, and use of plants. The methods used are composting, landfarming, use of bioreactors, and intrinsic bioremediation (Arvanitoyannis,& Thassitou, 2001).There are different types of bioremediation that help restore polluted sites. The total number of organisms found in a polluted area, gives an insight on how efficient the wastes can be degraded over time. In addition, the method implore for the biodegradation also count to how fast the waste is degraded over time. These methods includes gravimetric method and the ohimic technology. The gravimetric method is an old method of degrading waste. However, advanced technologies, such as the ohmic technology, have helped in overcoming the limitations associated with characterisation of wastes for biodegradation (Azubuike, Chikere, & Okpokwasili, 2016).
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Bioremediation of waste materials Biodegradation of Agricultural wastes Keratin biodegradation Keratin, fibrous structural protein of hair, nails, horn, hoofs, wool, feathers, and of the epithelial cells in the outermost layers of the skin. Keratin serves important structural and protective functions, particularly in the epithelium. It is durable because of the cross-linkage bond between disulphide and hydrogen bonds. It is an insoluble protein. Keratins are made up of amino acids, including cysteine, lysine, proline, and serine(Călin, et al., 2017). However, keratin is classified into beta (hair and wool) and alpha sheet (sheet of feathers). It can be also grouped into hard or soft keratin (Jeffrey et al, 1995).
Keratin waste products are a threat to the environment. However, they are release in large quantity by the agricultural industries, in the form of nails, horns, and feathers of animals. Keratin has a high degree of stability, and can be difficult to degrade. However, only organisms that can secrete keratinocytic enzymes, such as keratinases, can degrade keratin (Mariana et al., 2017). Many microorganisms grow on keratinous materials; Bacillus spp. can grow on and biodegrade keratin. The biodegraded product can be use as feeds for chicken, fertilizers, etc (Jeffrey et al, 1995).
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Plastic degradation
Plastics are of two types: thermoplastic and thermoset. Thermoplastics are formed by the breaking the double bonds found in olefin (polymerization), leading to a formation of carbon-carbon bonding. Thermosets are produced by the removal of water from a bond between carboxylic acid and an amine or alcohol (Ying Zheng,& Ernest K. Yanful, 2005). Plastics are polymeric molecules characterized by long chains (Ying, Yanful, & Bassi, 2005).
Plastics are materials that can be moulded into different shapes. Although plastics are characterized as soft, they can harden when exposed to certain factors, such as temperature (Pettigrew, Palmisano, & Charles , 1992). After been transformed to different shapes, plastics become stable and are difficult to degrade. Some plastics such as Polyolefins are considered non-biodegradable because of their hydrophobic characteristics, high molecular weight, and use of anti-oxidants (Ying, Yanful, & Bassi, 2005).
Thermoplastic degradation
Polyolefins are thermoplastics that consist of one-carbon backbone, and they are classified as non-biodegradable. This is because of their large molecular mass. However, studies have shown that microorganisms can degrade thermoplastics with low molecular mass. This is done by integration of starch such as glycerol plasticized starch with microorganism (Keiko, Hiroshi, Yuhji , & Tani, 2001). For a plastic to biodegradable, it must have a molecular mass of less than 500g/mole(Keiko, Hiroshi, Yuhji , & Tani, 2001). However, the molecular mass of most plastics ranges from 40,000 to 28,000g/mole Keiko, Hiroshi, Yuhji , & Tani, 2001). For microorganisms to biodegrade plastics, their molecular mass must be lowered. This is done modifying
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their structure by adding a special form of synthetic Polyolefins. This structure can then be broken down by photo-degradation, and chemical degradation leading to short chains of carbon. Therefore, reducing the molecular mass to the acceptable value ofless than 500 g/mol will enable biodegradation. With the molar mass of less than 500 g/mol, microorganisms can further biodegrade polyolefins monomeric and oligomeric: photo-degradation and chemical degradation of plastics(Keiko, Hiroshi, Yuhji , & Tani, 2001).
Thermoset degradation
Thermoset plastics are of two types: polyurethane and polyesters. According to Ashak,& Rathoure direct enzymatic attack on polyesters had no effect. Strains of Trichosporum and Arthrobacter has been effective in degrading polyester (Ying Zheng & Ernest K. Yanful, 2005). Bioremediation of polyesters by Trichosporum and Arthrobacter occurred within one week. Polyurethanes are used in furniture and paint. Microorganisms degrade polyurethane by attacking the urethane component. There are three types of polyurethane bio-degradation: fungal biodegradation, bacterial biodegradation, and degradation by polyurethanase enzymes. Examples of fungi that perform bioremediation of polyurethane are Curvularia senegalensis, Fusarium solani, Aureobasidium pullulans,and Cadosporium sp.. However, 100% bioremediation of polyurethane has not been achieved (Ashak,& Rathoure, 2016).
Anaerobic digestion
Anaerobic digestion is a process whereby microorganisms break the waste in the absence of oxygen. This process requires amicrobial community of facultative organisms. These facultative organisms decompose waste such as food (protein,
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carbohydrate, and lipids). The process begins with the hydrolysis of solid waste. Solid waste is decomposed into smaller masses (monomers or dimers) by hydrolytic organisms. These organisms can degrade soluble and non-soluble materials because they can produce hydrolytic enzymes. The monomers or dimers are further fermented into hydrogen, carbon, or fatty acids by acetogens and methanogens(Maritza et al., 2008).
Other organisms involved in bioremediation of food waste are nitrate-reducing bacteria, methanosarcina, and methanothrix. Materials that can be biodegraded by anaerobic digester are waste food (rice, beans, yam, and so on) and paper. The anaerobic digester method is effective. However, it becomes a problem if the microbial organisms are unequally distributed (Alvarez, 2003). This method was carried out in Mexico for the purpose of turning waste into organic manure in the absence of oxygen. The major components of the digested waste were 62% paper, 23% food waste, and 15% yard clippings (Călin, et al., 2008). In this study, it was observed that the rate of degradation is dependent on the number of organisms present.
Bioremediation in Developing Nations
Bioremediation of contaminated soil, which includes open landfills (consisting of keratin, plastics and food wastes), soil contaminated with oil, and water contamination is increasingly gaining audience around the world. However, this is mostly seen in developed countries such as United States of America. Developing countries are yet to embrace this technology, because of the lack of technologies, and environmental regulations that favours bioremediation. The highest rate of bioremediation comes
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from developed nations. Although, efforts have been made by developing countries to embrace the technology, however, however, their efforts are minimal. For example, America (Canada and United States of America) a developed nation has implore 61.85%, of bioremediation while developing countries such as China (7.9%) Japan (6.77%), Korea (4.5%) and India (2.93%). India as a developing nation has shown a great zeal towards the use of bioremediation techniques ( Saraswat , 2014).
Research on bioremediation and the use of bioremediation techniques are not only seen in Asia (developing nation), but are also seen in some African countries.
Bioremediation of contaminated soil and water have been carried out in countries such as South Africa. In South Africa bioremediation of water contaminated with metals was carried out in Cape Town, and isolation and characterization of engine oil degrading indigenous microorganisms in Kwazulu-Natal (Ogola, Chimuka, & Tshivhase, 2011). In Uganda, which is in eastern Africa bioremediation techniques are used in cleaning up water contaminated with petroleum. In Nigeria, bioremediation has been used in cleaning up water and land contaminated with petroleum. However, little or no research has been conducted using bioremediation to clean up open landfills in Nigeria as well as the whole of Africa (Ogola, & Tshivhase, 2011).
Methods for Bioremediating waste in Developing Nations
Composting
Compositing is a bioremediation method used in remediating waste dumps in countries such as India. Composting is the use of microorganisms to decompose organic materials using aerobic respiration. In this process, organisms and organic
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materials are kept under high temperature, which enables microbes to degrade waste efficiently. This process occurs in nature, but at a slow rate, and material decomposition is hardly noticed (Arvanitoyannis,& Thassitou2001). This decomposition process is seen in waste dumps. To efficiently clean up waste, microbial growth must be induced. The process starts with introducing an organism, which is most efficient at moderate temperature of 300C to 450C (Arvanitoyannis, &Thassitou, 2001). The organisms increase the temperature by 50C to 60C setting a temperate for the growth of thermophile organisms, which are heat loving organisms (Arvanitoyannis, & Thassitou, 2001). Thermophile organisms work better at higher temperature and are continually increase the temperature around their environment. Therefore, for maximum growth and efficient degradation to be achieved, the temperature should be carefully monitored to avoid it browning up to 700C (Arvanitoyannis,& Thassitou, 2001). Examples of composting microbes are acetogens and methanogens. These organisms are used in biodegradation of plastics (Arvanitoyannis, & Thassitou, 2001).
Bioventing
In Africa, the bioremediation process use in cleaning up soil pollutant is bioventing. Bioventing is a bioremediation process of introducing oxygen to unsaturated polluted site, leading to the growth of microbes. Nutrients and moistures are introduced into the polluted soil, and this allows the growth of bioremediation organisms. The process is more efficient when the water tables are dip within the soil structure (Andrea, & Hinchee, 1997)
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Fig (3): Different bioremediation techniques used across the world( Saraswat , 2014)
Case of Nigeria
It is estimated that by 2050, Nigeria will become the 3rd most populous country, and land will be needed for agriculture and other purposes ( United Nations, 2017). Due to poor waste management in Nigeria, open waste dumps are greatly increasing. However, most of the waste takes years or centuries to degrade naturally, and the finally degraded pollutants are emitted, which leads to health hazards.
However, bioremediation may be a valuable tool for developing countries such as Nigeria for managing waste. This will lead to a healthier environment and better human health. The efficiency of bioremediation may vary from site to site and is dependent on the types of microorganisms present and microclimate factors, such as humidity, soil type, and temperature. Therefore, I investigated the potential of microorganisms found in open dumpsites in Jimeta-Yola in north-eastern of Nigeria to perform bioremediation was investigated. The study was aimed to determine what
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microorganisms are present in the soil in the selected site and how well they perform bioremediation on solid waste.
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AIMS & OJBECTIVES Aims To investigate the ability of microorganisms to perform bioremediation on solid waste from dump sites in Jimeta-Yola, north-eastern Nigeria
Objectives
• To collect soil samples from different segments of the waste dump in Jimeta -Yola
• To culture the isolated microorganisms.
• To identify the types of microorganisms found in the solid waste dump site.
• To isolate microorganisms with solid waste materials.
• To evaluate degradation of waste over time by each microorganism
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