ABSTRACT
The distribution and characterization of a suspected petroleum bearing site (SPBS) is generally called microbial prospecting. Microbial surface prospecting is a vital process in the exploration of hydrocarbon suspected site. This study was aimed at assessing the distribution, characterization and hydrocarbon degrading potentials of microorganisms in some parts of Bida Basin, Nigeria. The study assessed the physicochemical and microbiological properties of soil and water from Ahoko, Kogi State, and Patishabakolo, Niger state, Nigeria, to ascertain the potent site for exploration using the presence of microbial anomalies. Standard plate count, ArcGIS, gravimetric and GC- MS methods were used to ascertain the total bacterial and total fungal counts, distribution of specific hydrocarbon–oxidizing microbes and hydrocarbon-degrading potential of isolates respectively. Physicochemical properties showed that pH of the soil samples were within the range of 5.58 – 7.05. Site A had the highest total organic carbon (TOC) (1.18%), conductivity (12.0 μ∫/cm), available Phosphorus (12.06 mg/kg), base saturation (66.03 ± 0.00%) and exchangeable acid (2.28 cmol/kg) compared to other sites in Ahoko. The physicochemical parameters such as pH, TOC, conductivity and total pore space in samples from Patishabakolo vary from 5.30-6.81, 1.75%-8.78%, 0.10µʃ-2.0µʃ/cm and 160.8%-192.3% respectively. The study revealed the presence of counts for methane-oxidizing microbes (MOM) (4.8 x 102 CFU/mL, 5.0 x 102 CFU/mL), ethane-oxidizing microbes (EOM) (4.6 x 102 CFU/mL, 10.0 x 102 CFU/mL) propane-oxidizing microbes (POM) (3.6 x 102 CFU/mL, 10.0 x 102 CFU/mL) and Butane oxidizing microbes (BOM) (2.80 x 102 CFU/mL, 6.65 x 102 CFU/mL) in water samples from Ahoko, and Patishabakolo respectively. There were significant differences (p<0.05) in counts of the various groups of bacteria except for BOM. In general, the EOM predominated in all study sites and areas. The EOM was the most widely distributed across all the study locations and there were correlative relationship (r = 0.9) between EOM and BOM. The most frequently encountered bacterial and fungal hydrocarbon-gas utilizers were Corynebacteria, Acinetobacter, Micrococcus and A. niger and A. flavus respectively. Ten crude oil–utilizing organisms which included one bacterium and 9 fungi were amplified using using universal primers. The organisms were identified as, Enterobacter asburiae RSo-A, Aspergillus flavus A-Feb, A. tubingensis D2S3, Alternaria tenuissima JYW, A. flavipes BH A. terreus JYS, Penicillium pimiteouiense JS-B, Aspergillus sublatus D1S4, Aspergillus stellatus Borehole, and Candida orthopsilosis JS-A. The rate of crude oil degradation by the identified organisms was determined after 35 days of incubation, using gravimetric analysis method. The results revealed that 68.9 %, 73 %, 68.4 %, 62 %, 72.11 %, 77.8 %, 83.68 %, 68.89 %, 88.05 %, and 85 % degradation were respectively achieved by the organisms. Biodegradation potential of isolates were observed by conversion of high molecular weight (HMW) compounds to low molecular weight (LMW) compound, reduction in the height of peaks and the concentrations of carbon content as well as presence and emergence of intermediates compounds. All the isolates used in this study utilized the crude oil luxuriantly but isolates of C. orthopsilosis JS-A and A. stellatus borehole were the most effective and efficient degraders, achieved 85 % and 88 % rate of degradation and was able to completely remove 9 and 13 carbon compounds from the crude oil mixture respectively. Hence, they can be used as clean up agents in oil spills in future if oil exploration begins in these zones.
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background to the Study
One of the suspected petroleum bearing sites in Nigeria is in the Bida Basin, which is located in the North Central Region, Nigeria. Ladipo (1998) reported that the Basins sedimentary fill consists of post-orogenic molasses and slender unfurled marine residue. It is characterised by means of the absence of carbonates, volcanic, and rocks of tertiary ages (Rahaman et al., 2019). The basin has an inverted arch due to fracturing that originates close to Santonian orogenic tendencies in South East Nigeria and the Benue valley (Obaje et al., 2011; Usman, 2019).
Petroleum forms through the setting apart of large debris of waxes, oils and fats that associates with the formation of kerogen (Mansoori et al., 2016). This system commenced extraordinary a few years back, whilst little marine lifestyles bureaucracy possessed big quantities of the oceans died, they settled at the sea depths and ended up blanketed in layers of dirt, sediment and sand. The non-stop rot via way of means of the effect of heat and strain pressure delivered approximately the improvement of many additives of oil compounds (Mansoori et al., 2016). Petroleum is a liquid, due to that, it could move via the earth as its forms.
Oil has mostly been recouped via means of oil dull that is achieved after investigations of auxiliary topography (on the repository scale), sedimentary bowl examination, and shop portrayal were finished (Guerriero et al., 2012). One of the strategies utilized for hydrocarbon investigation, portrayal and distinguishing proof of microorganisms from hydrocarbon presumed site is regularly known as microbial prospecting and it depends on the way vaporous hydrocarbons through emission and diffusion migrate upward from subsurface petroleum accumulations, and are utilized by a variety of microorganisms present in the sub-soil ecosystem (Etiope, 2015; Hubert and Judd, 2020). The methane, ethane, propane, and butane-oxidizing bacteria exclusively use these gases as carbon source for their metabolic activities and growth. These microorganisms are mostly found enriched in the shallow soils and can differentiate between hydrocarbon prospective and non-prospective areas (Rasheed et al., 2014).
Biodegradation is characterized as the naturally catalyzed decrease in multifaceted nature of substance mixes (Joutey et al., 2013). It is the procedures that make use of microbes to breakdown large compounds into smaller mixes (Marinescu et al., 2009). In biodegradation, living things change the substance through metabolic or enzymatic procedures which depends on development and co-digestion (Fritsche and Hofrichter, 2008). A few microorganisms, including parasites, microscopic organisms and yeasts are engaged with biodegradation process. The reports for algae and protozoa are meagre with respect to their association in biodegradation (Das and Chandran, 2011; Joutey et al., 2013). Biodegradation by natural populations of microorganisms represents one of the primary mechanisms by which petroleum and other hydrocarbon pollutants can be removed from the environment and is much less expensive than different remediation innovations (Ulrici et al., 2000; Abioye et al., 2013). Biodegradation of hydrocarbons begins by the conversion of the alkane chain or polycyclic aromatic hydrocarbon (PAH) into an alcohol. The next step in the process is oxidation, which then converts the compound to an aldehyde and thereafter, into an acid and eventually into water, carbon dioxide, and biomass. In the case of the PAH, fission occurs which leads to mineralisation (Shallu et al., 2014). The process of bioremediation, defined as the use of microorganisms to detoxify or remove pollutants owing to their diverse metabolic capabilities is an evolving method for the removal and degradation of many environmental pollutants including the products of petroleum industry, this technology is believed to be non-invasive and relatively cost-effective (Rasheed et al., 2015).
The success of oil spill bioremediation depends on one’s ability to establish and maintain conditions that favour enhanced oil biodegradation rates in the contaminated environment. Numerous factors influence the rate of oil biodegradation. One important requirement is the presence of microorganisms with the appropriate metabolic capabilities. If these microorganisms are present, then optimal rates of growth and hydrocarbon biodegradation can be sustained by ensuring that adequate concentrations of nutrients and oxygen are present and that the pH is between 6 and 9. The physical and chemical characteristics of the oil and oil surface area are also important determinants of bioremediation success (Das and Chandran, 2011).
1.2 Statement of the Research Problem
The Bida and Sokoto Basins are the only Basins in the country yet to produce oil blocks for auctioning by the Federal Government. With the increase in demand for petroleum products and diminishing indigenous production, it has become necessary to search for probable potential zones or areas with the aim of expanding the national exploration and production base and adding to the proven reserves. In addition to the search for other potential area for exploration, the problem of hydrocarbon contamination known to be widespread in oil producing areas should not be overlooked.
One of the major environmental problems today is hydrocarbon contamination resulting from the activities related to the petrochemical industry with accidental release of petroleum products being a particular concern in the environment. Hydrocarbon components have been known to belong to the family of carcinogens and neurotoxic organic pollutants (Das and Chandran, 2011). Currently, accepted disposal methods of incineration or burial in secure landfills can become prohibitively expensive when amounts of contaminants are large. Besides, mechanical and chemical methods generally used to remove hydrocarbons from contaminated sites have limited effectiveness and can be expensive (Malik and Ahmed, 2012; Abioye et al., 2013). Bioremediation is the promising technology for the treatment of these contaminated sites since it is cost-effective and will lead to complete mineralization. Bioremediation functions basically on biodegradation, which may refer to complete mineralization of organic contaminants into carbon dioxide, water, inorganic compounds, and cell protein or transformation of complex organic contaminants to other simpler organic compounds by biological agents like microorganisms (Malik and Ahmed, 2012).
1.3 Justification for the Study
The Bida Basin is one of the inland Basins in Nigeria that is located in the North Central and extends to the West-Central region of Nigeria. Through geological survey, Bida Basin has been suspected to have the potential for an active petroleum system with the generation, migration and accumulation of hydrocarbons in commercial quantities (Obaje et al., 2011). There is therefore the need for microbiological studies to prove that this Basin indeed can be explored for petroleum. Hence this study was directed to identify the presence and distribution of hydrocarbon degrading microorganisms in the basin. Besides, in oil exploration site there is always pollution, if exploration begins in this basin pollution will be inevitable, for effective remediation of oil polluted site, suitable microbial inoculants as clean up agents in oil spill incidences should be formulated. Suitable microbial inoculants or soil microcosm cannot be efficiently formulated if the microorganisms are not characterized and identified (Tharaka et al., 2016), therefore characterization of isolates is a very important aspect in order to develop a successful bioremediation strategy for petroleum oil contaminants. In addition, identifying the degradation capacity of these organisms helps in constructing more competent oil degraders for oil spill remediation.
1.4 Aim and Objectives
The aim of the study was to assess the distribution, characterization and hydrocarbon degrading potentials of microorganisms in some parts of Bida Basin, Nigeria.
The objectives of the study were to:
i. determine the physicochemical and microbiological properties of soil and water sources in the suspected petroleum bearing sites (SPBS)
ii. characterize and identify the microbial isolates
iii. study the distribution of the hydrocarbon utilizing microorganisms in the SPBS.
iv. determine the petroleum biodegradation potential of the microbial isolates
This material content is developed to serve as a GUIDE for students to conduct academic research
DISTRIBUTION, CHARACTERIZATION AND HYDROCARBON– DEGRADING POTENTIALS OF MICROORGANISMS IN SOME PARTS OF BIDA BASIN, NIGERIA
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