scholarly journals Characterization of Indigenous Bacterial Communities in Crude-Oil Impacted Sites at Obagi Town, Onelga, Rivers State, Nigeria

Fine Focus ◽  
2016 ◽  
Vol 2 (1) ◽  
pp. 7-14
Author(s):  
Chioma Blaise Chikere ◽  
Chinyere Augusta Ajuzieogu ◽  
Michael Chukwugoziem Miller

Hydrocarbon utilizers are expected to be indigenous in crude-oil polluted environments. The isolation and characterization of hydrocarbon utilizers is often a key strategy in bioremediation of hydrocarbon-polluted environments. In this study, crude-oil polluted soil samples from Obagi town, Onelga, Rivers state were enumerated and characterized for putative hydrocarbon utilizing bacterial populations. Biochemical characterization identified five bacterial species representative of five genera: Bacillus, Pseudomonas, Acinetobacter, Micrococcus and Staphylococcus. Amongst the genera of bacteria isolated, Bacillus had the highest frequency of occurrence (40%). The mean count of total heterotrophic bacteria was 1.7 X 107 cfu/g, while hydrocarbon utilizing bacteria (HUB) count mean density was 1.0 X 107 cfu/g for the three soil samples. Statistical analyses revealed no significant difference at p>0.05 between Total Heterotrophic Bacterial (THB) and Hydrocarbon Utilizing Bacterial (HUB) counts, suggesting that most of the bacteria present in the sampled sites were hydrocarbon utilizers. Findings from this study suggest the presence of indigenous putative hydrocarbon utilizing bacteria in the crude-oil polluted soil of Obagi town. Hence, a promising potential exists for future bioremediation studies on the site.

Author(s):  
D. N. Ogbonna ◽  
S. A. Ngah ◽  
R. N. Okparanma ◽  
O. Ule ◽  
R. R. Nrior

Aim: The aim of the study was to assess Percentage Bioremediation of Spent Mushroom Substrate (SMS) and Mucor racemosus in hydrocarbon contaminated soil Place and Duration of Study: A portion of Rivers State University demonstration farmland in Nkpolu-Oroworukwo, Mile 3 Diobu area of Port Harcourt, Rivers State was used for this study. The piece of land is situated at Longitude 4°48’18.50’’N and Latitude 6o58’39.12’’E measuring 5.4864 m x 5.1816 m with a total area of 28.4283 m2. Bioremediation monitoring lasted for 56 days, analysis carried out weekly (per 7 days’ interval). Methodology: Five (5) experimental plots employing the Randomized Block Design were used each having dimensions of 100 x 50 x 30 cm (Length x Breadth x Height) = 150,000cm3. Baseline study of the uncontaminated and the deliberately contaminated agricultural soil was investigated for its microbiota and physico-chemical properties. Two of these plots were designated as pristine (Unpolluted soil) (CTRL 1) and crude oil contaminated soil without nutrient organics and bioaugmenting microbes (CTRL 2); these two serve as controls. Each of the experimental plots, except the control (CTRL 1), was contaminated with 2500 cm3 (2122.25 g) of crude oil giving initial Total Petroleum Hydrocarbon (TPH) value of 8729.00 mg/kg. The crude oil polluted soil in Plot 3 was further treated with 750 ml of Mucor racemosus broth (CS+Muc), Plot 4 was treated with 3000 g of Spent Mushroom Substrate (CS+SMS) while plot 5 was treated with the combination of both (CS+Muc+SMS). The plots were left for 7 days to ensure even distribution and soil-oil bonding. Sampling was done at seven-day interval (Day 1, 7, 14, 21, 28, 35, 42, 49, 56).  Physicochemical parameters monitored were pH, Temperature, Nitrogen, Phosphorus, Potassium, and Total Petroleum Hydrocarbon (TPH) throughout the experimental period. Microbial parameters monitored were Total Heterotrophic Bacteria (THB), Total Heterotrophic Fungi (THF), Hydrocarbon Utilizing Bacteria (HUB) and Hydrocarbon Utilizing Fungi (HUF). Percentage (%) Bioremediation was estimated from percentage (%) reduction of Total Petroleum Hydrocarbon (TPH) from day 1 to day 56 in relation to control plots.  Net % Bioremediation were also assessed to ascertain the actual potential of treatment agents singly or combined. Results: Total Heterotrophic Bacteria (THB) (CFU/g) recorded on day 7 and day 56 of the bioremediation were; day 7; CTRL 1 – US (1.07 x109), CTRL- CS (5.4 x108), CS+Muc (3.0 x108), CS+SMS (4.6 x108) and CS+Muc+SMS (5.0 x108). On day 56, data obtained were CTRL 1 –US (9.4 x108), CTRL 2 –CS (7.2 x109), CS+Muc (3.7 x108), CS+SMS (8.1x108) and CS+Muc+SMS (6.8 x108). The increase in number in the treated plots is a depiction of an increase in activity of the organism and the stimulating effect of bio-organics SMS while the untreated plot CTRL 1-US showed decrease in population at day 56. Similar trend showed for Total Heterotrophic Fungi. Generally, it was observed that the highest growth/ count was recorded at the 7th and 8th week (day 42 or day 49), at the 9th week there was an observable decrease; probably due to depletion of nutrients and other factors such as rainfall and seepage. The Net Percentage Hydrocarbon Utilizing Bacteria and Fungi (Net %HUB and Net %HUF) were highest in Crude Oil contaminated plot treated with Spent Mushroom Substrate (SMS) singly; that is (CS+SMS) (11.02% and 12.07%) and lowest in the uncontaminated soil – Control (CTRL 1 –US) (5.41% and 9.26%) respectively. The trend in decreasing order of Net % Hydrocarbon Utilizing Bacteria were as follows: CS+SMS (11.02%) > CS+Muc+SMS (10.14%) > CS+Muc (9.43%) > CTRL 2 –CS (8.1%) > CTRL 1 –US (5.41%) while Net % Hydrocarbon Utilizing Fungi followed similar trend and were: CS+SMS (12.07%) > CS+Muc+SMS (11.76%) = CS+Muc (11.76%) > CTRL 2 –CS (11.05%) > CTRL 1 –US (9.26%). Evaluation of Amount of Crude Oil or Hydrocarbon remediated and Net %Bioremediation revealed Crude Oil contaminated plot augmented with Mucor racemosus broth singly (CS+Muc) as having the highest bioremediation potential while the least is the untreated soil. The trend is as follows:  CS+Muc (8599.19 mg/kg; 33.93%) > CS+Muc+SMS (8298.95 mg/kg; 32.74%) > CS+SMS (8197.03 mg/kg; 32.34%) > CTRL 2 –CS (166.54 mg/kg; 0.66%) > CTRL 1 –US (85.48 mg/kg; 0.34%) Conclusion: This shows that a single nutrient substrate or augmenting microorganism applied appropriately may have a more positive result, that is; higher bioremediation potential than combined or multiple mixed treatments. It was further observed that microbial counts decreased with time in treatments with augmenting organisms alone but increased considerably in treatments supplement with organics having its peak on the 49th day.   It is therefore recommended that bioremediation of crude oil-polluted soil using bio-augmenting microorganism should be applied appropriately noting the volume: area ratio and be supplemented with efficient nutrient organics after every 49-day interval.


2020 ◽  
Vol 2 (2) ◽  
pp. 12-16
Author(s):  
Obhioze Augustine Akpoka

The capability of indigenous bacteria and microalgae in crude oil effluents to grow in and utilize crude oil as their sole source of carbon and energy provides an environmentally friendly and economical process for dealing with crude oil pollution and its inherent hazards. In view of the toxicity of crude oil spillages to indwellers of the affected ecosystems and the entire affected environment, the isolation of pure bacterial and microalgae cultures from crude effluents is a step in the right direction, particularly for bio-augmentation or bioremediation purposes. The total heterotrophic bacteria count and hydrocarbon utilizing bacteria count, as well as the microalgae count, were determined with the pour plate technique. The physicochemical properties of the effluent samples were also analyzed. Identification of the hydrocarbon utilizing bacteria was performed with phenotypic techniques. The result shows a mean total heterotrophic bacterium count of 5.91 log CFU/ml and a mean microalga count of 4.77 log cells/ml. When crude oil and polycyclic aromatic hydrocarbon (PAH) were used as sole carbon sources, total hydrocarbon utilizing bacteria counts were respectively estimated at 3.89 and 2.89 log CFU/ml. Phenotypic identification of hydrocarbon utilizing bacteria in the crude oil effluents revealed the presence of two main bacterial genera: Streptococcus and Pseudomonas. Data obtained from this study confirmed the biodegradative abilities of indigenous bacterial species, thus, ultimately resulting in the amelioration of the toxicity associated with the crude oil effluents.


Author(s):  
D. N. Ogbonna ◽  
S. I. Douglas ◽  
V. G. Awari

Many substances known to have toxic properties are regularly introduced into the environment through human activity. These substances which include hydrocarbons range in degree of toxicity and danger to human health. Frequent oil spills incidents have become a problem to ecological protection efforts. Conventional methods to remove, reduce or mitigate toxic substances introduced into soil via anthropogenic activities suffer setbacks due to the level of risk involved but bioremediation offers an alternative method to detoxify contaminants especially if the soil conditions are amended with organic nutrients or growth enhancing co-substrates. This study was therefore aimed characterizing hydrocarbon utilizing microorganisms associated with crude oil contaminated soils. Soils were obtained from the Rivers State University Agricultural farm contaminated deliberately with crude oil and allowed for 21 days to mimic the natural polluted soil. Sample collection and analyses were carried out according to standard microbiological procedures while characterization of the isolates was done using genomic studies. The results of microbial counts obtained from the soil samples for total heterotrophic bacteria ranged from 2.10 x108 to 2.58 x108 cfu/g, Total heterotrophic fungi had 1.6 x105 to 2.0 x105 cfu/g while the hydrocarbon utilizing bacteria ranged from 8.0 x103 to 5.0 x104 cfu/g and total hydrocarbon utilizing fungi ranged from 9.0 x103 to 7.0 x104 cfu/g in the contaminated soil. Five hydrocarbon utilizing bacterial species were identified as Staphylococcus saprophyticus, Bacillus amyloliquefaciens, Pseudomonas aeruginosa, Comamonas testosteroni and Chryseobacterium cucumeris while five hydrocarbon utilizing fungal species were identified as Penicillium citrinum, Penicillium brocae, Fusarium solani, Kodamaea ohmeri and Lentinus squarrosulus. Bacillus and Penicillium species were predominantly isolated from the soil. This may be due to the ability of the organisms to produce spores, which may shield them from the toxic effects of the hydrocarbons. Since these organisms are able to utilize crude oil as their sole carbon source. Hence, can be used for bioremediation of crude oil polluted environment.


2001 ◽  
Vol 28 (1) ◽  
pp. 158-174 ◽  
Author(s):  
M Gamache ◽  
J F Blais ◽  
R D Tyagi ◽  
N Meunier

Until now, some microbiological studies have made it possible to highlight the role and identification of certain chimiolithotroph microorganisms directly involved in the simultaneous sewage sludge digestion and metal leaching (SSDML process). Moreover, some other studies have established the performance of the SSDML process for the destruction of pathogens. However, until now no study has been carried out to define the equilibrium of the heterotrophic microbial populations during the sludge acidification occurring during the operation of the SSDML process. Hence, the SSDML process was studied in batch and continuous mode employing different types of sludges (Communauté Urbaine de Québec (CUQ)-East primary sludge, Beauceville secondary sludge, and mixture of primary and secondary Valcartier sludge). The equilibrium of heterotrophic microbial populations was investigated along with sludge acidification and solids degradation. The results obtained during the SSDML process on different sludges showed a decrease followed by the destruction of a majority of heterotrophic bacterial species found in fresh sludges. The isolation and characterization of different microbial species showed that only two types of microorganisms persisted: the yeast Blastoschizomyces capitatus and an unidentified fungus.Key words: sewage sludge, simultanous bioleaching and digestion, heterotrophic bacteria, Blastoschizomyces capitatus, fungi.


Certain microorganisms especially bacteria and fungi are able to use xenobiotic organic compounds as their carbon and nitrogen source for metabolism. Flower farms around lake Naivasha basin uses several agrochemicals especially pesticides to control pests and improve flower production. The aim of this study was to isolate and characterize morphologically and biochemically the main bacterial species that are able to grow and tolerate the pesticide contaminated farm soils. Soil samples were collected from randomly selected five greenhouses from each five flower farms namely Crescent, Elsamere, Karuturi, Malewa and Sewage farms around Lake Naivasha basin. The collected samples were processed for bacterial isolation using the nutrient agar, mac’ Conkey agar, blood agar, Luria-Bertani and Minimum Salt Media nutrient media. The conventional methods of swabbing and streaking were used. Pure colonies of isolates organisms were identified and characterized using standard microbiological technique. Morphological, cultural and biochemical characterization of bacterial species isolated from the flower farm soil samples identified mainly Pseudomonas auriginosa, Escherichia coli, Rhodococcus erythropolis and Bacillus subtilis species. Bacterial growth in pesticide consortia was quantified by monitoring colony growth of the species in liquid culture over time. The viable cell counts were determined turbidimetrically at O.D696nm. All the isolated bacterial species were able to grow in flower farm soil contaminated with organochloride and organophosphate pesticide residues. B. subtilis recorded the highest growth at 1.77±0.07 O.D696nm in pesticide mixture consortia. There was lower growth in organochloride pesticide consortia as compared to organophosphate pesticide consortia.


Author(s):  
H. D. Nyarko ◽  
G. C. Okpokwasili ◽  
O. F. Joel ◽  
I. A. K. Galyuon

Aims: The study aimed at the quantification, isolation and characterization of hydrocarbon degrading bacteria in oil-contaminated and pristine soils. Methodology: Soil samples from petroleum hydrocarbon polluted sites at auto-mechanic workshops, a mechanic village, as well as pristine (control) soils, comprising of 14 sampling locations within Cape Coast Metropolis in the Central Region of Ghana were collected using standard sampling techniques. Collected soil samples were treated and cultured while enumerations, isolations and characterization of carbonoclastic bacteria associated were evaluated. Results: Bacterial populations isolated from hydrocarbon-polluted sites had higher aerobic counts ranging from 7.24-8.02 log10 cfu/g of soil when compared with the pristine sites (from 6.79-7.61 log10 cfu/g of soil). Also, soil samples from the mechanic village (8.76 to 7.48 log10 cfu/g of soil) recorded more bacterial counts than those from the mechanic garages (8.02 to 7.24 log10 cfu/g of soil). The calculated percentage profiles of all the hydrocarbon utilizing bacteria in the total culturable heterotrophic bacteria were low throughout the study, even though the percentage scores were all above 50%. A total of 19 hydrocarbon degraders were isolated. The isolates identified belong to the genera Pseudomonas, Proteus, Bacillus and Enterobacter. Conclusion: The outcome of the study based on the bacteria populations, identification profiles, coupled with their survival and multiplications in designated medium amended with crude oil as the carbon and energy sources, suggest their petroleum hydrocarbon degrading capabilities, hence may be used in bioremediation applications.


Author(s):  
Renner Renner Nrior ◽  
Augusta Ogechi Inweregbu

Aim: To investigate bioremediation potentiality of Pseudomonas aeruginosa KX828570 on crude oil Polluted Marshland and Terrestrial Soil treated with oil spill dispersant Study Design: The study employs experimental design, statistical analysis of data and interpretation. Place and Duration of the Study: Soil samples were collected from K-Dere, Gokana L.G.A, and were transported to the Microbiology Laboratory of Rivers State University, Port Harcourt, Nigeria for analyses while Oil spill dispersant (OSD/LT and OSD/Seacare) were from Barker and Hughes Nig Ltd (formally mil park Nigeria limited), all in Rivers state, Nigeria. This investigation study lasted for 28 days and sampling was done every 7day period. Methodology: Eight experimental set up were carried out using Pseudomonas aeruginosa KX828570 as the bio-augmenting organism in terrestrial and marshland soil contaminated with two Oil spill dispersant (OSD/LT and OSD/Seacare) separately. Controls for the two soil types were made without organism and treatment. Its bioremediation potential on the pollutants and two types of soil were monitored for 28 days. The setup was tilled twice a week to provide moisture and more oxygen for the organisms to thrive. Analysis of pH, Temperature, Moisture content, Total Hydrocarbon Content, Total Heterotrophic Bacteria, Dispersant Utilizing Bacteria was carried out at weekly intervals. Results: The pH of both soils ranged from 5.75 to 7.37; temperature from 270C to 340C; moisture content 03 to 0.6 across the soil samples. Total Hydrocarbon Content (THC) showed a steady decline from day 1 – 28. The percentage (%) bioremediation rates of polluted soils were as follows: Terrestrial soil+OSD/Seacare+Pseudomonas aeruginosa KX828570 69.4% > Terrestrial soil+OSD/LT+ Pseudomonas aeruginosa KX828570 65.9% > Marshland soil+OSD/Seacare+ Pseudomonas aeruginosa KX828570 52.6% > Marshland soil+OSD/Seacare+ Pseudomonas aeruginosa KX828570 47.6%. Dispersant utilizing bacterial count in marshland and terrestrial soil ranged from 6.013 to 7.338 log10Cfu/g and 6.045 to 7.301 log10Cfu/g respectively from Day 1 to 28. Conclusion:  From the investigation, it revealed that Oil spill dispersants are more degradable in terrestrial soil than marshland soil. OSD/Seacare is more biodegradable than OSD/LT in both terrestrial and marshland soil augmented with Pseudomonas aeruginosa KX828570. Thus, Pseudomonas aeruginosa KX828570 have been found to be a potential bioremediation agent in oil spill dispersant polluted marshland and terrestrial soil.


2021 ◽  
Vol 4 (2) ◽  
pp. 35-38
Author(s):  
Hussaini Shettima ◽  
Ibrahim Alkali Allamin ◽  
Nasir Halima ◽  
Haruna Yahya Ismail ◽  
Yusuf Musa

The study was conducted to determine the distribution of hydrocarbon utilizing bacteria in spent engine oil (SEO) contaminated soil. Five mechanical workshops within Maiduguri Metropolis. Five bulk soil sample comprising of one each of the five sites; the sites are Leventis Area on Kashim Ibrahim Way, Damboa Raod, Boiler (commonly called Bola) near Maiduguri Monday Market, Ngomari, on Kano Road, and 1000 Housing Estate, also on Kano Road. Nutrient agar was used, or isolation and enumeration total heterotrophic bacteria and Mineral salt agar was used for the isolation and enumeration of hydrocarbon utilizing bacteria. The result shows THB (92.0x104) at Damboa road (DR) to as higher and (78.8x104) at Bola area (BL) while HUB (2.0x104) at Damboa road (DR) and as high as (9.3x104) at Bola area (BL). The bacterial species isolated were species of Bacillus licheniformis, Bacillus subtilis, Bacillus coagulans, Bacillus alvei, Bacillus cereus, Bacillus lentus other are Pseudomonas aeruginosa, Klebsiella pneumonia. Bacillus licheniformis, and Bacillus subtilis are the most occurring bacterial isolates identified. The ability of those bacterial isolate to degrade hydrocarbon buoyantly will help in remediation of oil polluted environments.


2014 ◽  
Vol 3 (2) ◽  
pp. 36-47 ◽  
Author(s):  
IA Allamin ◽  
UJJ Ijah ◽  
HY Ismail ◽  
ML Riskuwa

Soil samples were collected from five sites covering petroleum exploration station in Kukawa, Kukawa Local Government Area of Borno State, Nigeria between October, 2012 and February, 2013 at two different depths (0-10cm and 10-20cm) to enumerate and identify hydrocarbon degrading bacteria in the soil. Total aerobic heterotrophic bacteria (TAHB) were enumerated on Nutrient agar (NA), and Hydrocarbon utilizing bacteria (HUB) enumerated on Oil agar (OA). The bacterial isolates were identified using morphological and biochemical tests. It was observed that the microorganisms (TAHB, and HUB) were more densely populated at 10cm depth. (TAHB: 5.3×108 - 11.4×108cfu/g, and HUB: 2.4×105 - 5.3×105 cfu/g, than at 20 cm depth (TAHB: 3.0×108 - 5.7×108 cfu/g, and HUB: 2.1×105 - 4.8×105 cfu/g). The HUB was identified as species of Bacillus, Pseudomonas, Klebsiella, Lactobacillus, Micrococcus, Corynebacterium, and Actinomyces. Bacillus, and Pseudomonas species were more constantly isolated than other isolates and they constitute 100% of total bacterial isolates. The potential of hydrocarbon utilizing bacteria isolated to degrade hydrocarbon was studied. Nineteen (19) bacterial species was screened, Bacillus subtilis, Pseudomonas aeruginosa, Bacillus cereus, Klebsiella pneumoniae, Micrococcus leteus,and Lactobacillus casei, utilized and degrade crude oil at considerably high rates after 21 days of incubation. The degradation efficiency was confirmed by GC-MS analysis, which indicated that the bacterial isolates utilized most of the crude oil components particularly straight chain alkanes and cycloalkanes DOI: http://dx.doi.org/10.3126/ije.v3i2.10503 International Journal of the Environment Vol.3(2) 2014: 36-47


Author(s):  
David N. Ogbonna ◽  
Renner R. Nrior ◽  
Festus E. Ezinwo

Aim: To assess the Bioremediation efficiency of Bacillus amyloliquefaciens and Pseudomonas aeruginosa strain CL 9 with nutrient amendment using bio-stimulating agents such as Fish waste and Goat manure on crude oil polluted soils in Rivers State, Nigeria. Study Design: The study employs experimental design, statistical analysis of the data and interpretation. Place and Duration of Study: A portion of Rivers State University demonstration farmland in Nkpolu-Oroworukwo, Mile 3 Diobu area of Port Harcourt, Rivers State was used for this study. The piece of land is situated at Longitude 4°48’18.50’’N and Latitude 6o58’39.12’’E measuring 5.4864 m x 5.1816 m with a total area of 28.4283 m2. Bioremediation monitoring lasted for 56 days, analysis carried out weekly (per 7 days interval). Methodology: Seven (7) experimental plots were employed using a Randomized Block Design each having dimensions of 100 x 50 x 20 cm (Length x Breadth x Height) were formed and mapped out on agricultural soil and left fallow for 6 days before contamination on the seventh day; after which it was allowed for 21 days for proper contamination and exposure to natural environmental factors to mimic crude oil spill site. Thereafter bio stimulating agents usually referred to as nutrient amendment organics in this study (fish waste and goat manure) and bio-augmenting microorganisms were applied. Soil profile before and after contamination was assayed while parameters like Nitrate, Sulphate, Phosphate, Total Organic Carbon (TOC) and Total Petroleum Hydrocarbon (TPH), were monitored throughout the experimental period. Microbial analyses such as Total Heterotrophic Bacteria (THB), Total Heterotrophic Fungi (THF), Hydrocarbon Utilizing Bacteria (HUB) and Hydrocarbon Utilizing Fungi (HUF) were recorded. Bioremediation efficiency was estimated from percentage (%) reduction of Total Petroleum Hydrocarbon (TPH) from day 1 to the residual hydrocarbon at day 56 of bio augmented/ biostimulation plots with the control. Results: Results revealed amount of remediated hydrocarbon and % Bioremediation efficiency at 56 days in the different treatment plots (initial TPH contamination value of  9296.83  mg/kg) in a decreasing order as follows: PS+Bac+Pse+GF+FW (8032.825 mg/kg; 86.40%) >PS+GF+FW (6867.825 mg/kg; 73.87%) >PS+Bac+Pse (6587.825mg/kg; 70.86%) >PS+FW (6441.825mg/kg; 69.29%) >PS+GF (5909.825 mg/kg; 63.57%) >CTRL 2 (Polluted soil without amendment) (3604.825mg/kg; 38.78%). Microbiological results showed increased colonial values with increase time exposure. The results observed on day 56 indicate that Polluted soil + Bacillus + Pseudomonas (10.11 Log10 CFU/g) > Polluted soil but un-amended soil (8.76 Log10 CFU/g) > unpolluted soil (8.68 Log10 CFU/g). Comparatively, Polluted soil +Bacillus + Pseudomonas expressed higher heterotrophic bacteria of 9.77 and 9.67 Log10 CFU/g while fungal counts recorded 6.04 and 6.82 Log10 CFU/g. Conclusion: Study showed that bioremediation of crude oil-polluted soils with bacteria singly is less effective but a combination with other organic nutrients is a better palliative measure. Therefore, amendment with organic nutrients like Goat manure and Fish wastes is recommended for crude oil polluted soils due to its high nutrient content as substrates for biostimulation of indigenous and augmenting biodegrading microbes. This process could be a source of enhanced natural attenuation of oil-contaminated environments in Nigeria.


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