scholarly journals Microbial studies of biosurfactant producing bacteria from crude oil contaminated soil

2021 ◽  
Vol 25 (9) ◽  
pp. 1729-1735
Author(s):  
K.I. Ogru ◽  
P.G. Olannye
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacillus Subtilis ◽  
Crude Oil ◽  
Contaminated Soil ◽  
Heterotrophic Bacteria ◽  
Metal Removal ◽  
Soil Samples ◽  
Optimum Growth ◽  
The Mean ◽  
Hydrocarbon Utilizing Bacteria

Soils contaminated with crude oil collected from three sites in Delta State were analyzed using serial dilution and pour plate methods. The physicochemical characteristics of the soil were determined using standard methods. The mean heterotrophic bacteria count and the mean hydrocarbon utilizing bacteria count were also determined. Blood haemolysis, foaming activity, oil spreading techniques and emulsification assay was used to screen the microorganisms for biosurfactant production. The biosurfactant producing bacteria were selected and their effect on metal removal and growth kinetics was also determined. The pH of the contaminated soil samples ranged 4.82 to 5.62. The mineral elements such as potassium (k+) and sodium (Na+) ranged from 0.20 to 0.80 Meq/100g and 0.07 to 0.81 Meq/100g respectively. The heavy metal content such as zinc (Zn2+) and lead (Pb2+) ranged from 10.13 to 19.24 mg/kg and 19.24 to 49.63 mg/kg respectively. Organic carbon and THC ranged from 5.44 to 6.87 % and 2720.00 to 3110.00 mg/kg respectively. The mean heterotrophic bacteria and hydrocarbon utilizing bacteria ranges from 1.4 ± 0.4 x 106 to 1.6 ± 0.7 x 106 and 1.0 ± 0.2 x 106 to 1.2 ± 0.4 × 106 respectively in the soil samples. Bacteria isolated and identified from the three sites include Corynebacterium spp., Bacillus subtilis., Pseudomonas aeruginosa, Micrococcus varians, Streptococcus spp., Klebsiella spp. and Enterobacter spp. with Bacillus subtilis and Pseudomonas aeruginosa having the highest prevalence and among these isolates only Corynebacterium spp., Bacillus subtilis., Pseudomonas aeruginosa and Micrococcus varians showed positive result for the production of biosurfactant. Bacillus subtilis had the highest metal removal capacity of 29.44% of 100 ppm of lead. Corynebacterium spp. and Bacillus subtilis had an optimum growth at pH 8 and 7 respectively while Pseudomonas aeruginosa and Micrococcus varians has an optimum growth at pH.

scholarly journals Percentage Bioremediation Assessment of Spent Mushroom Substrate (SMS) and Mucor racemosus in Hydrocarbon Contaminated Soil

2020 ◽  
pp. 1-21
Author(s):  
D. N. Ogbonna ◽  
S. A. Ngah ◽  
R. N. Okparanma ◽  
O. Ule ◽  
R. R. Nrior
Keyword(s):  
Crude Oil ◽  
Contaminated Soil ◽  
Petroleum Hydrocarbon ◽  
Heterotrophic Bacteria ◽  
Polluted Soil ◽  
Total Petroleum Hydrocarbon ◽  
Spent Mushroom Substrate ◽  
Mucor Racemosus ◽  
Rivers State ◽  
Hydrocarbon Utilizing Bacteria

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.

scholarly journals Characterization of Hydrocarbon Utilizing Bacteria and Fungi Associated with Crude Oil Contaminated Soil

2020 ◽  
pp. 54-69
Author(s):  
D. N. Ogbonna ◽  
S. I. Douglas ◽  
V. G. Awari
Keyword(s):  
Crude Oil ◽  
Contaminated Soil ◽  
Contaminated Soils ◽  
Heterotrophic Bacteria ◽  
Anthropogenic Activities ◽  
Bacterial Species ◽  
Soil Conditions ◽  
Penicillium Species ◽  
Hydrocarbon Utilizing Bacteria

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.

scholarly journals Bioremediation Potential of Pseudomonas aeruginosa KX828570 on Crude Oil Spill Polluted Marshland and Terrestrial Soil Treated with Oil Spill Dispersant

2019 ◽  
pp. 1-15
Author(s):  
Renner Renner Nrior ◽  
Augusta Ogechi Inweregbu
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Moisture Content ◽  
Crude Oil ◽  
Oil Spill ◽  
Heterotrophic Bacteria ◽  
Soil Samples ◽  
Total Hydrocarbon ◽  
Hydrocarbon Content ◽  
Rivers State ◽  
Total Hydrocarbon Content

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.

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
Keyword(s):  
Crude Oil ◽  
Heterotrophic Bacteria ◽  
Bacterial Species ◽  
Polluted Soil ◽  
Soil Samples ◽  
Isolation And Characterization ◽  
Significant Difference ◽  
Rivers State ◽  
Hydrocarbon Utilizing Bacteria

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.

Assessment of bacterial diversity associated with crude oil-contaminated soil samples from Assam

2017 ◽  
Vol 14 (10) ◽  
pp. 2155-2172 ◽  
Author(s):  
R. Baruah ◽  
S. K. Mishra ◽  
D. J. Kalita ◽  
Y. Silla ◽  
P. S. Chauhan ◽  
...  
Keyword(s):  
Crude Oil ◽  
Bacterial Diversity ◽  
Contaminated Soil ◽  
Soil Samples

scholarly journals Isolation and Characterization of Crude Oil Degrading Bacteria in Association with Microalgae in Saver Pit from Egbaoma Flow Station, Niger Delta, Nigeria

2020 ◽  
Vol 2 (2) ◽  
pp. 12-16
Author(s):  
Obhioze Augustine Akpoka
Keyword(s):  
Crude Oil ◽  
Heterotrophic Bacteria ◽  
Oil Pollution ◽  
Heterotrophic Bacterium ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Sole Source ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
Hydrocarbon Utilizing Bacteria

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.

scholarly journals Effect of Aerating Duration on Hydrocarbon Biodegradation in a Simulated Crude-Oil Polluted Aquatic Environment Undergoing Bioremediation

2021 ◽  
pp. 29-39
Author(s):  
Ewoh, Prosperity C. ◽  
Peekate, Lekiah P.
Keyword(s):  
Crude Oil ◽  
Aquatic Environment ◽  
Heterotrophic Bacteria ◽  
Research Work ◽  
Hydrocarbon Biodegradation ◽  
Slight Reduction ◽  
Biodegradation Rate ◽  
Reaction Equation ◽  
First Order ◽  
Hydrocarbon Utilizing Bacteria

The aim of this research work was to determine the aerating duration that would be effective in enhancing hydrocarbon biodegradation rate during bioremediation of crude-oil polluted river. Sediment and river-water were placed in four glass troughs labeled CT (control), A, B, and C. The setups were polluted with crude-oil, and allowed undisturbed for 2 weeks. Subsequently, accessible crude-oil on the surface was removed; bacteria and nutrients were then added. Air was bubbled for 3 hours into setups A, B, and C, at daily, 3 days, and 7 days interval respectively. Aeration was not applied to setup CT. On day 1, 7, 14, and 21, hydrocarbon concentration was determined; populations of total heterotrophic bacteria (THB) and hydrocarbon-utilizing bacteria (HUB) were also determined. The time it will take for hydrocarbons in the setups to biodegraded “completely” was calculated using first-order reaction equation. The results obtained showed that 71.43, 86.39, 83.17, and 15.42 % hydrocarbon degradation were obtained in setup A, B, C, and CT respectively. The time it will take for hydrocarbons in the setups to biodegrade “completely” were 129, 89, 101, and 1079 days for A, B, C, and CT respectively. There was slight reduction in population of HUB in setup CT, fairly stable population in setup A, and increase in population of HUB in setups B and C. It is concluded that aerating crude-oil polluted aquatic environment for 3 hours at 3 days interval will be more effective in enhancing hydrocarbon biodegradation rate during bioremediation.

scholarly journals INDUCED DEGRADATION OF CRUDE OIL POLLUTED SOIL BY MICROBIAL AUGMENTATION

2017 ◽  
Vol 23 (4) ◽  
pp. 37-44
Author(s):  
SYLVESTER UWADIAE ◽  
EMIKE OMOAYENA
Keyword(s):  
Crude Oil ◽  
Soil Sample ◽  
Contaminated Soil ◽  
Polluted Soil ◽  
Soil Samples ◽  
The Other ◽  
Pseudomonas Sp ◽  
Hydrocarbon Content ◽  
Percentage Removal

This study was aimed at assessing the effectiveness of indigenous microbes for remediation of hydrocarbon contaminated soil by first increasing the population of the indigenous microbes via bioaugmentation. Soil samples were treated using a consortium of bacteria: Bacillus substilis and Pseudomonas sp. which were isolated and cultured from the contaminated soil. The non-bacteria injected soil sample had the highest hydrocarbon content (THC) of 271.021 in comparison with the other soil samples. The THC percentage removal of B1 (96.885 %), B2 (97.562 %), B3 (98.835 %), B4 (99.594 %) and B5 (99.540 %) were higher than that of the control; indicating that biodegradation actually took place.

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