scholarly journals The Ex-Situ Bioremediation Kinetics of Raw and Treated Crude Oil Polluted Soil Using Aspergillus Niger and Pseudomonas Aeruginosa

Author(s):  
Modupe Ojewumi ◽  
Valentina Ejemen

The study was done to investigate the kinetics of first order bioremediation. The effectiveness of remediating soils polluted with raw crude oil and treated crude oil using Aspergillus niger (fungi) and Pseudomonas aeruginosa (bacteria) were investigated. Eight systems of 500g soil sample were polluted with both raw and treated crude oil. Four systems were polluted with 40g treated crude oil while the other remaining four systems were polluted with 40g raw crude oil. Two systems with raw crude and treated crude were left as control (RCC and TCC). Raw crude samples were treated with Aspergillus niger only (RCA) and Pseudomonas aeruginosa (RCP) while treated crude samples were also treated with same (TCA) and (TCP) only. The last two systems were treated with both Pseudomonas aeruginosa and Aspergillus niger (RCAP and TCAP). The first order bioremediation kinetics and biostimulant efficiency for these systems were studied by monitoring Total Petroleum Hydrocarbon (TPH). At the end of the bioremediation period, the results obtained showed that treated crude oil polluted soil generally remediated faster and better than raw crude oil polluted soil. The highest level of bioremediation occurred in systems amended with both Pseudomonas aeruginosa and Aspergillus niger which had about 98% TPH decrease.

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

Aim: To assess the Mycoremediation potential of Mucor racemosus and Aspergillus niger in open field crude oil contaminated soils in Rivers State, Nigeria.  Study Design: The study employs experimental design, statistical analysis of the data and interpretation. Place and Duration of Study: Rivers State University demonstration farmland in Nkpolu-Oroworukwo, Mile 3 Diobu area of Port Harcourt, 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 square meter. Mycoremediation process monitoring lasted for 56 days, analyses were carried out weekly at 7 days’ interval. Methodology: Five (5) experimental plots were employed using a Randomized Block Design each having dimensions of 100 x 50 x 30 cm (Length x Breadth x Height) and were formed and mapped out on agricultural soil, each plot was contaminated with 22122.25g of Crude Oil except Control 1 and left fallow for 6 days after contamination for proper contamination and exposure to natural environmental factors to mimic crude oil spill site. On the seventh day bio-augmentation process commenced using two (2) fungal isolates namely Aspergillus niger [Asp] and Mucor rasemosus [Muc]). Two (2) control plots (P1: Uncontaminated and unamended soil - CTRL 1 US) and P2: Crude Oil contaminated but unamended soil - CTRL 2 CS); P3 = P5 were contaminated and amended/bioaugmented (P3: CS+Asp, P4: CS+Muc, P5: CS+Asp+Muc respectively. Soil profile before and after contamination was assayed while parameters like Temperature, pH, Nitrogen, Phosphorus, Potassium and Total Petroleum Hydrocarbon (TPH) contents 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- augmentation/ biostimulation plots with the control. Results: Results revealed actual amount of remediated hydrocarbon and % Bioremediation Efficiency at 56 days in the different treatment plots (initial TPH contamination value of 8729.00mg/kg) in a decreasing order as follows: CS+Muc (8599.19mg/kg; 33.66%) > CS+Asp+Muc (8357.31mg/kg; 33.04%) > CS+Asp (8341.58mg/kg; 32.98%) > CTRL 2 -CS (Polluted soil without amendment) (81.06mg/kg; 0.32%). Microbiological results After fifty-six (56) days of bioremediation monitoring; %HUB were as follows; CS+Asp+Muc (45.30%) > CS+Asp (40.32%) > CS+Muc (35.01%) > CTRL 2 –CS (30.43%) > CTRL 1 – US (0%). These results indicate that the presence of the contaminated crude oil stimulated and sustained the growth of Hydrocarbon Utilizing Bacteria (HUB) in the contaminated plots (P2 - P3); more so, the higher growth in the enhanced bio-augmented plots (P3 – P5) shows the positive impact of fungal bio-augmentation in bioremediation of crude oil polluted soil. It was further observed that treatment plots with higher HUB or HUF had higher percentage (%) bioremediation efficiency; that is, the higher the sustained HUB and HUF population, the higher the %Bioremediation process. Hydrocarbon Utilizing Bacteria (Log10 CFU/g): CS+Asp (4.20) (Day 35) > CS+Muc+Asp (4.18) (Day 35) > CS+Muc (4.08) (Day 28) > CTRL 2 – CS (3.95) (Day 21) > CTRL 1 – US (3.78) (Day 35). (Fig. 3). Hydrocarbon Utilizing Fungi (Log10 CFU/g): CS+Asp (4.68) (Day 35) > CS+Muc+Asp (4.58) (Day 35) > CS+Muc (4.48) (Day 35) > CTRL 2 – CS (4.23) (Day 21) > CTRL 1 – US (2.85) (Day 42). Conclusion: Study showed that bioremediation of crude oil-contaminated soils with Bioaugmenting fungus singly may be more effective than combination with others depending on the type of substrate used, nature of the hydrocarbon utilizing organism and environmental conditions prevalent as seen in Mucor racemosus having higher bioremediation potential than when combined with Aspergillus niger. Notably, Hydrocarbon Utlilizing Bacteria (HUB) and Hydrocarbon Utilizing Fungi (HUF) which are the key players in Bioremediation has its peak count value on Day 35, this confers that nutrient renewal on bioremediation site should be at interval of 35 days for continuous effective bioremediation of hydrocarbon pollutants. It is therefore recommended that single microbes of high bioremediation potential could be used since its more effective than consortium of many hydrocarbon utilizing microbes. Also, nutrient or bio-augmenting microbes’ renewal on bioremediation site should be at an interval of 35 days for continuous effective bioremediation of hydrocarbon pollutants.


2021 ◽  
Author(s):  
Bobby Chettri ◽  
Ningombam Anjana Singha ◽  
Arvind Kumar Singh

Abstract We report kinetics of Assam crude oil degradation by Pseudomonas aeruginosa AKS1 and Bacillus sp. AKS2, both isolated from Assam refinery sediments. The isolates exhibited appreciable degrees of hydrophobicity, emulsification index and biosurfactant production. Crude oil degradation efficiency of isolates was assessed in (1) liquid medium amended with 1% v/v crude oil and (2) microcosm sediments (125 mg crude oil/ 10 g sand). In liquid culture, the biodegradation rate (k) and half-life (t1/2) values were found to be 0.0383 day -1 and 18.09 days for P. aeruginosa AKS1, and 0.0204 day -1 and 33.97 days in case of Bacillus sp. AKS2. In microcosm sand sediments, the estimated biodegradation rate (k) and half-life (t 1/2) values were 0.0138 day -1 and 50 days for P. aeruginosa AKS1, and 0.0113 day -1 and 61.34 days in case of Bacillus sp. AKS2. The level of nutrient treatment in microcosm sand sediment was 125 µg N & 62.5 µg P/g sediment in case of P. aeruginosa AKS1 and 375 µg N & 37.5 µg P/g sediment in case of Bacillus sp. AKS2. In microcosms without inorganic nutrients, biodegradation rate (k) and half-life (t1/2) values were found to be 0.0069 day -1 and 100 days for P. aeruginosa AKS1 and for Bacillus sp. AKS2, the respective values were found to be 0.0046 day -1 and 150.68 days. Our data provides important information for predictive hydrocarbon degradation in liquid medium and contaminated sediments.


Author(s):  
J. O. Dasetima-Altraide ◽  
D. N. Ogbonna ◽  
T. K. S. Abam ◽  
A. E. Gobo

Aim: To assess the Physicochemical indices of Phytoremediated Crude Oil polluted amended soil using grass plant Cyperus esculentus (Cyp) and Phyllanthus amarus (Phy). Study Design: The study employs experimental design, statistical analysis of the data and interpretation. Place and Duration of Study: Rivers State University demonstration farmland in Nkpolu- Oroworukwo, Mile 3 Diobu area of Port Harcourt, 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 square meter. Phytoremediation process monitoring lasted for 240 days; analyses were carried out monthly at 30 days’ interval. Methodology: The study was carried out on Crude Oil Polluted soil (PS) amended with bio-nutrient supplements (Spent Mushroom Substrate (SMS) and selected fungi (Aspergillus niger(AN) andMucor racemosus (MR)) used to stimulate and augment the indigenous microbial population present in a crude oil polluted soil thereby enhancing hydrocarbon reduction in pari per sue with phytoremediation (uptake of Crude oil by test plants) over a period of 240 days. Ten (10) experimental plots (two Control (Unpolluted and polluted soil without amendment) and eight polluted amended/treated plots) employing Randomized Block Design (each having dimensions: 100 x 50 x 30 cm LxBxH); 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 soil crude oil spill site); thereafter nutrients/organics (biostimulating agents) and bioaugmenting organisms were applied. Baseline studies were carried out on soil profile before and after contamination, major parameters monitored and assayed were Total Petroleum Hydrocarbon (TPH) uptake by plant roots and stem, Polycyclic Aromatic Hydrocarbons (PAHs) and TPH reduction in soil. Other physicochemical properties analyzed in the soil from different plots were pH, Electrical Conductivity, Moisture Content, Total Nitrogen, Available Phosphorus, Potassium, Total Organic Carbon, Plant Height, Iron, Lead and Zinc at regular intervals; days 1, 60, 90, 120, 150, 180, 210 & 240. The rate of phytoremediation was estimated from percentage (%) uptake of Total petroleum hydrocarbon (TPH) in plant roots and stem from day 1 -240; while percentage (%) reduction of TPH and PAHs in soil was estimated from day 1 to the residual at day 240. Results: The test plants decreased significant amount of crude oil as revealed in TPH uptake in their roots and Stem. Mean amount and percentage Total Petroleum Hydrocarbon (TPH) uptake by Cyperus esculentus roots and stem were; 152.33±50.34mg/kg, 12.57±4.16% and 201.13±8.80mg/kg, 13.27±0.58% respectively; while that of Phyllanthus amarusroots and stem were 141.50±35.62mg/kg, 11.68±2.94% and 174.44±19.98mg/kg, 11.51±1.32% respectively; revealing higher Uptake of TPH in plant stem than roots. From the initial TPH contamination value of 5503.00mg/kg, it was observed that plots planted with Cyperus esculentus (TPH 5492.75±76.36mg/kg) showed higher reduction of TPH from soil than those planted with Phyllanthus amarus(TPH 5449.72±18.27mg/kg); while PAHs degradation/reduction showed a reverse trend with plots planted with Phyllanthus amarus (PAHs 28.72±2.74mg/kg; 60.46±5.77%) higher than plots planted with Cyperus esculentus s (PAHs 25.77±2.12mg/kg, 54.24±4.47%). Conclusion: Plots planted with Cyperus esculentus showed higher reduction of TPH from soil than those planted with Phyllanthus amarus while PAHs degradation/reduction in plots planted with Phyllanthus amarus was higher than plots planted with Cyperus esculentus. TPH uptake was higher in plant stems than roots; more so, plots amended with nutrient supplements showed significant higher percentage reduction in hydrocarbon in the polluted soil than unamended polluted soil. It is therefore recommended that Cyperus esculentus is a suitable plant species for phytoremediation of crude oil contaminated soil with high TPH value while Phyllanthus amarusis the best option in phytoremediation of polluted soil with high PAHs value, both in combination with bio-nutrient supplement.


2016 ◽  
Vol 216 ◽  
pp. 548-558 ◽  
Author(s):  
Bobby Chettri ◽  
Arghya Mukherjee ◽  
James S. Langpoklakpam ◽  
Dhrubajyoti Chattopadhyay ◽  
Arvind K. Singh

2018 ◽  
Vol 17-18 ◽  
pp. 196-204 ◽  
Author(s):  
Modupe Elizabeth Ojewumi ◽  
Joshua Olusegun Okeniyi ◽  
Elizabeth Toyin Okeniyi ◽  
Jacob Olumuyiwa Ikotun ◽  
Valentina Anenih Ejemen ◽  
...  

Author(s):  
Amirreza Talaiekhozani ◽  
Nematollah Jafarzadeh ◽  
Mohamad Ali Fulazzaky ◽  
Mohammad Reza Talaie ◽  
Masoud Beheshti

2020 ◽  
Vol 22 (2) ◽  
pp. 67-72
Author(s):  
Justyna Miłek

AbstractThe thermal stability of enzyme-based biosensors is crucial in economic feasibility. In this study, thermal deactivation profiles of catalase Aspergillus niger were obtained at different temperatures in the range of 35°C to 70°C. It has been shown that the thermal deactivation of catalase Aspergillus niger follows the first-order model. The half-life time t1/2 of catalase Aspergillus niger at pH 7.0 and the temperature of 35°C and 70°C were 197 h and 1.3 h respectively. Additionally, t1/2 of catalase Aspergillus niger at the temperature of 5°C was calculated 58 months. Thermodynamic parameters the change in enthalpy ΔH*, the change in entropy ΔS* and the change Gibbs free energy ΔG* for the deactivation of catalase at different temperatures in the range of 35°C to 70°C were estimated. Catalase Aspergillus niger is predisposed to be used in biosensors by thermodynamics parameters obtained.


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.


Author(s):  
Mariana MARINESCU ◽  
Anca LACATUSU ◽  
Eugenia GAMENT ◽  
Georgiana PLOPEANU ◽  
Vera CARABULEA

Bioremediation of crude oil contaminated soil is an effective process to clean petroleum pollutants from the environment. Crude oil bioremediation of soils is limited by the bacteria activity in degrading the spills hydrocarbons. Native crude oil degrading bacteria were isolated from different crude oil polluted soils. The isolated bacteria belong to the genera Pseudomonas, Mycobacterium, Arthrobacter and Bacillus. A natural biodegradable product and bacterial inoculum were used for total petroleum hydrocarbon (TPH) removal from an artificial polluted soil. For soil polluted with 5% crude oil, the bacterial top, including those placed in the soil by inoculation was 30 days after impact, respectively 7 days after inoculum application, while in soil polluted with 10% crude oil,  multiplication top of bacteria was observed in the determination made at 45 days after impact and 21 days after inoculum application, showing once again how necessary is for microorganisms habituation and adaptation to environment being a function of pollutant concentration. The microorganisms inoculated showed a slight adaptability in soil polluted with 5% crude oil, but complete inhibition in the first 30 days of experiment at 10% crude oil.


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