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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.
Frutos FJG, Escolano O, García S, Babín M, Fernández MD. Bioventing remediation and ecotoxicity evaluation of phenanthrene-contaminated soil. Journal of Hazardous Materials. 2010;183(1–3):806–813.
Amro MM. Treatment techniques of oil-contaminated soil and water aquifers. International Conf. on Water Resources & Arid Environment. 2004;1–11.
Bidoia ED, Montagnolli RN, Lopes PRM. Microbial biodegradation potential of hydrocarbons evaluated by colorimetric technique: A case study. Appl Microbiol Biotechnol. 2010;7:1277–1288.
Djelal H, Amrane A. Biodegradation by bioaugmentation of dairy wastewater by fungal consortium on a bioreactor lab-scale and on a pilot-scale. 2013;1.
Okerentugba PO, Orji FA, Ibiene AA, Elemo GN. Spent mushroom compost for bioremediation of petroleum hydrocarbon polluted soil: A review. Global Advanced Research Journal of Environmental Science and Toxicology. 2015;4(1):001–007.
Ahlawat OP, Gupta P, Kumar S, Sharma DK, Ahlawat K. Bioremediation of fungicides by spent mushroom substrate and its associated microflora. Indian Journal of Microbiology. 2010;50(4):390–395.
Peuke AD, Rennenberg H. Phytoremediation: Molecular biology, requirements for application, environmental protection, public attention and feasibility. EMBO Reports. 2005; 6(6):497–501.
Salt DE, Blayloc M, Kumar NP, Dushenkov V, Ensley BD, Chet I. Phytoremediation: A novel strategy for the removal of toxic metals from the environment using plants. Bio/Technology. 1995;13(5):468.
USEPA. United states environmental protection agency. Exposure Factors Handbook; 2011.
Bockelmann A, Zamfirescu D, Ptak T, Grathwohl P, Teutsch G. Quantification of mass fluxes and natural attenuation rates at an industrial site with a limited monitoring network: A case study. Journal of Contaminant Hydrology. 2003;60(1–2):97–121.
Toogood TA. Effect of oil spill on the physical properties of soils. In: J.A. Toogood (ed). The reclamation of agricultural soil after oil spills. Part 1 Research, Alberta Institute of Pedology, Canada, Publication No. M-11-77. 1977; 108-121.
APHA. Standard methods for the examination of water and waste water. 20th ed. APHA-AWWA-WPCF. Washington DC; 1998.
Nrior RR, Mene GB. Assessment of bioaugmentation efficiency of penicillium chrysogenum and aspergillus nidulans bioremediation of crude oil spill soil. Journal of Environmental Science, Toxicology and Food Technology. 2017; 11(8):01-09.
Menkit MC, Amechi AK. Evaluation of first and second order degradation rates and biological half-lives in crude oil polluted soil. Asian Journal of Biotechnology and Genetic Engineering. 2019;2(1):1-11.
Ogbonna DN, Nrior RR, Ezinwo FE. Bioremediation efficiency of bacillus amyloliquefaciens and pseudomonas aeruginosa with the nutrient amendment on crude oil polluted the soil. Microbiology Research Journal International. 2019;1–13. Available:https://doi.org/10.9734/mrji/2019/v29i530175
Nrior RR, Odokuma LO. Ultimate biodegradability potential of trichloroethylene (TCE) used as degreaser in marine, brackish and fresh water. Journal of Environmental Sciences, Toxicology and Food Technology. 2015;9:80-89.
Nrior RR, Echezolom C. Assessment of percentage bioremediation of Petroleum Hydrocarbon polluted soil with biostimulating agents. Journal of Current Studies in Comparative Education, Science and Technology. 2017;3(1):203-215.
Chikere CB, Okpokwasili GC, Chikere BO. Bacterial diversity in a tropical crude oil polluted soil undergoing bioremediation. African Journal of Biotechnology. 2009; 8(11):2535-2540.
Ollivier B, Magot M. Petroleum Microbiology. Washington, DC: ASM: 12/08/2017; Heat of Combustion of Fuels. 2005;1.
Kidd S, Halliday C, Alexiou H, Ellis D. Description of medical fungi (3rd ed). Adelaide, Australia. 2016;232-235.
Cheesbrough M. District Laboratory Practice in Tropical Countries. 2006;2-5
Jat ML, Bijay S, Stirling CM, Jat HS, Tetarwal JP, Jat RK. Chapter Four—Soil Processes and Wheat Cropping Under Emerging Climate Change Scenarios in South Asia. In D. L. Sparks (Ed.), Advances in Agronomy. 2018;148:111–171. Academic Press. 2017.11.006 Available:https://doi.org/10.1016/bs.agron
Harms H. Bioavailability and Bioaccessibility as Key Factors in Bioremediation. In M. Moo-Young (Ed.), Comprehensive Biotechnology (Second Edition). 2011;83–94. Academic Press. Available:https://doi.org/10.1016/B978-0-08-088504-9.00367-6
Liu GM, Yang JS, Yao RJ. Electrical Conductivity in Soil Extracts: Chemical Factors and Their Intensity1 1Project supported by the National Basic Research Program of China (No. 2005CB121108), the National Natural Science Foundation of China (No. 40371058), and the National High Technology Research and Development Program of China (863 Program) (No. 2002AA2Z4061). Pedosphere. 2006;16(1):100–107. Available:https://doi.org/10.1016/S1002-0160(06)60031-3
Dawoodi V, Madani M, Tahmourespour A, Golshani Z. The Study of Heterotrophic and Crude Oil-utilizing Soil Fungi in Crude OilContaminated Regions. Journal of Bioremediation and Biodegradation. 2015;6(2):1–5. Available:https://doi.org/10.4172/2155-6199.1000270
Ayuba SA, John C, Obasi MO. Effects of organic manure on soil chemical properties and yield of ginger—Research note. Nigerian Journal of Soil Science. 2005;15:136–138. Available:https://doi.org/10.4314/njss.v15i1.37461
Nrior RR, Jirigwa CC. Comparative bioremediation potential of mucor racemosus and paecilomyces variotii on crude oil spill site in Gio Tai, Ogoni land. IOSR Journal of Environmental Science, Toxicology and Food Technology (IOSR-JESTFT). 2017;11(10):49-57.
Oa E, Fi A. Bioremediation of petroleum hydrocarbons from crude oil-contaminated soil with the earthworm: Hyperiodrilus africanus. 3 Biotech. 2015;5(6):957–965. Available:https://doi.org/10.1007/s13205-015-0298-1
Neina D. The role of soil pH in plant nutrition and soil remediation [Review Article]. Applied and Environmental Soil Science; Hindawi. 2019;1-3. Available:https://doi.org/10.1155/2019/5794869
Benyahia F, Embaby AS. Bioremediation of crude oil contaminated desert soil: Effect of biostimulation, bioaugmentation and bioavailability in biopile treatment systems. International Journal of Environmental Research and Public Health. 2016;13(2). Available:https://doi.org/10.3390/ijerph13020219
Ebuehi OAT, Abibo IB, Shekwolo PD, Sigismund KI, Adoki A, Okoro IC. Remediation of crude oil contaminated soil by enhanced natural attenuation technique. Journal of Applied Sciences and Environmental Management. 2005; 9(1):1. Available:https://doi.org/10.4314/jasem.v9i1.17265
Dalefield R, Chapter 18. Industrial and occupational toxicants. In R. Dalefield (Ed.), Veterinary Toxicology for Australia and New Zealand, Elsevier. 2017;333–341. Available:https://doi.org/10.1016/B978 0-12-420227-6.00017-7
Wang Y, Tam NFY. Chapter 16—Microbial Remediation of Organic Pollutants. In C. Sheppard (Ed.), World Seas: An Environmental Evaluation (Second Edition) 2019;283–303. Academic Press. Available:https://doi.org/10.1016/B978-0-12-805052-1.00016-4