Cover Image

Biostimulation of Immobilized Microbes in a Fixed-Bed Bioreactor towards Hydrocarbon-Contaminated Water Remediation Using Compost Tea and Egg Shell Powder

Samuel Antwi-Akomeah, Bernard Fei-Baffoe, Ebenezer J.D. Belford, Paul Osei-Fosu


The need to satisfy the ever increasing petroleum demand coupled with the availability of enough crude oil reserves to meet demand for several decades to come makes petroleum contamination a globally important subject. Bioremediation has emerged as an efficient treatment option for the remediation of organic contaminants including petroleum owing to the failure of physico-chemical treatment options to address petroleum contamination. The current study targeted the application of compost tea (CT) and egg shell powder (ESP) solution at various levels as nutrient amendments/bio-stimulants for hydrocarbon-eating microbes attached to support media surfaces in a fixed-bed bioreactor system. CT and ESP were characterized to be rich in essential nutrients including N, P, K, Ca, Mg, Na and C. CT and ESP application at four levels each recorded appreciable boosts in microbial numbers and TPH removal across applied levels. CT application recorded 23.49±2.20 to 29.36±0.79 % boost in TPH removal and 17.03±3.01 to 61.07±2.26 % boost in microbial mass. Boosts in TPH removal assumed the order 3200 ml > 1600 ml > 800 ml > 400 ml. ESP application similarly recorded 27.56±2.62 to 29.57±0.90 % boost in TPH removal and 17.50±1.60 to 24.43±2.05 % boost in microbial mass. Recorded TPH removal boosts assumed the trend 20 g/l >10 g/ l > 2.5 g/l >5 g/l.


Biostimulation; Bioremediation; Hydrocarbon contaminated water; Total Petroleum Hydrocarbon; Hydrocarbon-consuming microbes; Fixed-bed Bioreactor

Full Text:



Satheesh A, Rajakumar S, Ashok, Umadevi A and Akila V. Insight of Various Environmental Derived Polycyclic Aromatic Hydrocarbons (PAHs) and Consequences on Living Organisms from the Soil-A Review. World Journal of Pharmaceutical Research. 2015, 4(2), 369-383

Abioye OP, Agamuthu P and Abdul Aziz AR. Biodegradation of Used Motor Oil in Soil Using Organic Waste Amendments. Biotechnology Research International. 2012, pp. 1-8. doi:10.1155/2012/587041

Das N and Chandran P. Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview. Biotechnology Research International. 2011, pp. 1-13 doi:10.4061/2011/941810

Adelowo OO, Alagbe SO and Ayandele AA. Time-Dependent Stability of Used Engine Oil Degradation by Cultures of Pseudomonas fragi and Achromobacter aerogenes. African Journal of Biotechnology. 2006, 5 (24): 2476-2479

Edewor TI, Adelowo OO and Afolabi TJ. Preliminary Studies into the Biological Activities of a Broad Spectrum Disinfectant Formulated from Used Engine Oil. Poll Res. 2004, 23(4): 581-586

Mandri T and Lin J. Isolation and Characterization of Engine Oil Degrading Indigenous Microorganisms in Kwazulu-Natal, South Africa. African Journal of Biotechnology. 2007, 6(1): pp. 023-027

Varjani SJ and Upasani VN. A New Look on Factors Affecting Microbial Degradation of Petroleum Hydrocarbon Pollutants. International Biodeterioration and Biodegradation, Elsevier, 2017, 120: 71-83, doi:10.1016/j.ibiod.2017.02.006

AbdulRahim AA and Gaber MAI. Biological Treatment of Hydrocarbon Contaminants: Petroleum Hydrocarbon Uptake by Pseudomonas alkanolytica. JKAU: Eng. Sci. 2010, 21(1): 39-53, doi: 10.4197/ Eng. 21-1.3

Lageman R, Clarke RL and Pool W. Electro-reclamation: A Versatile Soil Remediation Solution. Eng Geol. 2005, 77: 191-201

Adams GO, Fufeyin PT, Okoro SE and Ehinomen I. Bioremediation, Biostimulation and Bioaugmentation: A Review. International Journal of Environmental Bioremediation and Biodegradation. 2015, 3(1): 28-39

Joutey NT, Bahafid W, Sayel H and El Ghachtouli N. Biodegradation: Involved Microorganisms and Genetically Engineered Microorganisms: in Biodegradation - Life of Science, Dr. Rolando Chamy (Ed.). InTech, 2013, doi: 10.5772/56194

Hakeem KR, Akhtar J and Sabir M. Soil Science: Agricultural and Environmental Prospectives. Springer International Publishing, 2016, pp. 430, doi: 10.1007/978-3-319-34451-5

Fingerman M and Nagabhushanam R. Bioremediation of Aquatic and Terrestrial Ecosystems. Science Publishers Inc., 2016, pp. 622

Leahy JG and Colwell RR. Microbial Degradation of Hydrocarbons in the Environment. Microbiol. Rev. 1990, 54: 305-315

Fei-Baffoe B, Antwi-Akomeah S, Agyapong EA and Osei-Fosu P. Biodegradation of Used Lubricating Engine Oil Contaminated Water Using Indigenous Hydrocarbon Degrading Microbes in a Fixed Bed Bioreactor System. Journal of the Ghana Science Association. 2012, 14(1): 94-102

Rutala WA, Weber DJ. Healthcare Infection Control Practices Advisory Committee. Guidelines for Disinfection and Sterilization in Healthcare Facilities. CDC Stacks-Public Health Publications, 2008

Fukuzaki S. Mechanisms of Actions of Sodium Hypochlorite in Cleaning and Disinfection Processes. Biocontrol Science, 2006, 11(4): 147-157

Darmady EM, Hughes KEA, Jones JD, Prince D and Tuke W. Sterilization by Dry Heat. Journal of Clinical Pathology. 1961, 14(1): 38-44

Abeka H. Use of Biochar to Enhance Bioremediation of An Oxisol Contaminated with Diesel Oil. 2014, M.phil Theses. Accessed 09-03-2017

Okpokwasili GC and Amanchukwu SC. Petroleum Degradation by Candida species. Environment International. 1988, 14: 243-247

Kastner M, Breuer-Jammali M and Mahro B. Enumeration and Characterization of Soil Microflora from Hydrocarbon contaminated Soil Sites able to Mineralize Polycyclic Aromatic Hydrocarbons (PAH). Applied Microbiol. Biotech. 1995, 41: 267-273

Mills AL, Breuil C and Colwell RR. Enumeration of Petroleum Degrading Marine and Estuarine Microorganisms by the Most Probable Number Method. Canadian Journal of Microbiology. 1978, 24: 552-557

Zajic JE and Supplisson B. Emulsification and Degradation of ‘Bunker C’ Fuel Oil by Microorganisms. Biotechnology and Bioengineering. 1972, 14(3): 331-343

Deepthi, K.P. and Reddy, P.N. Compost Teas: An Organic Source for Crop Disease Management. International Journal of Innovative Biological Research, 2013, 2(1): 51-60

Shih-Ching W, Hsueh-Chuan H, Shih-Kuang H, Ya-Chu C and Wen-Fu H. Synthesis of Hydroxyapatite from Eggshell Powders through Ball Milling and Heat Treatment. Journal of Asian Ceramic Societies. 2016, 4(1): 85-90.

Munter R. Laboratory Factors Affecting the Extractability of Nutrients, pp. 8-10: in Dahnke (edition)-Recommended Chemical Soil Test Procedures for the North central Region, North Dakota Agricultural Experiment Station Bulletin, 1988, No. 399 (revised)

Berg MG and Gardner EH. Methods of Soil Analysis Used in the Soil Testing Laboratory at Oregon State University. Special Report-321, 1978, 1-45

Capar SG Tanner JT, Friedman MH and Boyer KW. Multielement Analysis of Animal Feed, Animal Wastes, and Sewage Sludge. Environmental Science Technology, 1978, 12: 785.

Olsen SR, Cole DV, Watanabe FS and Dean LA. Estimation of Available Phosphorus in Soils by Extraction with Sodium Bicarbonate. USDA Circular No: 939, US Gov. Print Office, 1954, Washington, D.C.

Olsen SR and Dean LA. Phosphorus, Chap. 73 of Methods of Soil Analysis, Part 2. Amer. Soc. Agron., Madison, Wisconsin, 1965, 1035-1048

Allison LE. Organic Carbon: In Methods of Soil Analysis, Black, C.A. (Ed.). American Society of Agronomy, USA, 1965, 1367-1378

Chimezie Dirisu NG. Isolation and Characterization of Hydrocarbon-Utilizing Fungi from Fresh Water Swampy Soil. Microbiology Research International, 2015, 3(2): 33-36, ISSN: 2354-2128.

Chikere CB, Chikere BO and Okpokwasili GC. Bioreactor-based Bioremediation of Hydrocarbon-polluted Niger Delta Marine Sediment, Nigeria. Biotech, 2012, 2(3): 53-66: doi 10.1007/s13205-011-0030-8

Goldman E and Green LH. Practical Handbook of Microbiology. 2nd Edition, CRC Press, 2008, 17-18

Kilduff JE, Komisar S and Nyman M. Microbial Enumeration as a Tool for Bioremediation Monitoring: In Hazardous and Industrial Waste, Proceedings, 32nd Mid-Atlantic Conference. CRC Press, 2000, 63-64

Environmental Research Institute, University of Connecticut. Analysis of Extractable Total Petroleum Hydrocarbons (ETPH) using Methylene Chloride: Gas Chromatograph/Flame Ionization Detection. 1999, 1-10

Suthersan SS. Remediation Engineering: Design Concepts. Lewis Publishers, Boca Raton, FL. 1997

Pant AP, Radovich TJK, Hue NV and Paull RE. Biochemical Properties of Compost Tea Associated with Compost Quality and Effects on Pak Choi Growth. Scientia Horticulturae. 2012, 148: 138-146

Scheuerell SJ and Mahaffee WF. Compost Tea: Principles and Prospects for Plant Disease Control. Compost Sci Util. 2002, 10: 313-338

Salleh AB, Ghazali FM, Abd Rahman RNZ and Basri M. Bioremediation of Petroleum Hydrocarbon Pollution. Indian Journal of Biotechnology. 2003, 2: 411-425

Zhu X, Venosa AD, Suidan MT and Lee K. Guidelines for the Bioremediation of Marine Shorelines and Freshwater Wetlands. Cincinnati: US Environmental Protection Agency, 2001.

Cooney JJ. The Fate of Petroleum Pollutants in Freshwater Ecosystems: in Petroleum Microbiology, R. M. Atlas, Ed. MacMillan, New York, USA, 1984, 399-434

Siulapwa N, Mwambungu A and Mubbunu L. Composition of Commercial Hen Egg Shells to Fresh Water Crocodile Egg Shells. International Journal of Research in Agriculture and Food Sciences. 2014, 2(7): 16-18

Yasothai R and Kavithaa NV. Chemical Characterization of Egg Shell Meal. International Journal of Science, Environment and Technology. 2014, 3(4): 1436-1439

Van Herwijnen R, Joffe B, Ryngaert A, Hausner M, Springael D, Govers HAJ, Wuertz S and Parsons JR. Effect of Bioaugmentation and Supplementary Carbon Sources on Degradation of Polycyclic Aromatic Hydrocarbons by a Soil-derived Culture. FEMS Microbiology Ecology. 2006, 55: 122-135. doi:10.1111/j.1574-6941.2005.00001.x

Bayat Z, Hassanshahian M and Cappello S. Immobilization of Microbes for Bioremediation of Crude Oil Polluted Environments: A Mini Review. The Open Microbiology Journal, 2015, 9: 48-54

Martins SCS, Martins CM, Guedes Fiúza LMC and Santaella ST. Immobilization of Microbial Cells: A Promising Tool for Treatment of Toxic Pollutants in Industrial Wastewater. African Journal of Biotechnology. 2013, 12(28): 4412-4418, doi: 10.5897/AJB12.2677

Górecka E and Jastrzębska M. Immobilization Techniques and Biopolymer Carriers. Biotechnol. Food Sci. 2011, 75: 65-86

Braschler T, Johann R, Heule M, Metref L and Renaud P. Gentle Cell Trapping and Release on a Microfluidic Chip by In-situ Alginate Hydrogel Formation. Lab Chip. 2005, 5: 553-559.

Atlas RM. Biological Degradation of Petroleum Hydrocarbons: An Environmental Perspective. Microbiol. Rev. 1981, 45: 180-209

Alexander M. Biodegradation and Bioremediation. 2nd Edition, Academic Press, San Diego, 1999, 269-298

Atlas RM. Microbiology-Fundamentals and Applications. 2nd Edition, Macmillan Publishing Co., New York, 1988, 352-353

Hambrick GA, DeLaune RD and Patrick Jr, WH. Effect of Estuarine Sediment pH and Oxidation-reduction Potential on Microbial Hydrocarbon Degradation. Applied Environmental Microbiology. 1980, 40: 365-369

Dibble JT and Bartha R. Effect of Environmental Parameters on the Biodegradation of Oil Sludge. Appl. Environ. Microbiol. 1979, 37: 729-739

Verstraete W, Vanloocke R, Deorger R and Verlinde A. Modelling of the Breakdown and the Mobilization of Hydrocarbons in Unsaturated Soil Layers, pp. 99-112: In J.M. Sharpley and A.M. Kaplan (ed.), Proceedings of the 3rd International Biodegradation Symposium, Applied Science Publishers Ltd., London, 1976

Prescott ML, Harley JP and Klan AD. Industrial Microbiology and Biotechnology: In Microbiology. 3rd. Ed., Wim C. Brown Publishers, Chicago. 1996, pp., 923-927

Olajire AA and Essien JP. Aerobic Degradation of Petroleum Components by Microbial Consortia. Journal of Petroleum and Environmental Biotechnology. 2014, 5:195, doi: 10.4172/2157 7463.1000195

Atlas RM and Bartha R. Biodegradation of Petroleum in Seawater at Low Temperatures. Canadian Journal Microbiology. 1972, 18:1851-1855

Walkley A and Black IA. An examination of the Degtjareff Method for Determining Soil Organic Matter and a Proposed Modification of the Chromic Acid Titration Method. Soil Science. 1934, 37: 29-38


  • There are currently no refbacks.

AJEST (ISSN 2689-1964) Copyright © 2012-2021. All rights reserved. Published by Ivy Union Publishing, 3204 Valley Rush Dr, Apex, North Carolina 27502, United States