Comparison of the Biodegradation of n-alkanes and Readily Biodegradable Substrates Using Open Mixed Culture under Aerobic, Anoxic and Anaerobic Conditions

Chukwuemeka Uzukwu^1, and Davide Dionisi¹

¹Materials and Chemical Engineering Group, School of Engineering, University of Aberdeen, Aberdeen, AB24 2UE, UK

Cite this paper:
Chukwuemeka Uzukwu and Davide Dionisi. Comparison of the Biodegradation of n-alkanes and Readily Biodegradable Substrates Using Open Mixed Culture under Aerobic, Anoxic and Anaerobic Conditions. International Journal of Environmental Bioremediation & Biodegradation. 2017; 5(2):65-76. doi: 10.12691/ijebb-5-2-5

Abstract

This study has investigated the biodegradation of n-alkanes using open mixed cultures in batch tests. Biodegradation of n-C12, C14, C16, C18, C20 was investigated using a respirometric method and compared with the biodegradation of the readily biodegradable substrates glucose, acetic acid and ethanol. Experiments were performed in small-scale bioreactors under various conditions, i.e. aerobic, anoxic with nitrate and completely anaerobic conditions, using two different sources of open mixed microbial cultures. Under aerobic conditions all the readily biodegradable substrates and hydrocarbons were removed, although the acclimation time was longer for the hydrocarbons (3-5 days) than for the readily biodegradable substrates (1-2 days). No significant effect on the hydrocarbon concentration on biodegradation was observed in the concentration range 0.5-5 g/l. Under anoxic conditions, both the readily biodegradable substrates and the hydrocarbons were removed using nitrate as electron acceptor. However, the acclimation time for the hydrocarbons under anoxic conditions (20-25 days) was much longer than under aerobic conditions. Under both aerobic and anoxic conditions, once acclimation with the hydrocarbons was completed, the microorganisms were immediately able to remove a second spike of the substrate without acclimation. Under anaerobic conditions, no activity was observed with the hydrocarbons over a period of 150 days, while the mixed culture was able to remove glucose and convert it to volatile fatty acids. Under aerobic conditions, the dissolved oxygen consumption data was mathematically modelled using Monod kinetics to obtain biokinetic parameters. Good fittings between the model and the experimental data was obtained and the biodegradation of hydrocarbons was characterised by higher values of the parameter K_S compared to the readily biodegradable substrates. The results of this study showed that the open mixed microbial cultures contained diverse microorganisms capable of utilizing both liquid and solid n-alkanes under aerobic and anoxic conditions.

Keywords:
biodegradation n-alkanes mixed culture aerobic anoxic anaerobic respirometry

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References:

[1]	Brassington, K.J., Hough, R.L., Paton, G.I., Semple, K.T., Risdon, G.C., Crossley, J., Hay, I., Askari, K. and Pollard, S.J., (2007). Weathered hydrocarbon wastes: a risk management primer, Critical Reviews in Environmental Science and Technology, 37 (3), pp. 199-232.
[2]	MacLeod, C.J., Morriss, A. and Semple, K.T., (2001). The role of microorganisms in ecological risk assessment of hydrophobic organic contaminants in soils.
[3]	Harayama, S., Kasai, Y. and Hara, A., (2004). Microbial communities in oil-contaminated seawater, Current Opinion in Biotechnology, 15 (3), pp. 205-214.
[4]	Gersberg, R.M., Dawsey, W.J. and Bradley, M.D., (1993). Nitrate enhancement of in‐situ bioremediation of monoaromatic compounds in groundwater, Remediation Journal, 3 (2), pp. 233-245.
[5]	Venosa, A.D., Campo, P. and Suidan, M.T., (2010). Biodegradability of lingering crude oil 19 years after the Exxon Valdez oil spill, Environmental Science & Technology, 44 (19), pp.7613-7621.
[6]	Nicol, J., Wise, W.R., Molz, F.J. and Benefield, L.D., (1994). Modeling biodegradation of residual petroleum in a saturated porous column, Water Resources Research, 30 (12), pp. 3313-3325.
[7]	Sarioglu, M.S. and Copty, N.K., (2008). Modeling the enhanced bioremediation of organic contaminants in pyrite-containing aquifers, Transport in Porous Media, 75 (2), pp. 203-221.
[8]	Monod, J., (1949). The growth of bacterial cultures, Annual Reviews in Microbiology, 3 (1), pp. 371-394.
[9]	Grady, C., (1985). Biodegradation: its measurement and microbiological basis, Biotechnology and Bioengineering, 27 (5), pp. 660-674.
[10]	Dang, J.S., Harvey, D.M., Jobbagy, A. and Grady Jr, C.L., (1989). Evaluation of biodegradation kinetics with respirometric data, Research Journal of the Water Pollution Control Federation, pp. 1711-1721.
[11]	Stieber, M., Werner, P. and Frimmel, F., (1994). Investigations on the microbial degradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soils, Bioreclamation of Chlorinated and Polycyclic Hydrocarbon Compounds Ed.Hinchee, RE and Leeson, A, pp. 164-171.
[12]	Bartram, A.K., Jiang, X., Lynch, M.D., Masella, A.P., Nicol, G.W., Dushoff, J. and Neufeld, J.D., (2014). Exploring links between pH and bacterial community composition in soils from the Craibstone Experimental Farm, FEMS Microbiology Ecology, 87 (2), pp. 403-415.
[13]	Benedek, A. and Heideger, W.J., (1971). Effect of additives on mass transfer in turbine aeration, Biotechnology and Bioengineering, 13 (5), pp.663-684.
[14]	Sheppard, J.D., (1989). Feedback Control and the Continuous Phasing of Microbial Cultures.
[15]	Poster, D.L., Schantz, M.M., Sander, L.C. and Wise, S.A., (2006). Analysis of polycyclic aromatic hydrocarbons (PAHs) in environmental samples: a critical review of gas chromatographic (GC) methods, Analytical and Bioanalytical Chemistry, 386 (4), pp. 859-881.
[16]	Ying, L., Hongwei, J., Huizhen, X. and Li, L., (2006). A new spectrophotometric method for measuring COD of seawater, Journal of Ocean University of China, 5 (2), pp. 137-140.
[17]	Marino, F., Karp, J. and Cooper, D., (1998). Biomass measurements in hydrocarbon fermentations, Biotechnology Techniques, 12 (5), pp. 385-388.
[18]	Dionisi, D., (2017). Biological Wastewater Treatment Processes: Mass and Heat Balances. CRC Press.
[19]	Kindred, J.S. and Celia, M.A., (1989). Contaminant transport and biodegradation: 2. Conceptual model and test simulations, Water Resources Research, 25 (6), pp. 1149-1159.
[20]	Gödeke, S., Vogt, C. and Schirmer, M., (2008). Estimation of kinetic Monod parameters for anaerobic degradation of benzene in groundwater, Environmental Geology, 55 (2), pp. 423-431.
[21]	Mohamed, M. and Hatfield, K., (2011). Dimensionless parameters to summarize the influence of microbial growth and inhibition on the bioremediation of groundwater contaminants, Biodegradation, 22 (5), pp. 877-896.
[22]	Metcalf, E., (1991). Wastewater Engineering: Treatment, Disposal, and Reuse, 3
[23]	Bailey, J. and Ollis, D., (1986). Applied enzyme catalysis, Biochemical Engineering Fundamentals, 2 pp. 157-227.
[24]	Molz, F.J., Widdowson, M. and Benefield, L., (1986). Simulation of microbial growth dynamics coupled to nutrient and oxygen transport in porous media, Water Resources Research, 22 (8), pp. 1207-1216.
[25]	Schirmer, M., Molson, J.W., Frind, E.O. and Barker, J.F., (2000). Biodegradation modelling of a dissolved gasoline plume applying independent laboratory and field parameters, Journal of Contaminant Hydrology, 46 (3), pp. 339-374.
[26]	Hao, O.J., Richard, M.G., Jenkins, D. and Blanch, H.W., (1983). The half‐saturation coefficient for dissolved oxygen: A dynamic method for its determination and its effect on dual species competition, Biotechnology and Bioengineering, 25 (2), pp. 403-416.
[27]	Bruce, E.R. and Perry, L.M., (2001). Environmental biotechnology: principles and applications, New York: McGrawHill, 400.
[28]	Fritsche, W. and Hofrichter, M., (2008). Aerobic degradation by microorganisms, Biotechnology Set, Second Edition, pp. 144-167.
[29]	Olson, J.J., Mills, G.L., Herbert, B.E. and Morris, P.J., (1999). Biodegradation rates of separated diesel components, Environmental Toxicology and Chemistry, 18 (11), pp. 2448-2453.
[30]	Lal, B. and Khanna, S., (1996). Degradation of crude oil by Acinetobacter calcoaceticus and Alcaligenes odorans, Journal of Applied Bacteriology, 81 (4), pp. 355-362.
[31]	Setti, L., Pifferi, P.G. and Lanzarini, G., (1995). Surface tension as a limiting factor for aerobic n‐alkane biodegradation, Journal of Chemical Technology and Biotechnology, 64 (1), pp. 41-48.
[32]	Thomas, J.M., Yordy, J.R., Amador, J.A. and Alexander, M., (1986). Rates of dissolution and biodegradation of water-insoluble organic compounds, Applied and Environmental Microbiology, 52 (2), pp. 290-296.
[33]	Ladygina, N., Dedyukhina, E. and Vainshtein, M., (2006). A review on microbial synthesis of hydrocarbons, Process Biochemistry, 41 (5), pp. 1001-1014.
[34]	Zilber, I.K., Rosenberg, E. and Gutnick, D., (1980). Incorporation of P and Growth of Pseudomonad UP-2 on n-Tetracosane, Applied and Environmental Microbiology, 40 (6), pp. 1086-1093.
[35]	Miller, T.L. and Johnson, M.J., (1966). Utilization of normal alkanes by yeasts, Biotechnology and Bioengineering, 8 (4), pp. 549-565.
[36]	Bai, G., Brusseau, M.L. and Miller, R.M., (1997). Biosurfactant-enhanced removal of residual hydrocarbon from soil, Journal of Contaminant Hydrology, 25 (1), pp. 157-170.
[37]	Beal, R. and Betts, W., (2000). Role of rhamnolipid biosurfactants in the uptake and mineralization of hexadecane in Pseudomonas aeruginosa, Journal of Applied Microbiology, 89 (1), pp. 158-168.
[38]	Churchill, P.F. and Churchill, S.A., (1997). Surfactant-enhanced biodegradation of solid alkanes, Journal of Environmental Science & Health Part A, 32 (1), pp. 293-306.
[39]	Thibault, G. and Elliott, N., (1979). Accelerating the biological clean up of hazardous materials spills, Proceedings Oil and Hazards Materials.Spills: Prevention-Control-Cleanup-Recovery-Disposal.
[40]	Peng, Y., Yong, M. and Wang, S., (2007). Denitrification potential enhancement by addition of external carbon sources in a pre-denitrification process, Journal of Environmental Sciences, 19 (3), pp. 284-289.
[41]	O’Reilly, C. and Colleran, E., (2005). Toxicity of nitrite toward mesophilic and thermophilic sulphate-reducing, methanogenic and syntrophic populations in anaerobic sludge, Journal of Industrial Microbiology and Biotechnology, 32 (2), pp. 46-52.
[42]	So, C.M. and Young, L.Y., (2001). Anaerobic biodegradation of alkanes by enriched consortia under four different reducing conditions, Environmental Toxicology and Chemistry, 20 (3), pp. 473-478.
[43]	Callaghan, A.V., Tierney, M., Phelps, C.D. and Young, L.Y., (2009). Anaerobic biodegradation of n-hexadecane by a nitrate-reducing consortium, Applied and Environmental Microbiology, 75 (5), pp. 1339-1344.
[44]	Grady Jr, C.L., Daigger, G.T., Love, N.G. and Filipe, C.D., (2011). Biological Wastewater Treatment. CRC press.
[45]	Jones, D., Head, I., Gray, N., Adams, J., Rowan, A., Aitken, C., Bennett, B., Huang, H., Brown, A. and Bowler, B., (2008). Crude-oil biodegradation via methanogenesis in subsurface petroleum reservoirs, Nature, 451 (7175), pp. 176-180.
[46]	Wang, L., Mbadinga, S., Li, H., Liu, J., Yang, S. and Mu, B., (2010). Anaerobic degradation of petroleum hydrocarbons and enlightenment of the prospects for microbial bio-gasification of residual oil, Microbiology, 37, pp. 96-102.
[47]	Aichinger, G., Grady Jr, C.L. and Tabak, H.H., (1992). Application of respirometric biodegradability testing protocol to slightly soluble organic compounds, Water Environment Research, pp. 890-900.
[48]	Van Hamme, J. and Ward, O., (2001). Volatile hydrocarbon biodegradation by a mixed-bacterial culture during growth on crude oil, Journal of Industrial Microbiology and Biotechnology, 26 (6), pp. 356-362.
[49]	Plohl, K., Leskovsek, H. and Bricelj, M., (2002). Biological degradation of motor oil in water, Acta Chimica Slovenica, 49 (2), pp. 279-290.
[50]	Walker, J.D. and Colwell, R.R., (1974). Microbial petroleum degradation: use of mixed hydrocarbon substrates, Applied Microbiology, 27 (6), pp. 1053-1060.
[51]	Grady, J.(.L. and Lim, H.C., (1980). Biological Wastewater Treatment: Theory and Applications. Marcel Dekker, Incorporated.
[52]	Barford, J.P. and Hall, R., (1981). A mathematical model for the aerobic growth of Saccharomyces cerevisiae with a saturated respiratory capacity, Biotechnology and Bioengineering, 23 (8), pp. 1735-1762.
[53]	Bijkerk, A. and Hall, R., (1977). A mechanistic model of the aerobic growth of Saccharomyces cerevisiae, Biotechnology and Bioengineering, 19 (2), pp. 267-296.
[54]	Peringer, P., Blachere, H., Corrieu, G. and Lane, A., (1974). A generalized mathematical model for the growth kinetics of Saccharomyces cerevisiae with experimental determination of parameters, Biotechnology and Bioengineering, 16 (4), pp. 431-454.
[55]	Henze, M., Gujer, W., Mino, T. and Van Loosdrecht, M., (2000). Activated Sludge Models ASM1, ASM2, ASM2d and ASM3. IWA publishing.
[56]	Geng, X., Boufadel, M.C. and Wrenn, B., (2013). Mathematical modeling of the biodegradation of residual hydrocarbon in a variably-saturated sand column, Biodegradation, 24 (2), pp. 153-163.
[57]	Choi, D.H., Hori, K., Tanji, Y. and Unno, H., (1999). Microbial degradation kinetics of solid alkane dissolved in nondegradable oil phase, Biochemical Engineering Journal, 3 (1), pp. 71-78.
[58]	Mulder, H., Breure, A. and Rulkens, W., (2001). Prediction of complete bioremediation periods for PAH soil pollutants in different physical states by mechanistic models, Chemosphere, 43 (8), pp. 1085-1094.