Biodiversity and Dynamics (Rate of Change) of Bacterial Communities Involved in the Biodegradation of Petroleum Refinery Sludge in Contaminated Soils

Main Article Content

Tudararo-Aherobo Laurelta
Atuayan Ernest
Adetutu Eric
Ball Andrew

Abstract

Aim: The study assessed the bio treatability of the petroleum refinery sludge in contaminated soils by indigenous bacterial communities and the effects of the sludge contamination and bio stimulants on the biodiversity and dynamics (rate of change) of the bacterial communities involved in the biodegradation of the sludge, using the molecular biology technique, Denaturing Gradient Gel Electrophoresis (DGGE).

Study Design: The randomnized block design was used for the study.

Place and duration of the Study: The research was conducted in the biology laboratory of Flinders University, Adelaide, South Australia.

Methodology: The percentage of total petroleum hydrocarbons (TPH) degraded and the bacterial load in the test microcosms was assessed tri-weekly for 12 weeks. The percentage TPH was assessed using Gas chromatography, while the bacterial count was determined as gene copies using the culture independent molecular tool, quantitative real-time PCR (qRt-PCR) analysis. The effects of the experimental treatments on the biodiversity and dynamics (rate of change) of the bacterial communities involved in the biodegradation of the sludge in the soils was determined by the culture-independent molecular biology technique, DGGE. Moving Windows Analysis (MWA) and Shannon Weaver diversity index were used to determine the dynamics (rate of change) and biodiversity of the bacterial communities respectively.

Results: Results obtained for the Moving Window Analysis (MWA) which is used to determine the dynamics (Dy), or rate of change of the bacterial communities, showed that, the 1% and 5% sludge contaminated soils biostimulated with compost, recorded the highest Dy of 86.0 ± 1.90% and 87.0 ± 2.20% respectively.NPK biostimilated soil microcosms however recorded a lower Dy of 33.75± 3.20 and 32.50 ± 4.68% for 1% and 5% sludge contamination respectively. The biodiversity of the bacterial communities expressed as Shannon -Weaver index (H1), recorded the highest value of 2.76 ±0.02 for the compost biostimulated microcosm in the 1% sludge treatment, while for the 5% sludge contamination, the treatment with NPK and surfactant enhanced the bacterial biodiversity most with a value of 2.76 ±0.07%. In the test soils with 1% sludge contamination, bio stimulation with NPK gave the highest % TPH degradation (78.25%) while the treatment with NPK and Triton-X 100 had the highest TPH degradation (46.55%) for the 5% sludge contaminated soils. There was insignificant difference in the % sludge degradation between the control and other treatments at P > 0.05 and F = 4.07 for the 1% sludge treated soils, while for the soils treated with 5% sludge there was significant difference between the control and other treatments at P < 0.05 and F= 4.07.

Conclusion: Bacteria species identified in the sludge by molecular biology techniques included; Pseudomonas sp. ITRI77, Uncultured Thauera sp., Uncultured Pseudomonas sp., Flavobacterium sp., Bacillaceae bacterium, Uncultured soil bacterium, Clostridium sp., most of which are Gram negative. Biostimulation with compost enhanced a higher biodiversity (H i) and dynamics (Dy) of the bacterial communities involved in the biodegradation of the sludge. Though the NPK treated soils enhanced the biodegradation of the sludge most, degradation started declining by the 9th week while that of compost continued to rise steadily till the 12th week. Results obtained indicate that compost is as good as NPK in the biodegradation of petroleum sludge especially at 1% sludge contamination, since there was no statistical difference between the % TPH degraded and the use of compost is environmentally friendly and economically sustainable.

Keywords:
Petroleum refinery sludge, biostimulation, bacterial biodiversity, bacterial dynamics (Rate of change).

Article Details

How to Cite
Laurelta, T.-A., Ernest, A., Eric, A., & Andrew, B. (2020). Biodiversity and Dynamics (Rate of Change) of Bacterial Communities Involved in the Biodegradation of Petroleum Refinery Sludge in Contaminated Soils. Journal of Advances in Biology & Biotechnology, 23(4), 23-38. https://doi.org/10.9734/jabb/2020/v23i430150
Section
Original Research Article

References

Ururahy AFP, Marins MDM, Vital RL, Therezinha I, Pereira N. Jr. Effect of aeration on biodegradation of petroleum wastes. Reviews in. Microbiology. 1998; 29: 4.

Hann WJ, Loehr RC. Biological treatment of petroleum oily sludges. SPE –Society of Petroleum Engineers –Periwan Basin oil and Gas Recovering conference, Texas. 1992;519-530.

Louvisse AM, Freire T, Teixeira NO. Metodologia para caracteizacao Borras de petroleoI Mesa Rendonda Sobre Quimica Analitica Ambiental, Curitiba; 1994.

Prince M, Sambasivam Y. Bioremediation of petroleum wastes from the refining of lubricant oils. Environmental Progress. 1993; 12:5-11.

Prospt TL, lochmiller RI, Qualls CW, Jr. Mcbee K. In situ (mesocosm) assessment of immunotoxicity risks to small mammals inhabiting petrochemical waste sites. Chemosphere. 1999;38:1049-1067.

Atlas RM, Bartha R. Hydrocarbon biodegradation and oil spill bioremediation. K.C. Marshall (Ed). Advances in Microbial Ecology. New York: Plenum Press. 1992; 12:287–338.

Mishra S, Jyot J, Kuhad RC, Lal B. Evaluation of inoculum addition to stimulate in situ bioremediation of oily-sludge-contaminated soil’, Applied and Environmental Microbiology. 2001;67: 1675–1681.

Englert CJ, Kenzie EJ, Dragun J. Bioremediation of petroleum products in soil. Principles & Practice of. Petroleum Contamination, in Soils, API. 1993;40:111-129.

Hazardous Waste Management System. Environmental Protection Agency. 1997; 62(29):12.

Oolman T, Baker RR, Renfro NL, Marshall GE. Refinery uses bioslurry process to treat RCRA wastes. Hydrocarbon Processing. 1996;71-76.

Department of petroleum resources. Environmental Guidelines and Standards for the Petroleum Industry in Nigeria (EGASPIN). 2002;314.

Norris RD. Handbook of bioremediation. CRC, Boca Raton; 1994.

Caplan JA. The world-wide bioremediation industry: prospects for profit. Trends Biotechnology. 1993;11:320-323.

Bartha R. Biotechnology of petroleum pollutant biodegradation. Microbiology and Ecology. 1986;12:155–172.

Diplock EE, Mardlin DP, Killham KS, Paton GI. Predicting bioremediation of hydro-carbons: Laboratory to field scale. Environmental Pollution. 2009;157:1831-1840.

Iwamoto T, Nasu M. Current bioremediation practice and perspective. Journal of Bioscience and Bioengineering. 2001;92:1-8.

Kozdroj J, Elsas J. Structural diversity of microorganisms in chemically perturbed soil assessed by molecular and cytochemical approaches. Journal of Microbiological Methods. 2001;43:197-212.

Ranjard L, Poly F, Nazaret S. Monitoring complex bacterial communities using culture-independent molecular techniques: application to soil environment. Research in Microbiology. 2000;167-177.

Tudararo-Aherobo Laurelta, Atuayan Ernest. Toxicological effects of petroleum refinery sludge on the terrestrial environment using bacteria and earthworm as bioindicators. Journal of Advances in Microbiology. 2020; 20(4):30-40.

Mills AL, Breuil C, Colwell RR. Enumeration of petroleum-degrading marine and estuarine microorganisms by the most probable number method. Canadian Journal of Microbiology. 1978; 24(5):522-7.

Walworth J, Pond A, Snape I, Rayner J, Ferguson S, Harvey P. Nitrogen requirements for maximizing petroleum bioremediation in a sub-Antarctic soil. Cold Regions Science and Technology. 2007; 48:84-91.

Namkoong WE, Hwang JP, Choi J. Bioremediation of diesel-contaminated soil with composting. Environmental. Pollution. 2002;119:23.

Zhongun M. Bioremediation of petroleum contaminated soils using indigeneous cultures. A thesis submitted to school of graduate studies, in partial fulfilment of the requirements in master’s of engineering, Faculty of engineering and applied science, Memorial University of Newfoundland, Canada; 1998.

International Organization for Standardi-zation. ISO 16703:2004. Soil quality-determination of content of hydrocarbon in the range C10-C40 by gas chromatography. Geneva, Switzerland: ISO; 2004.

Muyzer G, Smalla K. Application of denaturing gradiient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie van Leewenhoek. 1998; 73:127-141.

Rebrikov DV, Trofimov, Yu D. Real-time PCR: A review of approaches to data analysis. Applied Biochemistry and Microbiology. 2006;42(5):455– 463.

Muyzer G, Waal E, Utterlinden A. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis pf polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology. 1993;695-700.

Sheffield V, Cox D, Lerman L, Myers R. Attachment of a 40-base-pair G+C rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proceedings of the National Academy of Sciences. 1989;86: 232-236.

Anderson IC, Campbell CD, Prosser JI. Diversity of fungi in organic soils under a moorland-Scots pine (Pinus sylvestris L.) gradient. Environmental Microbiology. 2003a;5:1121-1132.

Aroca A, Raposo R. PCR-based strategy to detect and identify species of Phaeoacremonium causing grapevine diseases. Applied Environmental Micro-biology. 2007;73:2911-2918.

Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS. 'Touchdown' PCR to circumvent spurious priming during gene amplification. Nucleic Acids Research. 1991;19.

Teske A, Wawer C, Muyzer G, Ramsing NB. Distribution of sulfate-reducing bacteria in a stratified fjord (Mariagerfjord, Denmark) as evaluated by most-probable-number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments. Appl. Environ Microbiol. 1996;62:1405–141.

Adetutu EM. The fate of azoxystrobin in soils and its effects on soil bacterial and fungal communities, Department of Biological Sciences, University of Essex; 2005.

Girvan MS, Bullimore J, Pretty JN, Osborn AM, Ball AS. Oil type is the primary determinant of the composition of the total and active bacterial communities in arable soils. Applied Environmental Microbiology. 2003;69:1800-1809.

Marzorati M, Wittebolle L, cBoon N, Daffonchio D, Verstraete W. How to get more out of molecular fingerprints: practical tools for microbial ecology. Environmental Microbiology. 2008;10(6): 1571-81. Mills M, McDonald T, Bonner J, Simon M, Autenrieth R. Method for quantifying the fate of petroleum in the environment. Chemosphere. 1999;39: 2563-2582.

Nduka JK, Umeh LN, Okerulu IO, Umedum LN, Okoye HN. Utilization of different microbes in bioremediation of hydrocarbon contaminated soils stimulated with inorganic and organic fertilizers. Journal of Petroleum & Environmental Biotechnology. 2012;3:2.

DOI: 10.4172/2157-7463.100016

Texas Research Institute, Inc. Enhancing the microbial degradation of underground gasoline by increasing available oxygen. Report to the American Petroleum Institute. Washington, D.C; 1982a.

KiIlham K. Soil ecology. Cambridge University Press. U.K; 1994.

Chaudhry GR. Biological degradation and bioremediation of toxic chemicals. Dioscorides Press, Portland, Oregon; 1994.

McBride MB. Toxic metals in sewage sludge-amended soils: Has promotion of beneficial use discounted the risks? Advance Environmental Resources. 2003; 8:5-19.

Efroymson RA, Alexander M. Biodegradation by an arthrobacter species of hydrocarbons partitioned into an organic solvent. Applied and Environmental Microbiology. 1991;57:1441-1444.

Mulligan CN, Yond RN, Gibbs BF. Surfactant-enhanced remediation of contaminated soil: A review. Environmental Geology. 2001;60:371-380.

Lai CC, Huang YC, Wei YH, Chang JS. Biosurfactant-enhanced removal of total petroleum hydrocarbons from contaminated soil. Journal of. Hazardous. Materials. 2009;167:609–614.

Nguyen TT, Youssef NH, McInerney MJ, Sabatini DA. Rhamnolipid biosurfactant mixtures for environmental remediation. Water Resources. 2008;42:1735-1743.

Wittebolle L, Boon N, Vanparys B, Heylen K, De Vos P, Verstraete W. Failure of the ammonia oxidation process in two pharmaceutical wastewater treatment plants is linked to shifts in the bacterial communities. Journal of Applied Microbiology. 2005;99:997–1006.

Schulz S, Giebler J, Chatzinotas A, Wick LY, Fetzer I, Welzl G. Plant litter and soil type drive abundance, activity and community structure of alkB harbouring microbes in different soil compartments. ISME J. 2012;6:1763–1774.

Van Hamme JD, Singh A, Ward OP. Recent advances in petroleum micro-biology. Microbiology and Molecular Biology Reviews. 2003;67(4):503–549.

Okpokwasili GC, Oton NS. Comparative applications of bioreactor and shake flask systems in the laboratory treatment of oily sludge. International. Journal of Environment and Waste Management. 2006;1:1.