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Nitrogen, Phosphorus, Potassium Status of Bioaugmented Versus Biostimulated Crude Oil Contaminated Coastal Plain Sand of Southern Nigeria. (1) Etukudoh,

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Presentation on theme: "Nitrogen, Phosphorus, Potassium Status of Bioaugmented Versus Biostimulated Crude Oil Contaminated Coastal Plain Sand of Southern Nigeria. (1) Etukudoh,"— Presentation transcript:

1 Nitrogen, Phosphorus, Potassium Status of Bioaugmented Versus Biostimulated Crude Oil Contaminated Coastal Plain Sand of Southern Nigeria. (1) Etukudoh, Ndarake Emmanuel (2) Ikpe, F. N (3) Akpan, Joyce Fidelis (4) Ayolagha, Gaskin (1,2) Department of Soil Science, Faculty of Agric., Rivers State University of Science and Technology, PMB 5080, Port Harcourt, Rivers State Nigeria. (3 )Department of Soil Science, Faculty of Agriculture. University of Calabar.Cross River State (4 )Department of Soil Science, Faculty of Agriculture, Niger Delta University. Wilberforce Island, Bayelsa State.

2 INTRODUCTION Biochemical Effects of crude oil Spill on the environment notwithstanding our natural ecosystem has been subjected to series of disturbance crude oil exploration and exploitation by man. Overpopulation and industrialization have also contributed in various ways to the deterioration of the environment upon which humanity is completely dependent for life (Odu, 1980). Crude oil contamination on a soil environment is known to affect soil quality via changes in soil pH, air temperature, soluble plant nutrients and disruption of metabolic processes of soil microorganisms (Moller, 1991, Isirimah, 2004) hence, the vigor of plant nutrients mineralization. As early as 1930, Scientists in Moscow noted the decreases in growth rate of local trees and attributed it to wide spread oil pollution and began five-year remediation plan. In a follow up experiment, George Robinson (known as father of bioremediation) experimented the action of soil microorganisms in a crude oil polluted soil in Santa Maria, California and achieved a huge success (Meagher, 2000). Since then bioremediation has been judged the best alternative method to remedy hydrocarbon contaminated soil. Methods of bioremediation include bioaugmentation (seeding of hydrocarbonoclastic organisms into the contaminated soil) and Biostimulation (provision of appropriate microbial food to stimulate indigenous microbial growth).

3 Materials and Methods Pot experiment was conducted in Green-house of the Rivers State University of Science and Technology Teaching and Research farm, Port Harcourt in the Rivers State of Nigeria. It is located on latitude 4 o 50 1 N and longitudes 7 o 01 1 E and on elevation of 18m above mean sea level (FAO,1984). The area experiences two distinct seasons (raining and dry season). The raining season starts from April and lasts till October with a brief period of dryness (August Break). The rainfall is heavy with estimated annual range which may vary from 2000 to 2484mm (FAO, 1984, MANR, 2005). Rainfall pattern is bimodal with peaks in June and September (Ukpong, 1992). Top soil (0-15cm) was excavated from the uncultivated area of the Rivers State University of Science and Technology Teaching and Research farm for the experiment. Forty five (45) kilograms of soil were respectively polluted to 0,2,5 and 10% with Bonny light crude oil of 0.835 relative density(specific gravity) and left undisturbed for seven days to settled, then fifteen (15) kilograms of each of the pollution level were respectively amended with 100kgha- 1 Urea, 10tha -1 poultry manure and left unamended. Three (3) kilograms of each of the treatment options were transferred into five (5) poly bags measuring 30x25cm 2 and seeded with 130 litres of 48 hours nutrient broth of Acinetobacter clavatus, Bacillus subtilis, Pseudomonas aeruginosa and Corynebacterium diphtheriae, left unseeded and replicated three times, then completely randomized and arranged in a Greenhouse. The experiment was kept moistened at field capacity during the study period (Page et al., 1982).

4 Soil Sampling Before treatments, soil samples were analyzed for physical and chemical properties. At week nine (9), soil was sampled for percent total nitrogen, a vailable phosphorus and potassium. Analytical methods for soil Soil pH was determined in water with glass electrodes in 1:2.50 soil water ratio (Udo and Ogunwale, 1978); organic matter was determined by methods of Nelson and Summers (1982): Total nitrogen was determined by methods described by Udo et al., (2009); Calcium (Ca) and Magnesium(Mg) were extracted with molar ammonium acetate; Potassium and Sodium concentrations were determined by flame photometry as described by Udo et al.,(2009); Aluminum plus hydrogen, was extracted with molar Potassium chloride(KCL) and acidity determined by titration(Mclean, 1965); effective cation exchange capacity was determined according to Kamprath (1984). Av.P was determined by methods described by Sparks (1996).

5 Statistical Analysis Analysis of variance according to the completely randomized design of 0.05% probability (LSD) level was used to compare the treatments effects.

6 Results Table 1: Physicochemical Properties of the soil used for the study Parameters MeasuredValues pH (H 2 O): ratio 1:2.505.80 % organic carbon3.91 Percent total N0.12 Available phosphorus (mg/kg)267.00 Ca (Cmol/kg)1.30 Mg (Cmol/kg)0.73 Na(Cmol/kg)0.86 K (Cmol/kg)5.96 ECEC1.00 %BS83.22 % sand858.20 % silt84.20 % clay57.60 Textural classSandy loan

7 Table 2. Chemical Properties of Poultry Manure used as soil amendment ParameterValue pH 6.99 Total N (%) 3.01 Phosphorus (mg/kg) 3893.12 Calcium (cmol/kg) 922.60 Magnesium(cmol/kg) 578.40 Sodium (cmol/kg) 1318.00 Potassium (cmol/kg) 1823.00

8 Table 3: Effect of treatments on soil available phosphorus (Av.P) (ppm) Treatment Week 0 week 9 Crude Oil Level AmendmentsAmendments/Bacteria 0 1 2 0 1 2 0%B­­ o 267 314 388208 430 490 0%B 1 220 227 245189 300 310 0%B 2 218 225 255200 260 330 0%B 3 225 249 260199 284 390 0%B 4 230 250 273210 300 360 2%B o 353 399 434400 480 510 2%B 1 332 363 388360 430 440 2%B 2 309 323 354320 350 400 2%B 3 333 327 384340 370 480 2%B 4 315 336 390332 420 468 5%B o 309 315 387500 580 640 5%B 1 206 233 243435 485 458 5%B 2 210 225 251403 429 463 5%B 3 255 239 268397 440 470 5%B 4 219 206 247434 460 461 ­ 10%B­­­ o 220 242 254240 280 340 10%B 1 208 221 230300 319 396 ­ 10%B 2 183 207 221299 356 378 10%B 3 85 110 227282 393 360 10%B181 199 205309 337 383 LSD (0.05) 15.27 16.32 12.8132.17 24.23 40.30 Key: B 0, B 1, B 2, B 3,­ B 4 = Soil without inculcation, Soil inoculated with A. clavatus, B. Subtilis, P. aeruginsona and C. diphtheriae respectively.:0,1,2, = Soil without amendment, Soil amended with urea and PM respectively

9 Table 4: Effects of treatments on total nitrogen Treatment combinationWeek 0Week 9 Crude Oil LevelAmendmentsAmendment/Bacteria 0 1 2 0%B­­ o 0.12 0.11 0.100.07 0.08 0.09 0%B 1 *0.10 0.12 0.120.04 0.06 0.08 0%B 2 *0.10 0.12 0.110.03 0.05 0.07 0%B 3 *0.10 0.10 0.120.04 0.05 0.06 0%B 4 *0.11 0.11 0.110.04 0.05 0.07 2%B o 0.09 0.08 0.090.12 0.14 0.15 2%B 1 0.09 0.06 0.070.10 0.11 0.12 2%B 2 0.09 0.07 0.080.11 0.08 0.11 2%B 3 0.10 0.08 0.070.09 0.08 0.08 2%B 4 0.10 0.07 0.080.10 0.10 0.11 5%B o 0.08 0.09 0.090.13 0.15 0.16 5%B 1 0.07 0.07 0.080.11 0.13 0.12 5%B 2 0.07 0.06* 0.080.11 0.12 0.12 5%B 3 0.07 0.07* 0.070.11 0.12 0.11 5%B 4 0.07 0.07 0.080.12 0.13 0.13 ­ 10%B­­­ o 0.06 0.08 0.080.08 0.10 0.09 10%B 1 0.06 0.06 0.070.09 0.10 0.10 ­ 10%B 2 0.06 0.06 0.070.09 0.11 0.10 10%B 3 0.06 0.05* 0.060.10 0.12 0.10 10%B 4 0.06 0.06 LSD (0.05) 0.01 0.02 0.020.1 N.S N.S Key: B 0, B 1, B 2, B 3,­ B 4 = Soil without inoculcation, Soil inoculated with A. clavatus, B. Subtilis, P. aeruginsona and C. diphtheriae respectively. :0,1,2, = Soil without amendment, Soil amended with urea and PM respectively. :*= significant, NS= Not significant..

10 Discussions After treatments, lower Av.P values obtained in bacterial seeded soil as compared to that of unseeded soil may be due to immobilization of soil Av.P by the Soil microorganisms utilizing Av.P as bacterial seeding is likely to increase microbial cell in the soil. Also higher Av.P values obtained in the amended soil as compared to the unamended soil suggests lack of nutrient resources to support seeded microbial populations in the unamended soil. Subsequent increase in Av.P at 2% crude oil pollution level in week 9 in comparison to that of 5-10 crude oil pollutant level; therefore mean that at 2% crude oil pollution level effect on soil biochemical processes was minimal at 2%, while at 5-10% some microbial vital functions may have been inhibited in the presence of/and toxicity of high crude oil pollution level as earlier reported by Xu and schurri (1990). In week 9, soil Av.P status improved at all levels of crude oil pollution level. This may be due to subsequent changes in the Soil biochemical processes with time to as degradation proceed. It also means that toxicity of crude oil get reduced with time and that the degradation and mineralization of phosphorus out of hazardous substances also improved with time as earlier reported by Xu and Schurri, (1990). Isrimah et al., (1989) also observed that availability of phosphorus is pH related.

11 Discussion conti. Etukudoh, (2011) observed that the presence of soil amendment (feed stuff) and crude oil significantly stimulated bacterial growth, thus immobilization. Therefore, lower Av.P values in the Urea amended soil as compared to that of PM amended soil in week 9 indicated that the former was more effective in regulating soil reaction (pH) to suite microbial activities. Considering the Study period, this Suggests that PM (organic fertilizer) is better than the Urea (inorganic fertilizer) in making phosphorus more available. Decrease in total N in the soil after bacterial seeding as compared to unseeded soil may be due to increase in bacterial activities. This therefore indicated that bacterial seeding and application of soil amendments were effective for the process of phosphorus mineralization to take place. Subsequent increase in total N at week 9 during the study period may be due to increased in the rate of mineralization and vigor of nitrification as earlier reported by Odu (1980) and Isirimah et al; (1989). Lower total N values recorded at 2% crude oil pollution level as compared to other pollution levels may be due to the attraction and spontaneous increase in microbial population due to the presence of crude oil and soil amendments. This was earlier observed by Ita and Essien (2002), who reported that at a lower level of crude oil pollution level, hydrocarbon utilizers are attracted and their increase stimulated more than in the highly contaminated or in the uncontaminated soil.

12 Discussion conti. Decreased in soil K in the soil with increase in crude oil pollution level was observed with significant differences among treatments option in both week 1 and in week 9. This indicated that crude oil pollution level as low as 2% could affect K dynamic in the Soil. According to Alexander (1979), the quantity of water soluble non- exchangeable K fraction is microbial in origin. Therefore depression of soil organism due to crude oil and presence of soil amendments and bacterial seeding are synonymous with K depletion in the Soil. In week 9, potassium (K) values were higher in the Urea amended soil. It therefore means that PM could better raise K status in the soil than that of Urea and that bacterial seeding will result in the depletion of soil K.

13 Conclusion Effects of bioaugmentation and biostimulation on the availability of phosphorus, nitrogen, and potassium in a crude oil contaminated Coastal Plain Sands of southern Nigerian were evaluated. Bioaugmentation resulted in the decrease of all tested parameters in all treatment options. Soil seeded with C. diptheriae and A. clavatus, P.aeruginosa made more phosphorus available at 0 and 2, 5 and 10% crude oil pollution levels respectively in all treatment options while values of total N and Potassium were higher in the unseeded soil. In general, Av.P and K values were higher in the PM amended soil, total N values were also higher in the 0- 5% crude oil pollution level amended with PM but at 10% crude oil pollution levels values were higher in the urea amended This study suggests that 2-5% crude oil concentrated Coastal Plain Sands is better amended with PM and Seeded with C. diphtheriae while above 5% crude oil contamination should be amended with Urea and Seeded with P. aeruginosa to bring about much desirable NPK status in the soil..

14 References Alexander, M(1977): Introduction to Soil microbiology, 2 nd Edition, New York, John Wiley and Sons pg 23g. Buchanan. R.V and Gibbons N.E eds(1974): Bergrey’s Mauual of Determinative Bacteriology Ed. Willians and Wilkins Co Bathimox Cowan, S.J. (1974): manual for identification of medical Bacterial. University Press Cambridge. Esu, IE. (1999). Fundamentals of pedology. Sterling Hordes publishers Nig.LTD. Etukudoh, N.E (2011): Evaluation of Biostimulated and Bioaugmented crude oil contaminated Coastal Plain Sand of southern Nig. Journal of Agric. And vet. Serves.vol.3 pg 89.96 FAO (1984): Food and Agriculture Organization. Production Year Book.Vol.4 FAO. Etaly pg 122-127 Harrigan, W. F. and McCane (1990). Laboratory Method in Medical Mycology. 8 th Edition. London; Academic Press, Ibia, T.0 and Udo E.J. (2009). Guide to fertilizer use for crops in Akwa Ibom State. Nigeria. Sibon Books ltd. Lagos Isirimah, N.O, Zuofa K and loganathan (1989): Effect of crude oil on maize performance and Soil chemical properties in Humid forest zone of Nigeria Isirimah, N.O (2004): Soils and Environmental pollution Management. Luton publishers luton press ltd. Owerri. Ita,A. Y and Essien, J.P(2002): Petroleum Capabilities and growth profile of Bacterial from Crude oil polluted Coastal Plain Sand and Blackish water, Global journal of pure and applied services, vol.7.No.3, 507-511 Kamprath, EJ. (1984). Crops Response to liming in the Tropics.Soil Acidity and Linning. Journal 2:349- 366. Hetropics Acidity Klute, A (1986). Methods of Soil Analysis physical and mineralogical properties No.9 (1). American J. of Soil Argon monograph. Madison W.I.USA. MANR (2005). Ministry of Agriculture and Natural Resources Rivers State Nigeria. McLean, EO. (1965). “Aluminum” methods of Soil Analysis part 2 Black, C.A. Argon. No. 9:986-994 Meagher, R.B. (2000). Phytoremediation of toxicElemental and Organic pollutants”Current opinion plant Biology 3(2):153-172.

15 References conti. Mills. A. Bremil, l.c. and colwell, RR (1978).Enumeration of petroleum Degrading marine and Estuarine Microorganisms by probable Number Method. Can J. of microbiology. (24) 552-557 Moller, C.J. (1991). Environmental Science, Sustaining the Earth. Wadswofth Publishers Belmon, C.A. Nelson, DW. And Sommers L.W. (1982). Total carbon, organic C and organic Matter in methods of Soil Analysis part 2 chemical and microbiological No.9 zed. Pg 539-552 Nyle, C.P and Ray, R.W (2007). The Native and properties of Soil (Bth ed), Inc. New York pp156. Odu C.T.I (1980). Degradation of crude oil under Tripical condition. In proceedings of an int’l Serminar on the petroleum Ind. And Nig. Environ. 1980. PTI Warri, NIGERIA proceedings of an int’l Serminar on the petroleum Ind. And Nig. Environ. 1980. P.J.I Warri, NIGERIA. Page, A. l., Miller, I. T and Kenney D. R (1982). Methods of Soil chemical Analysis part 2 chemical and microbiological properties. Org. No. 9,2 nd Edition sparks, D.L. (1996). Methods of Soil Analysis part 3 chemical properties SSSA. Madison W.I. Soil survey staff (1999). Soil Taxonomy. A basic system of Soil classification for making and interpreting Soil surveys. USDA. Agric. Hand book No. 436. 2 nd Ed. Us. Govt. priority office. Washington DC. Pp 868 Sparks, D. L. (1996). Method of Soil Analysis, Part 3, Chemical Propertieiss. SSA Madison, W. A. Udo, E.J. and ogunwale, JA. (1978): laboratory manual for the Analysis of Soil, plants and water samples. Pp 2 - 12, unpublished monograph. Dept. of Agronomy University of Ibadan Nigeria. Udo, E. J.; Ibia, T. O. (2009).Manual of Soil Plant and Water Analyses. Sibon Books Limited. Flat 15, Block 6, Fourth Avenue, Festac, Lagos. Ukpong, I E. (1992). The structure and Soil relations of Avicennia mangrove Swamps in South Eastern Nigeria. Tropical ecology (33: 1-16) Xu. H.H and Schurri, KM (1990). Genotoxicity of 22 pesticides in microtitration. S.O.S Chromotest Toxicity Assessment 1-4

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