Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol.

XXXIX 2009

MBUNTTIREA BIOREMEDIERII SOLURILOR POLUATE CU IEI PRIN UTILIZAREA UNUI PRODUS NATURAL BIODEGRADABIL ENHANCED BIOREMEDIATION OF SOILS POLLUTED WITH CRUDE OIL USING A NATURAL BIODEGRADABLE PRODUCT
MARIANA MARINESCU, M. DUMITRU, ANCA LCTUU, GEORGIANA PLOPEANU

Key words: bioremediation, crude oil, polluted soils, natural biodegradable product. Cuvinte cheie: bioremediere, titei, soluri poluate, produs natural biodegradabil.

REZUMAT Poluarea accidentala cu titei a devenit in zilele noastre un fenomen obisnuit si poate cauza catastrofe ecologice si sociale. Bioremedierea solurilor poluate cu titei este limitata de activitatea microorganismelor degradatoare de hidrocarburi. Principalul scop al acestui studiu este acela de a creste bioremedierea solurilor poluate cu titei prin adaugarea unui produs biodegradabil natural si a inoculului bacterian. Biodegradarea a fost cuatificata prin analize de total hidrocarburi din petrol (THP). Lucrarea prezinta date obtinute in urma procesului de biodegradare a unui sol poluat artificial cu 5% si 10% petrol, tratat cu un produs natural biodegradabil si inocul bacterian. Procesul de biodegradare are loc in timp pentru a reutiliza solul in scopuri agricole. ABSTRACT Crude oil pollution accidents are nowadays become a common phenomenon and have caused ecological and social catastrophes. Crude oil bioremediation of soils is limited by the bacteria activity in degrading the spills hydrocarbons. The aim of this study is to enhance the bioremediation of soils polluted with crude oil by adding the natural biodegradable product and bacterial inoculum. Biodegradation was quantified by total petroleum hydrocarbons (TPH) analyses. The paper presents data obtained in biodegradation process of an artificial polluted soil with 5% and 10% crude oil, treated with a natural biodegradable product and bacterial inoculum. Biodegradation process takes time to reuse the soil in agricultural scopes. INTRODUCTION The development of petroleum industry, and also extractive and processing one, including crude oil transport could lead to unpredictable secondary phenomena with effects more or less harmful for environment and human life. One of this phenomenon is soil pollution with crude oil, petroleum products with or without salty water, petroleum residual products, resulted from crud oil exploitation and extraction (Ru, 1986). In accordance with this phenomenon, starting with 1991, the Institute for Soil Science, Agrochemistry and Environmental Protection ICPA, Bucharest initiated a study in all country. This study had like propose to elucidate the pollution nature, its extension and its impact on soil and microorganisms existing in soil (Toti, 1999). In the study were identified and isolated the main heterotrophic bacteria and microfungi biodegraders of crude oil and assigned some conditions to stimulate their proliferation. Were selected some agro-pedo-improvements practices and biological management to contribute and action on pollutant disappearance from soil (Voiculescu, 2002). 239

Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009

All these research lead to the elaboration of a bioremediation method in-situ of soil polluted with crude oil by using selected microorganisms, with a high efficiency in biodegradation, low costs, without affecting soil characteristics, fertility (Toti, 2003). Petroleum fuel and crude oil products represent the one of the most common environmental contaminant. Common sources of these products are motor fuel station underground storage tanks, home and commercial heating oil storage tanks, fuel distribution centers, refineries, crude oil production sites and accidental spills (Bartha, 1986). Based upon the available information on the chemistry and toxicology of petroleum hydrocarbons, it is possible that petroleum hydrocarbon fuels and oils are mainly composed of aliphatic and aromatic hydrocarbon compounds and petroleum aromatic hydrocarbons generally appear to be more toxic than petroleum aliphatic compounds. Petroleum hydrocarbon products are mixtures of over 250 hydrocarbon compounds. The various product mixtures produced by the manufacturers are based upon physical and performance-based criteria and not specific formulas. As a result, the product compositions can vary depending upon, in part, the crude oil refined to produce the product, the type of product, the season of the year, and any performance additives (Alexander, 1994). Because high concentrations (>1%) of petroleum hydrocarbons in soil have demonstrated phytotoxic properties, as well as the uncertainty about the fate and transport of high concentrations of petroleum in soils and upon human health, the specific fuel nondefault closure levels have maximum limits applied. It is generally recognized that the toxicity, (human and environmental), of petroleum products increases as the molecular weights of the compounds decrease (Pepper, 1996). MATERIALS AND METHODS The main objective of the present study was to enhance the bioremediation method of the soils polluted with crude oil by using the natural biodegradable product ECOSOL. It has been achived an experiment in Green House. The soil was artificial polluted with 5% and 10% crude oil, then was applied the treatment consisting in ECOSOL and bacterial inoculum. The experiment has 11 experimental variants with soil polluted 5% and 10% crude oil, treated with 50g, 100g and 200g ECOSOL/20 kg polluted soil, inoculated and uninoculated with bacteria selected according to the following experimental scheme: V1, control (unpolluted soil); V2, polluted soil with 5% crude oil; V3, polluted soil with 10% crude oil; V4, polluted soil with 5% crude oil + 50 g ECOSOL; V5, polluted soil with 5% crude oil + 50 g ECOSOL + bacterial inoculum; V6, polluted soil with 5% crude oil + 100 g ECOSOL; V7, polluted soil with 5% crude oil + 100 g ECOSOL + bacterial inoculum; V8, polluted soil with 10% crude oil + 100 g ECOSOL; V9, polluted soil with 10% crude oil + 100 g ECOSOL + bacterial inoculum; V10, polluted soil with 10% crude oil + 200 g ECOSOL; V11, polluted soil with 10% crude oil + 200 g ECOSOL + bacterial inoculum. The soil used in experiment is a cambic chernozem sampled from Teleorman area. Chemical characteristics of the soil used in the experiment are presented in table 1.

240

Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009

Table 1 Soil type Calcic chernozem Chemical characteristics of the soil pH Organic Carbon (%) Total Nitrogen (%) 8,09 2,99 0,279 C/N ratio 12,5

The chemical characteristics of the natural biodegradable product ECOSOL used in the experiment are presented in table 2. Table 2 Chemical characteristics of the natural biodegradable product ECOSOL Natural biodegradable product ECOSOL Total Nitrogen (%) 0,935 Organic Carbon (%) 23,72 Phosphorous Potassium (%) (%) 0,39 3,32 Sodium (%) 4,97

The inoculum was developed from microorganisms that occur naturally in the soil with the addition of crude oil. The microorganisms use the petroleum hydrocarbons as a source of carbon. It was stimulated the growth of the microbial population by adding the fibers provided from celluloid wastes. The density of crude oil used in artificial pollution of soil is been calculated with the following formula:

The value obtained for the crude oil used in the experiment is: = 836,9 kg/m3 This value shows that is a light crude oil. In the first experimental year was observed an increase of biodegradability rate in accordance with the applied treatment, so the experiment continued. In the second experimental year was used a plant to observe the influence of crude oil on plant growth and also to enhance the bioremediation by phytoremediation. The plant used was maize. RESULTS AND DISCUSSIONS Petroleum fuel or crude oil contaminated sites have been characterized by the total of all the petroleum hydrocarbons, called total petroleum hydrocarbons (TPH). In this paper are presented the influence of crude oil on plant growth. In figure 1 are presented the maize plants (was choosed one repetition) in each experimental variant after one month from emergence. The figure shows that in all the plots were the soil was polluted with 10% crude oil, the plants havent rise up even the treatment was applied. In the plots were the soil was polluted with 5% the plants rise up with delay, the plant are at least two times lower than control, and differences between treatments can be observed after one month.

241

Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009

Figure 1 Maize plant in each experimental variant after one month from appearence

In table 3 are presented the characteristcs of plants after 5 months from seeding, in the second experimental year of bioremediation and the first one when a plant was used in experiment. The number of plant/plot, the height of plants, the number of leaves and the biomass are proportional with the treatment. As it can be observed the control presents values very high for all the parameters analysed, the plots with soil polluted with 5% crude oil have values lower then control and the plots with soil polluted with 10% crude oil have no value because the plants did not rise up.

242

Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009

Table 3 Evolution of plant biomass in all experimental variants and repetitions according with the applied treatment
No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 Experimental variant Repetition R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 Plants/plot (number) 4 5 5 4 4 4 5 5 5 3 4 5 5 4 5 5 5 5 Height (cm) 138 140 139 46 53 57 57 50 51 57 56 52 68 47 56 83 48 56 Leaves (number) 12 13 12 7 9 8 7 7 8 8 9 7 9 8 8 8 9 8 Biomass (g) 99 113 107 6 4 5 7 5 3,5 5 5 6,5 7 6,5 5 8 5 6,5 -

V1, control (unpolluted soil)

V2, polluted soil with 5% crude oil

V3, polluted soil with 10% crude oil V4, polluted soil with 5% crude oil + 50 g ECOSOL V5, polluted soil with 5% crude oil + 50 g ECOSOL + bacterial inoculum V6, polluted soil with 5% crude oil + 100 g ECOSOL V7, polluted soil with 5% crude oil + 100 g ECOSOL + bacterial inoculum V8, polluted soil with 10% crude oil + 100 g ECOSOL V9, polluted soil with 10% crude oil + 100 g ECOSOL + bacterial inoculum V10, polluted soil with 10% crude oil + 200 g ECOSOL V11, polluted soil with 10% crude oil + 200 g ECOSOL + bacterial inoculum

243

Analele Universitii din Craiova, seria Agricultur Montanologie Cadastru Vol. XXXIX 2009

From treatment point of view, can be analysed all the parameters for the soil polluted with 5% crude oil. The number of plant/plot, the height of plants, the number of leaves and the biomass are increasing with the treatments applied. CONCLUSIONS The experimental study leads to the following preliminary conclusions: In the plots were the soil was polluted with 5% the plants rise up with delay, the plant are at least two times lower than control, and differences between treatments can be observed after one month. In all the plots were the soil was polluted with 10% crude oil, the plants havent rise up even the treatment was applied. The number of plant/plot, the height of plants, the number of leaves and the biomas are proportional with the treatment. For all the parameters analysed the control presents values very high, the plots with soil polluted with 5% crude oil have values lower then control, proportionaly with the treatment apllied and the plots with soil polluted with 10% crude oil have no value because the plants did not rise up. REFERENCES Alexander, M., (1994), Biodegradation and bioremediation, Publishers Academic Press, Inc. California, USA. Bartha R (1986), Biotechnology of petroleum pollutant biodegradation, Microbiology Ecology 12:155-172. Pepper, Ian L., Gerba, Charles P., Brusseau, Mark L (1996), Pollution Science, Academic Press. Ru, C., Zarioiu, V., Creang, I., Petre Neonila, Kaszoni Eniko, Crstea Stelian, Mihalache Gabriela, (1986). Cercetri preliminare privind tehnologia de redare n circuitul agricol a unor soluri poluate cu petrol, Analele ICPA, vol. XLVIII. Toti M., Dumitru, M., Cpitanu, V., Maria Dracea, Carolina Constantin, Constantin Crciun, (1999). Poluarea cu petrol i ap srat a solurilor din Romnia, Ed. RISOPRINT, Cluj-Napoca, 254 pag. Toti, M., Dumitru, M., Anca Voiculescu, M. Mihalache, Gabriela Mihalache, Carolina Constantin, (2003), Metodologia de bioremediere a solurilor poluate cu iei cu ajutorul mocroorganismelor specifice selecionate din micriflora autohton. I.S.B.N 973 85135 7 X; Ed. GNP Minischool, Bucureti. Voiculescu Anca Rovena, Gabriela Mihalache, M. Dumitru, M. Toti, (2002). Microbiological aspects on soil polluted Albic Luvisol under different remediation measures Albota Arge. International Conference Soils under Global Change a Challenge for the 21st Century. Constana Romnia, 3 6 septembrie.

244