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Meri-Rastilantie 3 C, FI-00980 Journal of Food, Agriculture & Environment Vol.4 (3&4) : 218-222. 2006
Helsinki, Finland

Comparison of corn and elephant grass in the phytoremediation of a petroleum-

hydrocarbon-contaminated agricultural soil in Port Harcourt, Nigeria

Josiah M. Ayotamuno *, Reginald B. Kogbara and Paul N. Egwuenum

Agricultural & Environmental Engineering Department, Rivers State University of Science & Technology, Port Harcourt, P.M.B.
5080, Rivers State, Nigeria. *e-mail:,,

Received 29 May 2006, accepted 3 September 2006.

Two types of plants, Zea mays (corn) and Pennisetum purpureum (elephant grass) were compared in field studies to evaluate their potential in
degrading petroleum hydrocarbons in contaminated agricultural soils. Previous studies have shown that certain plants together with their associated
microorganisms could increase the removal of petroleum hydrocarbons from contaminated soils. Consequently, field plots comprising a control and
five treatment plots, whose treatment options entailed the use of corn, elephant grass and corn plus elephant grass, were utilized in the research
design. The experimental methodology involved the simulation of conditions of a major spill through the sprinkling of crude oil on the plots, after
which the plants were introduced to the plots. The Z. mays seeds were first grown in nurseries before they were transferred to the plots, while mature
already grown P. purpureum stands were transplanted on the experimental plots. Fertilizer application followed thereafter. Relevant soil physicochemical
parameters and bacterial counts were analyzed and monitored during the study period. The results of the analyses revealed average hydrocarbon
losses of 77.5% (Z. mays) and 83% (P. purpureum) within the first two weeks, these values decreased to 67.5% and 55% after the six-week
remediation period for corn and elephant grass respectively. On the other hand, the corn plus elephant grass treatment showed hydrocarbon losses
of 62% and 74% for the two and six-week period respectively. These values differed greatly from those obtained in the control plot. It is evident from
the percentage hydrocarbon losses recorded in this study that over a wide range of field conditions, corn degrades the contaminant better than elephant
grass and the combination of both plants yields the best alternative in the phytoremediation of a petroleum-hydrocarbon-contaminated agricultural
soil. The findings of this research further highlighted the position that with ample time at the disposal of stakeholders, remediation of petroleum-
hydrocarbon-polluted agricultural soil using corn and elephant grass treatments yields a cost-effective and successful approach. What is necessary for
a remarkable remediation to be achieved is the creation of favorable conditions within the soil environment, like adequate nutrient supplementation
and oxygen availability, for the proper development of soil microbes and plant utilization of the contaminant for their metabolism.

Key words: Petroleum contamination, phytoremediation, Pennisetum purpureum, Zea mays, phytoaccumulation, phytovolatilization, biodegradation,
total hydrocarbon content, hydrocarbon utilizing bacteria, soil nutrients.

Contamination of soils with petroleum hydrocarbons is one of the contaminated soil is an increase in microbial activity. Supporting
attendant environmental problems associated with this hypothesis, the population levels of contaminant degrading
industrialization and the dependence on petroleum and its by- bacteria and the potential of the soil to degrade contaminants
products. Consequently, several technologies have been resorted typically increase during phytoremediation 5, 6. Phytoaccumulation
to in a bid to effect the clean up of contaminated soils. One of (the uptake of hydrocarbons from contaminated soils by plants)
such technology is phytoremediation, which entails the in situ and phytovolatilization (the use of plants to transfer volatile
use of plants and their associated microorganisms to degrade, petroleum hydrocarbons from the soil to the atmosphere) could
contain or render harmless contaminants in soil 1. Various plants be the mechanisms responsible for phytoremediation 7, 8. From the
have been identified for their potential to facilitate the foregoing it can be deduced that whatever the mechanism involved
phytoremediation of sites contaminated with petroleum in phytoremediation the technology is a cost effective alternative
hydrocarbons. In majority of the studies grasses and legumes to conventional remediation methods. However, the
were singled out for their potential in this regard 2. However, the phytoremediation option is relatively new in the tropics, thus there
mechanisms involved remain poorly understood. is a dearth of information in this area. Sequel to the above, this
Studies carried out by Lee and Banks 3 revealed that the plant study became pertinent in Port Harcourt, an important city of the
root zone (rhizosphere) has significantly larger numbers of Niger Delta region of Nigeria, where incessant oil spills from oil
microorganisms than soils, which do not have plants growing in exploration and development activities have devastated farmlands
them; this appears to enhance the biodegradation of organic
compounds. In a related view, Siciliano et al. 4 hypothesized that Nomenclature: THC total hydrocarbon content mg/kg, EC electrical conductivity, CH4 methane, OC
the mechanism responsible for the phytoremediation of organic carbon, TN total nitrogen, cfu/ml colony forming units per millilitre, HUB hydrocarbon utilizing

218 Journal of Food, Agriculture & Environment, Vol.4 (3&4), July-October 2006
and other agricultural settlements over the years 9. mays seedlings, which were already been grown in nursery beds,
In this study we assessed the potential of phytoremediation to were transplanted on the plots using a hand trowel, while already
degrade petroleum hydrocarbons, especially in tropical soils. growing P. purpureum within the experimental area were transferred
Hence Z. mays and P. purpureum, which are common tropical to the plots. Each of the plots contained five stands of plants.
plants, were utilized for phytoremediation treatment. The potential Soil samples were collected at intervals for laboratory analysis.
of the indigenous soil microbial flora associated with these plants, This was done using a hand dug soil auger. The soil was carefully
to mineralize petroleum hydrocarbons, was also assessed. augered to 30 cm depth at different spots and bulked together to
Therefore, the objectives of this study were to determine if form composite soil samples such that no harm was inflicted on
phytoremediation treatments involving corn and elephant grass the plants. This procedure was done three times to form three
could facilitate the quick mineralization of petroleum hydrocarbons replicates. The samples were put in well-labeled polyethylene bags
and to compare the phytoremediation potential of the individual and transferred to the laboratory for analyses.
plants and also a combination of both plants, in the degradation
process, with the view of ascertaining the alternative that yields a Laboratory/statistical methods: Relevant soil physicochemical
better option for a tropical soil. parameters, such as particle size composition, total hydrocarbon
content (THC), organic carbon, total nitrogen, moisture content,
Materials and Methods pH and electrical conductivity were analyzed using methods
Study area description: The experimental plots were located at adapted from Black et al. 11. Microbiological analyses were carried
the University Teaching and Research Farm in the Rivers State out following the procedure described by the American Public
University of Science and Technology, Port Harcourt, Nigeria. Health Association (APHA) 12 and Harrigan and McCane 13.
Port Harcourt is an important city in the Niger Delta region of Univariate summary statistics such as mean, standard deviation
Nigeria. The Niger Delta region produces over 98% of Nigerias and percentage, and correlation and regression analysis 14 were
economic mainstay, crude oil. The region lies between latitudes employed to analyze data. The correlation and regression analysis
415N and 630N and longitudes 500E and 800E. Port Harcourt was done to determine the relationship between time (in days)
falls within the tropical rain forest vegetation belt of the country and measured soil characteristics across the plots, since the
and receives an annual rainfall of about 2700 mm. The average technology under study is a time dependent process.
temperature of the experimental area is about 27C.
Results and Discussion
Field sites: The experimental design involved six field plots that Tables 1-4 show the relevant physicochemical characteristics of
were made into beds each with dimensions 40 cm x 40 cm, and the the soil that were analyzed and used as indices for evaluation of
beds were about 30 cm deep. The primary functions of the beds the pollution/remediation process. The particle size analyses of
were to control the exposed surface area of the soil, the nutrient the top 30 cm of the soil before treatment indicated that the soil
concentration, temperature and oxygen availability 10. They also texture is silty clay (Table 1). There was general increase in soil
served to prevent excessive runoff of the contaminant, which is a moisture content in most of the plots during the study period
common trend in the locality during the period the study was (Tables 14). This was expected due to the large volume of rainfall
conducted (June to August, 2005) sequel to the large volume of that characterized the study period, which traversed the month of
rainfall and the nature of the soil. Plot O was the control site (with July, one of the rainfall maxima in the country. There was a
no treatment employed), plots B and E received elephant grass correlation (r = + 0.041) between soil moisture content and
treatment, plots C and D received corn treatment, while plot A had remediation period, which was not significant at 5% probability
a mixture of corn and elephant grass growing on it. level (Table 5). This follows from the fact that moisture variations
in the various plots were not significant with progress in
Experimental procedure: The plots were first tilled to 30 cm depth remediation period.
to allow for easy penetration of plant roots and infiltration of the Soil pH increased in the aftermath of crude oil contamination
crude oil contaminant. Thereafter 800 cm3 (0.8 litres) of Bonny (Tables 1 and 2). It later dropped during the course of remediation
light crude oil was applied to each plot by sprinkling from treatment (Tables 3 and 4). This may have been due to the
perforated cans. The objective was to simulate conditions of a production of acid radicals during the process of nitrification of
major spill. The plots were left undisturbed for three days. After the applied fertilizer. Tisdale and Nelson 15 made a similar
three days, 50 g of 20-10-10 NPK fertilizer was applied in solution observation. There was no significant decrease in the effect of
to the treatment plots in order to provide nitrogen, which is a the remediation on soil pH (r = - 0.060) at 5% probability level
major limiting nutrient in the growth of soil biota. The same quantity (Table 5). The electrical conductivity (EC) of the soil followed a
of fertilizer was applied again after three weeks of remediation similar trend but in this case there was a general increase in the EC
treatment. In essence, all the plots received 1250 kg ha-1 of nitrogen of most of the treatment plots during the remediation period (Tables
and 625 kg ha-1 of phosphorus and potassium, throughout the 1-4). This must have been due to the soluble salt content in the
remediation period. In the aftermath of fertilizer application, the Z. soil induced by the introduction of the inorganic fertilizer.

Table 1. Soil physicochemical characteristics before crude oil contamination (mean+standard deviation of treatment plots).
% by weight pH EC THC % by weight C / N ratio
Sand Silt Clay Moisture 1: 2.5 S cm-1 mg kg-1 Organic C Total N
12.4+0.3 40.1+0.2 47.5+ 0.7 13 + 1 4.71 + 0.25 45 +2 25+10 0.19+0.02 0.38 + 0.03 0.5 +0.02

Journal of Food, Agriculture & Environment, Vol.4 (3&4), July-October 2006 219
The organic carbon content of the soil increased after activities of denitrifying bacteria such as Pseudomonas, Bacillus
contamination. This was caused by the introduction of large and Micrococcus, especially when localized micro sites of low
amounts of hydrocarbons into the soil, which buttresses the oxygen exist in the center of soil aggregates. These microbes
position of Jobson et al. 16 that oil spills result in significant were isolated from the soil samples during microbiological
increase in percent organic carbon because crude oil is essentially analyses; hence they may have induced the trend. The continuous
a mixture of carbon and hydrogen. However, a major effect of the utilization of available nitrogen implied that the nitrogenous
remediation treatment(s) was the enormous attenuation of the nutrient supplied favored biodegradation. The correlation analysis
organic carbon content of the treatment plots (Tables 2-4). There between total nitrogen and remediation period yielded a negative
was a correlation (r = -0.251) at 5% probability level between organic correlation (- 0.427), which was significant at 5% probability level
carbon and remediation period (Table 5). This suggests that (Table 5) indicating the rapid utilization of the soil nutrient by soil
organic carbon reduced with time. On the other hand, the total biota across the various plots. Microbiological analysis of the
nitrogen decreased throughout the study period amidst the soil samples showed that the hydrocarbon-utilizing bacteria (HUB)
application of the nitrogenous fertilizer. This is unusual and associated with the plants include Pseudomonas, Bacillus,
contrary to expectation that the total nitrogen would naturally Micrococcus, Corynebacterium, Flavobacterium, Acinetobacter,
increase with the application of the nitrogenous fertilizer. That Alcaligenes and Chromobacterium. In the course of the study
not withstanding, such a result is not utterly strange as a related the hypothesis that an increase in microbial activity was the
study carried out during the same period in the same experimental mechanism responsible for phytoremediation was substantiated
area 17 witnessed the same situation. The high levels of bacterial by the enormous increase in bacterial counts of the treatment
activity in the soil must have induced the trend. FitzPatrick 18 plots (Table 6). The trend was different for the control site, which
reported that in addition to biological uptake nitrogen could be had no plant growing on it. Its total heterotrophic bacteria first
rapidly lost from soils by leaching of ammonia and by denitrification decreased from 22x105 to 19.8x105 cfu ml-1 and later increased to
in wet soils. Brady and Weil 19 have also reported a similar 25.4x105 cfu ml-1 (Table 6). This highlights the view that the
observation. The authors noted that during biodegradation enormous increase in bacterial count in the treatment plots is
nitrogen may be lost to the atmosphere when nitrate ions are linked with the treatment applications (i.e. planting of corn and
converted to gaseous forms of nitrogen by a series of widely grass plus nutrient supplementation). Similarly, the view that
occurring biochemical reduction reactions induced by the phytovolatilization is a likely mechanism of phytoremediation
Table 2. Soil physicochemical characteristics 3 days after contamination, prior to remediation (mean+
standard deviation of three replicates).
Plot Moisture % pH EC THC % by weight C/N
by weight 1: 2.5 S cm-1 mg kg-1 Organic C Total N ratio

O 10 + 2 6.05+ 0.30 52 + 4 18,145 + 20 0.440 + 0.05 0.210 + 0.02 2 + 0.5

A 12 + 1 6.15 + 0.19 89 + 3 20,150 + 50 0.460 + 0.03 0.190 + 0.01 2 + 0.3
B 18 + 1 6.04+ 0.25 96 + 3 24,917 + 120 0.500 + 0.04 0.200 + 0.02 3 + 0.6
C 16 + 2 6.02+ 0.20 98 + 2 24,683 + 240 0.480 + 0.05 0.220 + 0.03 2 + 0.4
D 14 + 1 6.08+ 0.30 99 + 5 20,470 + 100 0.500 + 0.032 0.310 + 0.01 2 + 0.2
E 16 + 1 6.04+ 0.20 103 + 4 22,086 + 250 0.500 + 0.04 0.330 + 0.04 2 + 0.5

Table 3. Soil physicochemical characteristics 2 weeks after remediation (mean+standard deviation of three
Plot Moisture % pH EC THC % by weight C/N
by weight 1: 2.5 S cm-1 mg kg-1 Organic C Total N ratio

O 10 + 2 5.64+ 0.20 45 + 7 12, 010 + 60 0.380 + 0.02 0.170 + 0.08 2 + 0.5

A 16 + 1 5.52 +0.05 99 + 9 7,661 + 110 0.330 + 0.03 0.123 + 0.007 3 + 0.6
B 16 + 1 5.55 + 0.20 91 + 6 5,004 + 250 0.390 + 0.02 0.126 + 0.004 3 + 0.6
C 15 + 1 5.35 + 0.20 102 + 5 5,542 + 200 0.380 + 0.05 0.139 + 0.006 3 +0.5
D 15 + 2 5.26 + 0.30 90 + 4 4,730 + 180 0.440 + 0.01 0.194 + 0.007 2 + 0.6
E 22 + 1 5.45 + 0.10 124 + 15 2,985 + 210 0.400 + 0.06 0.201 + 0.006 3 + 0.4

Table 4. Soil physicochemical characteristics 6 weeks after remediation (means+standard deviation of

three replicates).
Plot Moisture pH EC THC % by weight C/N
% by weight 1: 2.5 S cm-1 mg kg-1 Organic C Total N ratio

O 14 + 2 5.44 + 0.30 39 + 5 20,414 + 90 0.340 + 0.04 0.148 + 0.004 2 + 0.8

A 17 + 3 5.18 + 0.20 102 + 6 5,239 + 30 0.190 + 0.05 0.057 + 0.003 3 + 0.7
B 16 + 1 5.50 + 0.30 95 + 9 11,223 + 80 0.280 + 0.05 0.056 + 0.002 5 + 0.8
C 19 + 2 5.11 + 0.20 104 + 8 8,502 + 200 0.260 + 0.06 0.061 + 0.0002 4 + 0.6
D 16 + 1 5.05 + 0.30 97 + 6 6,363 +220 0.240 + 0.05 0.081 + 0.004 3 + 0.5
E 17 + 2 5.21 + 0.30 256 + 12 9,936 + 200 0.300 + 0.07 0.084 + 0.005 4 + 0.6

220 Journal of Food, Agriculture & Environment, Vol.4 (3&4), July-October 2006
Table 5. The relationship between time and some measured soil characteristics
during remediation as expressed by correlation coefficient values and
regression equations.
Correlation r Significance Regression equation
time vs. moisture +0.041 ns Y = 15.85 + 0.026x
time vs. pH -0.060 ns Y = 5.81 + 0.013x
time vs. THC -0.346 ns Y = 15 511 191x
time vs. organic C -0.251 ns Y = 0.451 0.004x
time vs. total N -0.427 * Y = 0.214 0.003x
time vs. percentage THC reduction +0.452 * Y = 28 + 1.0x
*significant at 5%; ns not significant at 5% probability level.

Table 6. Total heterotrophic bacterial count. 100

Sampling period (weeks)

Percentage THC reduction

Plot 0 2 6 80
(x10 5 cfu ml-1) 70
O 22.0 19.8 25.4 60
A 20.4 28.8 38.6
B 8.95 20.4 33.6 50
C 8.65 17.0 28.4 40
D 21.1 22.4 31.6
E 20.5 24.9 38.2
* The background (i.e. pre-contaminated) samples had a bacterial count of 20
52x105 cfu/ml.
Table 7. THC reduction percentage.
Treatment Sampling period 0 2 6 8
plot (weeks) Weeks after remediation
2 6 Corn Elephant grass Corn plus Elephant grass
A 62 74
B 80 55 Figure 1. Average values of THC reduction percentage for the three kinds of
C 78 66 treatments.
D 77 69
E 86 55 of anoxic zones where degradation is dominated by anaerobic
microorganisms. The resulting anaerobic degradation must have
remains attractive. An evidence of this was the leaf burn (leaf given rise to methane (CH4) and carbon dioxide, the products of
chlorosis) observed in the plants during the first few weeks of anaerobic decomposition of organic materials. The gradual
remediation. Such a situation suggests that volatile organic formation of methane, a hydrocarbon, could most likely cause an
compounds were taken up by the roots of the plants, translocated increase in the total hydrocarbon content.
within the plants and transpired through the stems and leaves 7. The above hypothesis emanates from the fact that any increment
The leaf burn gradually disappeared with time indicating that many in THC must have resulted from within the soil environment since
of the volatile petroleum hydrocarbons had been transferred to the basic source of hydrocarbon, the crude oil, was applied just
the atmosphere. However, this may have implications regarding once during the study. The same trend was observed in a related
subsequent pollution of the ambient atmosphere. study carried out during the same period 17. From the study it was
The results of the total hydrocarbon content (THC) show that observed that field plots undergoing regular and optimum levels
phytoremediation could achieve the degradation of petroleum of tillage (and hence oxygen exposure) had very little or no traces
hydrocarbons. After two weeks of remediation the percentage of increment in contaminant concentration unlike the ones without
THC reduction for all the options was 62, 80, 78, 77 and 86 for regular tillage. Vance 20 observed and reported a similar trend of
plots A, B, C, D and E respectively (Table 7). The average values sudden increase in contaminant concentration. He posited that it
for corn- and grass-treated plots were 77.5 and 83% respectively, might be due to certain extracellular surfactant-like polymers
while that of corn plus grass treatment was 62% (Fig. 1). excreted by the hydrocarbon-utilizing microbes during their
Correlation and regression analysis indicated that the THC metabolic cycle. All in all, the exact mechanism responsible for
reduction percentage was significant with increase in remediation this trend remains not well understood. Statistical analysis of the
period at 5% probability level (Table 5). From the regression THC results indicated a weak correlation (- 0.346) with time, which
equation, it can be deduced that on the average, a 35% reduction was not significant at 5% probability level thus highlighting the
in THC can be obtained on a weekly basis (see Table 5, Column 4). fact that the THC of the plots actually decreased with time in the
Moreover, the sample set collected after six weeks of remediation long run but its variation with time also depended on other factors;
gave anomalous appearing results, as the THC of the plots were one of which was the moisture condition of the soil, and hence
generally higher than the ones obtained after the two week period the oxygen availability.
(Tables 3 and 4). The end of the six week period fell in early August, During the first two weeks the plots that received elephant grass
immediately after the rainfall maxima in July, hence it could be treatment generally thrived better than the corn-treated plots,
hypothesized that the high moisture content associated with that probably due to the ease of adaptation of elephant grass to the
period limited gas exchange in the soil and resulted in the creation contaminated soil by reason of its faster root development. As

Journal of Food, Agriculture & Environment, Vol.4 (3&4), July-October 2006 221
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222 Journal of Food, Agriculture & Environment, Vol.4 (3&4), July-October 2006