Journal of Applied and Environmental Sciences 2008. Vol. 4, No. 2. pp.

118-124
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Effects of shooting range activities on heavy meals concentrations and

microbial activities in Nigeria soil environment

*1
Nwaugo, V.O. 2Ekpo, M.A., 2Eduok, S.I. and 3Verla, A.W.
1
Department of Microbiology, Abia State University, Uturu, Nigeria. 2Department of Microbiology,
University of Uyo, Nigeria. 3Department of Chemistry, Madonna University, Elele Campus, Nigeria

Received 17 September, 2008; Revised 3 December, 2008; Accepted 18 December, 2008

ABSTRACT

The soil physicochemical and biological parameter in Shooting Range Soil, near Port - Harcourt, Nigeria was
investigated. The concentrations of the heavy metals, Zn, Pb, Cu and Cd were highest in the soil at the Range
centre, followed by the midway soil sample before the edge. There was no remarkable change in pH, but temperature
followed the pattern of heavy metal concentration with acidic (lowest) values at the centre. Soil organic matter,
SO4, PO4, and NO3 negatively correlated with heavy metal concentration. Soil enzymes, dehydrogenase, urease,
phenol oxidase and acid phosphatase were adversely affected by the heavy metals. Dehydrogenase with a range
of 0.79- 8.24 mg g-1 dry soil 6h-1 was the least affected while acid phosphatase (0.42-2.11 mol-p-nitrophenol) was
the most affected. urease and phenol oxidase had ranges of 0.62-3.41 mg g-1 3h-1 and 0.57- 2.41 mg g-1 24h-1
respectively. Total Aerobic Heterotrophic bacteria had (4.7 x 10 3-4.4x106 cfu/g), Phoshatae solublizing bacteria
(0.4x101-2.1x104 cfu/g) and Nitrifying bacteria (0.3x101-3.4x103 cfu/g) were lowest at the range centre soil. This
shows that all the bacterial groups were affected with Nitrifying bacteria being the most affected. Results obtained
suggest the need for extensive rehabilitation and remediation, if the soil must be used for other human related
purposes such as farming or residence. Containment of the effects should be encouraged as soil immediately
outside the direct range activities was slightly affected probably due to rain water runoff effects.

Keywords: Enzymatic activities, soil shots, weapons, bacteria, heavy metal.

INTRODUCTION (2,4,6-Trinitrotoluene) and associated nitroaromatic

Activities at any shooting range cause the release
of several heavy metals and chemicals into the
environment. Detonation of weapons and des-
truction of expired or contaminated shots and
bombs affect the ecological balance and soil biota
(1). The high concentrations of heavy metals in
soil affect both the soil organisms and enzymes
adversely resulting in poor mineralization and
humification (2,3,4). This consequently leads to low
soil nutrients level, poor plant growth and migration
of soil dwelling annelids (5).
In most Military Shooting ranges, not only heavy
metals are released, toxic chemicals such as TNT
*
Corresponding Author
E-mai l: vonwango@yahoo.com
Tel .: +234 806 349 4654
substances are equally discharged into the
environment. According to Fuller and Mainin (6)
and Parmelee et al. (7), these chemicals affect
soil macro and micro flora and fauna adversely. It
has been reported that soil enzymes especially
dehydrogenase are equally adversely affected
(1,8).
These disturbances expose soil micro and macro
organisms to both heavy metal and chemical
intoxication, which invariably result in low
biodiversity in the affected soil.
Shooting ranges are areas mapped out, having
boundaries with adjoining areas, used for other
purposes. It has been reported that soil pollutants
are often spread to other areas not directly affected
by the initial pollution process due to rain water
run offs and flooding (4,9,10). The effects of these

118
Nwaugo et al.
range activities could therefore overlap to areas
not directly enclosed in the range.
Generally, shooting range soil conditions are not
often evaluated as the portion of land is often
considered a restricted area. This notwithstanding,
land is convertible, as new or larger ranges could
be acquired, leaving the former for other uses
including farming, and residential purposes. This
exposes the new users and organisms to the effects
of these heavy metals and chemicals available in
the soil.
This study therefore investigated the soil quality
in Igwuruta Shooting range near Port-Harcourt,
Rivers State Nigeria, using physicochemical and
biological parameters. This was aimed at assessing
the effects of shooting range activities on the soil
quality and the possibilities of using the land for
agricultural or residential purposes later.
MATERIALS AND METHODS
The study area
The study area is a shooting range near Port-
Harcourt, which has served the purpose for several
years over twenty five (25) years. It is located in
the typical rainforest region of Nigeria with its
characteristic tropical rain forest climate. The area
is covered by vegetation, the chief shrub being
Chromolena odoratum and some small trees of
assorted species.
Soil Analysis
The soil samples used in this study were ran-
domly collected from four different zones. The first
samples were collected around the centre of the
range, the second at the middle while the third was
at the edge. A fourth sample was collected from
5-6 meters outside the range while the control
sample was collected over 500m, well away from
the effects of the range.
Three samples were randomly collected from
each zone and pooled to give one main sample for
the zone. Sterile sample bottles were used for
collection of samples for microbiological and
enzyme analysis. Clean 250m amber reagent
bottles soaked in acid alcohol for 24 hours and
rinsed thrice in deionized water were used for
119
Journal of Applied and Environmental Sciences
samples collection for the physicochemical and
heavy metal analysis. All samples were collected
between June-September, 2007. The microbiolo-
gical and enzyme analysis were done within 2-4
hours of collection but the physicochemical and
heavy metals were accessed 1-3 days of collection.
Physicochemical and metal analysis
The soil temperature was determined in situ
using Mercury bulb thermometer (Jobling
laboratory Division, Stone, Staffordshire, ST 15
OBG, England) inserted into the soil. It was allowed
for 5-10 minutes before reading was taken. The
soil pH was determined using Jenway multipurpose
tester HANNA 1910 model (HANNA
Instruments, 584 Part East Drive Woonsuket,
Rhodes Island, USA). The phosphate, sulphate and
nitrate were determined by the procedures stated
in UNEP analytical methods for environment (11).
The organic matter content was determined by the
loss of ignition method using furnace (Jobling
laboratory Division, Stone, Staffordshire, ST 15
OBG, England) while the moisture content was
determined by the weight loss after drying to a
constant weight.
Soil enzyme analysis
The activities of enzymes determined include
dehydrogenase, urease, phenol oxidese, and the
phosphatases (acid). The dehydrogenase was
determined as described by Li et al (3) involving
the incubation of the soil sample with aqueous TTC
at 30oC. The absorbance of the triphenyl formazon
(TPF) produced was measured at 485nm after
methanol extraction. The result was expressed as
mg g-1 dry soil 6h -1. The urease activity was
determined by the colorimetric method of Kandeler
and Gerber (12). This method is based on formation
of NH3-N in urea-amended soil samples after 24
hours incubation at 37oC and expressed as mg
NH 3-N -1 dry soil 24h -1 . Phenol oxidase was
determined as described by Nannipieri et al. (13)
based on the formation of purpuragallin formation
in pyrogallic acid-amended soil after incubation for
3 hours at 30 oC. The result was expressed as
mg-1 dry soil 3h-1. Acid phosphatase activities were
determined as described by Tabatabai (14) using
p-nitrophenyl phosphate incubation with the soil
Vol. 4, No. 2. 2008 Soil quality and enzyme activities

sample and the result was expressed as mol-p-
nitrohenol.
Microbiological analysis
The bacteira diversity and population in each of
the sample soil were determined using various
culture media. The bacterial groups estimated were
the Total Aerobic heterotrophic Bacteria (TAHB),
the Nitrifying Bacteria (NB), and the Phosphate
Solubilizing Bacteria (PSB). The TAHB was
determined using Tryptone Soy Agar, NB with
Mineral Salt Agar and PSB with Phosphate
Solubilzing Bacteria Medium as outlined in US
Patent 6638730 (15). The soil samples were
inoculated on te various media using the spread
plate technique.
Analysis of data
The results obtained in this study were subjected
to statistical analysis using two main methods -
Correlation analysis for the enzymes activities with
heavy metal concentrations. This was also used
for the bacterial loads and heavy metal
concentrations. The heavy metals and physico-
chemical parameters in soil sampling areas were
compared using the standard deviation to test for
the differences.
RESULTS
The results of the physicochemical parameters
of the various soil samples are shown in Table 1.
The soil temperature showed that values at the
Shooting range centre and midway were
statistically different from the other areas sampled
while that other areas were similar to each other.
The pH which ranged between 6.21 to 7.62 and
moisture content followed the same pattern with
temperature. Soil organic matter, SO4, PO4 and
NO3 followed a gradient with lowest values at the
range centre and gradually increased till the edge.
The 5-6m outside the range sample value was quite
close to the control. Organic matter content ranged
between 4.22-8.37 mg/g, NO3 was 0.52 - 3.91 mg/
g while SO4 and PO4 had 2.12-4.82 mg/g and 16.37
- 29.78 mg/g ranges respectively (Table 1).In each
case, highest values were in the control while the
least were at the range centre soil samples.
The heavy metal concentrations increased with
distance to the centre of the range. All the heavy
metals had their highest concentrations in the range
centre soil sample, followed by the midway soil
sample before the edge sample. The values of soil
samples collected from 5-6m outside the edge
were close to those of the control. Zn had the
highest concentration in the soil with values ranging
from 44.31 to 214.61 mg/g, followed by Pb (41.61-
183.14 mg/g) and Cu, 41.54-172.31 mg/g. Cd was
the lowest (0.02-0.97 mg/g). The difference of
the metal concentrations according to sampling
area were statistically significant (P=0.05) (Table
2).
Table 3 shows the values of soil enzymatic
activities which correlated negatively with the
heavy metals concentrations. Soil samples with high
heavy metal concentrations showed low enzymatic
activities while those with low heavy metal
concentrations had higher soil enzymatic activities.
All the enzymes had significantly different activity
values at each sampling point.
Among the soil enzymes, dehydrogenase had the
highest activities with a range of 0.79-8.24 mg g-
1
dry soil 6h-1, followed by urease with 0.62-3.41

Table 1. Physicochemical parameters of the soil samples

Parameters Physicochemical parameters of the soil

Center Midway Edge Outside Fence Control
a a b b b
Temp. 29.3+0.34 29.1+0.41 27.2+0.12 27.0+0.21 26.8+0.16
a a b b c
pH 6.21+0.41 6.24+0.43 6.87+0.61 7.01+0.40 7.62+0.71
M oisture content 15.7+0.21a 16.2+0.27a 17.30.40b 18.9+0.62c 18.31+0.64c
SOM mg/g 4.22+0.31a 4.77+0.34a 5/ 71g+0.52b 6.14+0.80c 8.37+0.81d
SO4 gg/g 2.12+0.11a 2.92+0.14a 3.11+0.14a 4.01+0.42b 4.82+0.64c
PO4 mg/ g 16.37+0.41a 17.81+0.47a 19.27+0.24b 21.43+0.26b 29,78+0.67c
a b c c
NO3 mg/g 0.52+0.112 0.84+0.20a 1.20g+0.42 2.12+0.61 3.8+0.82
*Figures followed by the same alphabets are not significantly different but those followed b y different alphabets are significantly different.

120
Nwaugo et al. Journal of Applied and Environmental Sciences

mg g-1 24h-1. The least acting enzyme was acid DISCUSSION

phosphatase with a range of activities of 0.42-2.11
mg g-1 3h-1 (Table ). Phenol oxidase had activity in Results obtained in this study suggest that the
the range of 0.57-2.41 mg g-1 6h-1. activities at the shooting range significantly
Table 2. Concentratio ns of heavy metals observ ed in th e shooting range s oil

Heavy Physicochemical parameters of the soil

metals Center Midway Edge Outside Fence Control
Zn mg/g 214.61+0.11a 186.61+0.21b 162.42+0.36c 46.1+1.2d 31.31+1.6d
Pb mg/g 183.14+0.16a 163.67+0.20 a
112.45+0.42 b
42.31+1.40c 20.61+1.62d
Cu m g/g 172.31+0.61a 126.54+0.87b 107.25+0.91c 41.11+1.41d 29.54+1.68e
Cd m g/g 0.97+0.11a 0.54+0.23 b 0.21+0.43c 0.04+0.68d 0.01+0.60d
*Figures followed by the s ame alph abets are not s ignificantly d ifferen t but those follow ed by d iffer ent alphab ets are s ignificantly
different.

Ta ble 3. Enz yma tic ac tivit ies of the soil

Enz ymes P hysicochemical pa rame ters of the soil

Ce nter Midwa y Edge Outside Fenc e Control
Dehydrogenase
-1 -1 a b c d c
mg g dry soil 6h 0.79+0.21 1.82+0.43 3.60+ 0.67 6.78+0.88 8.24+ 0.92
Urea se
mg g -1 24 h -1 0.62+0.21 a 0.93+0.40 b 1.21+ 0.59 c 2.42+0.72 d 3.41+ 0.96 e
Aci d phoshat ase
-1 -1 a b c d e
mg g 3h 0.42+0.22 0.82+0.27 1.21+ 0.40 1.87+0.61 2.11+ 0.82
P henol oxi dase
mg g -1 6h -1 0.57+0.13 a 0.72+0.42 b 1.11+ 0.67 c 1.48+0.83 d 2.41+ 0.87 d
*Figures follow ed by the s am e alpha bets are not s igni fica ntly diffe rent but those fol lowed by different alphabets are signific ant ly
different.

The bioloads of the various groups of organisms affected the soil quality when compared to the
(Table 4) indicates that the most abundant bacterial control. Concentrations of the heavy metals at the
group was the total aerobic heterotrophic bacteria, range were statistically higher than those of the
followed by the phosphate solubilizing bacteria control soil. According to CEC directive (18), values
while the nitrifying bacteria were the least. In each of Pb, Cu, Cd and Zn obtained in the study were
case, highest bioload was reported in the control. too high. Values obtained 5-6m outside the range
The soil sample 5-6m outside the range had values and control further buttressed this observation by
next to the control while the least values were at being lower than those inside the shooting range.
the Shooting Range centre. TAHB counts ranged The high concentrations of heavy metals at the
from 4.7x103 cfu/g at the centre, through 6.7 x 103 centre and midway of the range (study area) could
cfu/g (midway), to 1.5 x 104 at the edge. The be due high activities involving weapon discharge.
control had 4.4x106 cfu/g. PSB had a range of These ammunitions contained Cu, Pb and Zn alloys,
0.41x101-2.7x104 cfu/g (highest at control, lowest which could add to the concentrations in the soil.
at centre). The NB had significantly lower values These additions resulted in the increase in values
with 0.3x10 1 cfu/g at the centre, followed by above those from the samples from the shooting
3.1x101 at the middle, and 1.4x102 at the edge. range edge and control. Lee et al. (1) had stated
The control had 3.4 x 103 cfu/g for NB. Statistical that normal activities at any shooting range result
analysis showed significant difference (P=0.05) in in the release of heavy metals into the environment
the bioloads. as observed in this study. The high concentrations
Table 4. Est imated bi olaod of various bacterial groups(cfu/g)

Bacterial C ent er Mi dway Edge ???? C ontrol

group
3 a 3 b 4 c 6 d
TAHBC 4.7X10 +0.31 6.7X10 +0.49 1.5X10 +0.63 2.1X10 +0.83 4.4X106 +0.93c
PSBC 0.4X101 +0.12 a 3.1X102 +0.31 b 2.6X10 3+0.49c 1.9X10 4 +0.72d 2.7X104 +0.70f
NB C 0.3X101 +0.16 a 3.1X101 +0.02 b 1.4X10 2+1.60c 1.1X10 3 +2.11d 3.4X103 +2.43e
*Figures follow ed by the s ame alphabets are not s ignificantly different but those follow ed by different alphabe ts are s ignifi cantly differe nt.
T AHBC Total Aerobic He terotrophic Bacteria l Count; PSBC Phos phate Solubilizing Bacterial Count; NBC Nitrifying Bacterial Count

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Vol. 4, No. 2. 2008
of these metals in the soil immediately out side the
range, could be due to rain water runoff which
carry away some of the metals from the point of
pollution. Nwaugo et al. (10, 17) and Chinyere (9)
had earlier reported similar observations as pollutant
concentrations form a gradient away from the point
of pollution.
Observations of the physicochemical parameters
suggest adverse effects on soil quality through the
shooting activities too. While the most affected
areas were slightly acidic, soil nutrients such as
PO4, NO3 and SO4 including organic matter were
equally low in these highly contaminated areas.
These suggest antimineralization and anti humifi-
cation effects on the soil which are unlike the situ-
ation in the control sample and samples of soil out-
side the shooting range. These observations agree
well with those of Lee et al. (18) in South Korea.
Lee et al. (1) also reported that high concentra-
tions of heavy metal reduce organic matter. These
observations agree with Prescott et al. (19) and
Pelczar et al. (5). This study therefore agrees that
high heavy metal concentrations reduce soil nutri-
ent levels, soil moisture and pH.
The results obtained on the enzymatic activities
confirmed earlier findings that heavy metals affect
soil enzymes adversely (3, 4, 8, 20). In all the
enzymatic activities estimated, the results
correlated positively with the bioloads of microbial
groups estimated as reported elsewhere. Heavy
metals tend to inhibit the activities of these enzymes
including other processes in the microorganisms
resulting in low enzymatic activities and microbial
population in contaminated soil.
The most affected enzyme was dehydrogenase.
The activity of the enzyme at the shooting range
centre was only 9.58% of the control value. The
middle had only 14.56 % of the control value too.
Similarly the urease value at the shooting range
was was only 18.18 % of the control value. The
low urease activity observed in this study agree
with the report of Prescott et al., (19) that nitro-
aromatic compounds, as could be found in some
military weapons, could inhibit nitrogen compound
mineralization. Urease is involved in nitrogen
contents of the soil. Kuperman and Carreiro (21)
agreed that high microbial loads lead to high de-
122
Soil quality and enzyme activities
dehydrogenase and phosphatase activities. This
observation could be early seen in this study. These
are enzymes produced by the microorganisms, it
therefore follows that the more the bacterial
population, the higher the amount of enzymes
released hence the higher the activity. The opposite
applies true for low microbial density soil as
reported in the shooting range centre.
Observation of the various bacterial groups
shows that the most affected organisms were the
nitrifying bacteria. Nwaugo et al. (22) and Prescott
et al. (19) stated that nitrifying bacteria are very
sensitive to environmental disturbances. Nwaugo
et al. (10) agree with Lee et al. (1) that high
nitrogen compound in the soil will suppress soil
nitrification process. This could be the case, as
nitro aromatics form the major components of
military chemicals such as TNT used in Shooting
Ranges. PSB were equally affected adversely
along with Aerobic heterotrophic bacteria. The
AHB are the sum total of all culturable aerobic
bacteria in the soil, hence they include even the
phosphate solubilzing bacteria and some nitrifying
bacteria capable of growing on the Tryptone Soy
Agar used. The bioloads of these organisms
positively correlated with the enzymatic activities,
as the higher the bioload, the higher the enzymatic
activities.
The results obtained in this study therefore
suggest that the Shooting range soil lacks soil
nutrients following reduced microbial and
enzymatic activities. This study then suggests that,
if the soil could be used for any other purpose, the
heavy metal level most be reduced and the soil
rehabilitated. The rehabilitation becomes very
essential, if the soil will be put into agricutlrual uses
because most of the plants will bio accumulate
these metals in their tissues.
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