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

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Effects of shooting range activities on heavy meals concentrations and

microbial activities in Nigeria soil environment

Nwaugo, V.O. 2Ekpo, M.A., 2Eduok, S.I. and 3Verla, A.W.
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


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

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

Nwaugo et al. Journal of Applied and Environmental Sciences

range activities could therefore overlap to areas samples collection for the physicochemical and
not directly enclosed in the range. heavy metal analysis. All samples were collected
Generally, shooting range soil conditions are not between June-September, 2007. The microbiolo-
often evaluated as the portion of land is often gical and enzyme analysis were done within 2-4
considered a restricted area. This notwithstanding, hours of collection but the physicochemical and
land is convertible, as new or larger ranges could heavy metals were accessed 1-3 days of collection.
be acquired, leaving the former for other uses
including farming, and residential purposes. This Physicochemical and metal analysis
exposes the new users and organisms to the effects The soil temperature was determined in situ
of these heavy metals and chemicals available in using Mercury bulb thermometer (Jobling
the soil. laboratory Division, Stone, Staffordshire, ST 15
This study therefore investigated the soil quality OBG, England) inserted into the soil. It was allowed
in Igwuruta Shooting range near Port-Harcourt, for 5-10 minutes before reading was taken. The
Rivers State Nigeria, using physicochemical and soil pH was determined using Jenway multipurpose
biological parameters. This was aimed at assessing tester HANNA 1910 model (HANNA
the effects of shooting range activities on the soil Instruments, 584 Part East Drive Woonsuket,
quality and the possibilities of using the land for Rhodes Island, USA). The phosphate, sulphate and
agricultural or residential purposes later. nitrate were determined by the procedures stated
in UNEP analytical methods for environment (11).
MATERIALS AND METHODS The organic matter content was determined by the
loss of ignition method using furnace (Jobling
The study area laboratory Division, Stone, Staffordshire, ST 15
The study area is a shooting range near Port- OBG, England) while the moisture content was
Harcourt, which has served the purpose for several determined by the weight loss after drying to a
years over twenty five (25) years. It is located in constant weight.
the typical rainforest region of Nigeria with its
characteristic tropical rain forest climate. The area Soil enzyme analysis
is covered by vegetation, the chief shrub being The activities of enzymes determined include
dehydrogenase, urease, phenol oxidese, and the
Chromolena odoratum and some small trees of
phosphatases (acid). The dehydrogenase was
assorted species.
determined as described by Li et al (3) involving
the incubation of the soil sample with aqueous TTC
Soil Analysis
at 30oC. The absorbance of the triphenyl formazon
The soil samples used in this study were ran-
(TPF) produced was measured at 485nm after
domly collected from four different zones. The first
methanol extraction. The result was expressed as
samples were collected around the centre of the mg g-1 dry soil 6h -1. The urease activity was
range, the second at the middle while the third was determined by the colorimetric method of Kandeler
at the edge. A fourth sample was collected from and Gerber (12). This method is based on formation
5-6 meters outside the range while the control of NH3-N in urea-amended soil samples after 24
sample was collected over 500m, well away from hours incubation at 37oC and expressed as mg
the effects of the range. NH 3-N -1 dry soil 24h -1 . Phenol oxidase was
Three samples were randomly collected from determined as described by Nannipieri et al. (13)
each zone and pooled to give one main sample for based on the formation of purpuragallin formation
the zone. Sterile sample bottles were used for in pyrogallic acid-amended soil after incubation for
collection of samples for microbiological and 3 hours at 30 oC. The result was expressed as
enzyme analysis. Clean 250m amber reagent mg-1 dry soil 3h-1. Acid phosphatase activities were
bottles soaked in acid alcohol for 24 hours and determined as described by Tabatabai (14) using
rinsed thrice in deionized water were used for 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- The pH which ranged between 6.21 to 7.62 and
nitrohenol. moisture content followed the same pattern with
temperature. Soil organic matter, SO4, PO4 and
Microbiological analysis NO3 followed a gradient with lowest values at the
The bacteira diversity and population in each of range centre and gradually increased till the edge.
the sample soil were determined using various The 5-6m outside the range sample value was quite
culture media. The bacterial groups estimated were close to the control. Organic matter content ranged
the Total Aerobic heterotrophic Bacteria (TAHB), between 4.22-8.37 mg/g, NO3 was 0.52 - 3.91 mg/
the Nitrifying Bacteria (NB), and the Phosphate g while SO4 and PO4 had 2.12-4.82 mg/g and 16.37
Solubilizing Bacteria (PSB). The TAHB was - 29.78 mg/g ranges respectively (Table 1).In each
determined using Tryptone Soy Agar, NB with case, highest values were in the control while the
Mineral Salt Agar and PSB with Phosphate least were at the range centre soil samples.
Solubilzing Bacteria Medium as outlined in US The heavy metal concentrations increased with
Patent 6638730 (15). The soil samples were distance to the centre of the range. All the heavy
inoculated on te various media using the spread metals had their highest concentrations in the range
plate technique. centre soil sample, followed by the midway soil
sample before the edge sample. The values of soil
Analysis of data samples collected from 5-6m outside the edge
The results obtained in this study were subjected were close to those of the control. Zn had the
to statistical analysis using two main methods - highest concentration in the soil with values ranging
Correlation analysis for the enzymes activities with from 44.31 to 214.61 mg/g, followed by Pb (41.61-
heavy metal concentrations. This was also used 183.14 mg/g) and Cu, 41.54-172.31 mg/g. Cd was
for the bacterial loads and heavy metal the lowest (0.02-0.97 mg/g). The difference of
concentrations. The heavy metals and physico- the metal concentrations according to sampling
chemical parameters in soil sampling areas were area were statistically significant (P=0.05) (Table
compared using the standard deviation to test for
Table 3 shows the values of soil enzymatic
the differences.
activities which correlated negatively with the
heavy metals concentrations. Soil samples with high
RESULTS heavy metal concentrations showed low enzymatic
activities while those with low heavy metal
The results of the physicochemical parameters concentrations had higher soil enzymatic activities.
of the various soil samples are shown in Table 1. All the enzymes had significantly different activity
The soil temperature showed that values at the values at each sampling point.
Shooting range centre and midway were Among the soil enzymes, dehydrogenase had the
statistically different from the other areas sampled highest activities with a range of 0.79-8.24 mg g-
while that other areas were similar to each other. 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.

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

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
-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

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

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

Vol. 4, No. 2. 2008 Soil quality and enzyme activities

of these metals in the soil immediately out side the dehydrogenase and phosphatase activities. This
range, could be due to rain water runoff which observation could be early seen in this study. These
carry away some of the metals from the point of are enzymes produced by the microorganisms, it
pollution. Nwaugo et al. (10, 17) and Chinyere (9) therefore follows that the more the bacterial
had earlier reported similar observations as pollutant population, the higher the amount of enzymes
concentrations form a gradient away from the point released hence the higher the activity. The opposite
of pollution. applies true for low microbial density soil as
Observations of the physicochemical parameters reported in the shooting range centre.
suggest adverse effects on soil quality through the Observation of the various bacterial groups
shooting activities too. While the most affected shows that the most affected organisms were the
areas were slightly acidic, soil nutrients such as nitrifying bacteria. Nwaugo et al. (22) and Prescott
PO4, NO3 and SO4 including organic matter were et al. (19) stated that nitrifying bacteria are very
equally low in these highly contaminated areas. sensitive to environmental disturbances. Nwaugo
These suggest antimineralization and anti humifi- et al. (10) agree with Lee et al. (1) that high
cation effects on the soil which are unlike the situ- nitrogen compound in the soil will suppress soil
ation in the control sample and samples of soil out- nitrification process. This could be the case, as
side the shooting range. These observations agree nitro aromatics form the major components of
well with those of Lee et al. (18) in South Korea. military chemicals such as TNT used in Shooting
Lee et al. (1) also reported that high concentra- Ranges. PSB were equally affected adversely
tions of heavy metal reduce organic matter. These along with Aerobic heterotrophic bacteria. The
observations agree with Prescott et al. (19) and AHB are the sum total of all culturable aerobic
Pelczar et al. (5). This study therefore agrees that bacteria in the soil, hence they include even the
high heavy metal concentrations reduce soil nutri- phosphate solubilzing bacteria and some nitrifying
ent levels, soil moisture and pH. bacteria capable of growing on the Tryptone Soy
The results obtained on the enzymatic activities Agar used. The bioloads of these organisms
confirmed earlier findings that heavy metals affect positively correlated with the enzymatic activities,
soil enzymes adversely (3, 4, 8, 20). In all the as the higher the bioload, the higher the enzymatic
enzymatic activities estimated, the results activities.
correlated positively with the bioloads of microbial The results obtained in this study therefore
groups estimated as reported elsewhere. Heavy suggest that the Shooting range soil lacks soil
metals tend to inhibit the activities of these enzymes nutrients following reduced microbial and
including other processes in the microorganisms enzymatic activities. This study then suggests that,
resulting in low enzymatic activities and microbial if the soil could be used for any other purpose, the
population in contaminated soil. heavy metal level most be reduced and the soil
The most affected enzyme was dehydrogenase. rehabilitated. The rehabilitation becomes very
The activity of the enzyme at the shooting range essential, if the soil will be put into agricutlrual uses
centre was only 9.58% of the control value. The because most of the plants will bio accumulate
middle had only 14.56 % of the control value too. these metals in their tissues.
Similarly the urease value at the shooting range
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