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Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.

ASSESSMENT OF SOIL HEALTH IN DIFFERENT SITES USING


MICROBIOLOGICAL AND PHYSICOCHEMICAL PARAMETERS

1NWAUGO, V.O., 2EZENOBI, N., 2EKONG, U. AND 3OGBONNA, C.E


1Dept. Microbiology, Abia State University, Uturu Nigeria
2Dept. of Microbiology, University of Uyo, Uyo Nigeria
3Dept. of Environ. Resource Mgt, Abia State University, Uturu

ABSTRACT
Soil samples from different sites (residential, waste dump, market, vehicle maintenance
(mechanical village) and play ground were assessed for health status using physicochemical and
microbiological parameters in Okigwe, southern Nigerian. Conductivity (315-675um) and CEC
(28.3 49.3) were more prominent in waste dump site, followed by mechanic village but the
difference between residential and playground sites were not significant (P = 0.05). Microbial
and plant nutrients NO3, SO4, PO4 and organic matter were statistically higher in the dumpsite,
market, residential, playground and mechanic village in that order. Concentrations of Cu, Fe, Cd,
Cr, Zn, and Pb were significantly higher in the mechanic village soil samples, followed by the
dumpsite but fell below the toxic levels in the other sites analyzed. The most affected bacterial
group was the nitrifying bacteria, followed by the colifiroms, phosphate solubilizing bacteria and
the total heterotrophic bacteria, which were the highest. The lipolytic bacteria, though highest in
the dump site (4.3 x 104 cfulg) were higher in the mechanic village soil than the other sites. Soil
samples with high heavy metal concentrations had low bacteriological loads. All the enzymes
whose activities were analyzed, had their highest activities in the dump site, followed by the
market and least activities in the Mechanic village soil except Lipase which was highest only
second to dump site soil. The work therefore concludes that anthropogenic activities have
significant influence on soil health and that examination of soil health should be both
physicochemical and biological. Again, proper waste disposal system should be adopted by policy
makers to avoid environmental degradation.

Keywords: Different sites, organic matter, soil enzymes, soil health

INTRODUCTION

The soil is a complex mixture of several types of organic and inorganic materials including
various living biomass. It is also the primary resource on which all other terrestrial trophic
systems depend (Prescott et al., 2003, Pelczar et al., 2003). Generally, living systems are
important in the biogeochemical transformations taking place in soil including re-cycling of plant
nutrients, elements etc (Rich et al., 2003). The various organic and inorganic materials in the soil
determine the microbial spectrum that can be found in any given soil (Nwaugo et al., 2004,
Prescott, et al., 2003). This in turn, is influenced by the human activities in the particular site as
different activities cause the discharge of different substances into the environment (Humphries,
2007).
Man has put the soil or land into various uses depending on several factors. Such uses
range from residential, recreational to industrial and agricultural uses. Since these activities
introduce extraneous substances into the soil, the microbial biota, which is the major catalyst in
transformation taking place in soil, are affected. Ultimately, the fertility, structure and health of
soils are largely dependent on the functions and performance of the soil microbial community
(Humphries, 2007, Shi et al., 2002). The maintenance of soil quality to enable it perform its
Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.E

functions therefore include protection and management of the soil in such a way that the amount
of extraneous materials impacting on it is extensively regulated (Nwaugo et al., 2007b, Prescott et
al., 2003).
Assessment of soil health has recently shifted to include soil enzymes. This is to assess the
activities of both culturable and unculturable microbial entities involved in soil biogeochemical
transformations. This work therefore was designed to assess the health of soil samples from
different sites using not only the physicochemical properties but also microbiological indices as
indicators.

MATERIALS AND METHODS

The study area is Okigwe and it's surroundings. Okigwe lies in the typical rainforest region
of Guinea Savanna zone of Nigeria. The tropical climatic features characterize it. It is a semi-
urban human settlement where several activities take place as people of several professions and
trades are found there.

Sampling Sites and Method


Soil samples from five different areas were analyzed in this work. These were residential,
market and playground. Others were dumpsite and vehicle maintenance area (mechanic village).
Soil samples from the topmost 1-15cm of each site were aseptically collected. The samples
were sieved to remove particles of more than 2mm and then homogenized properly.
Physicochemical characterization was done within one-two weeks of collection, while
microbiological and enzymatic activities were evaluated within 1-2 days of collection.

ANALYSIS OF SOIL SAMPLES

Physicochemical Parameters
The soil temperature, pH and conductivity were assessed using the JENWAY
multipurpose tester HANNA 1910. The organic matter content was determined using the loss of
ignition method as described by Lee et al., (2002). This involved the use of furnace -MAC 2000.
The soil nitrate, (NO3), Phosphate (PO4), and cation exchange capacity (CEC) were determined
as described in AOAC (2005).
Soil samples for heavy metal determination were digested and analyzed using the
methods described in UNEP (2004) manual. The metals assessed were lead (Pb), cadmium (Cd),
Zinc (Zn), Copper (Cu), and Iron (Fe), which were determined using Atomic absorption
spectrophotometer (AAS).

Microbiological flora

The bacterial diversity and community structure were estimated in groups using specific
functions as parameters. Such bacterial groups include the nitrifying bacteria (NB), coliform
bacteria (CB), total heterotrophic bacteria (THB) and lipolytic bacteria (LB). These groups of
bacteria were evaluated using selective culture media. Nitrifying bacteria were determined using
modified mineral salt Agar, coliforms with McConkey Agar, Total heterotrophic bacteria with
Tryptone Soy Agar and Tributyrin Agar for lipolytic bacteria as described by Chessbrough,
(2001). The phosphate solubilizing bacteria were estimated using the method stated in US Patent
(2003). The soil samples were inoculated on the prepared culture media after ten-fold serial
dilution using sterile saline as diluent.
Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.E

Enzymatic analysis

The enzymes whose activities were analyzed in this work were dehydrogenase, lipase,
urease and phosphatases (acid and alkaline). The dehydrogenase was determined as described by
Cassida et al., (1964) using triphenyl tetrazolium chloride (TTC) as substrate. The TTC amended
soil was incubated at 300C for 6 hours. The triphenyl formazon formed was extracted with
methanol and measured at 485nM absorbance with the result expressed as TPF mg -1 dry soil 6h-
1.
The activities of both acid and alkaline phosphatases were determined according to
Tabatabai (1997) which involved the use of p-ntriophenyl phosphate under acidic (HCl 5.5) and
alkaline (NaOH 11) conditions. Results were read at 410nM and expressed as umol-p-
nitrophenol, which was released in the reaction.
The lipase activity was estimated as described by Sairubarumiyan et al., (2004) involving
the copper soap colorimetric method.

RESULTS

The prevalence of each bacterial group as observed in this work is shown in Table 1 as
indicated by the bacteriological loads. The most prevalent group was the Total heterotrophic
bacteria (THB) with a range of 3.7 x 106 5.7 x 107 cfu/g. Lowest values were observed at the
vehicle maintenance soil (mechanic village) while the highest was at the dumpsite. The nitrifying
bacteria were the least in all the sites (soil samples) analyzed with a range of 2.1 x 103 3.7 x 104
cfu/g. The range of bioloads were 2.3 x 103 4.3 x 104 and 3.1 x 104 6.1 x 105 cuf/g for lipolytic
and coliform bacterial groups respectively (Table 1).
The soil enzymes whose activities were assessed in this work showed values of various
ranges. While dehydrogenase had the highest activities in each soil sample with a range of 18.41
39.36 mg g-16h-1, the other enzymes had ranges that varied according to the site analyzed. In the
residential area soil, the least enzyme was lipase (2.13 mg g-h.1) just like the 2.07 mg g-3h-1,
observed in the playground soil sample was the least there too.
However, at the mechanic village area, the least enzyme was alkaline phosphatase
(2.07mg-1g-3h-1) while lipase was the least in the market place soil (2.92 mg g-3h). In all the soil
samples examined, the values for alkaline and acid phosphatase were not significantly different
from each other (P = 0.05), but dehydrogenase activities were statistically higher than all the other
enzymes analyzed. In lipase activities, the values obtained at the mechanic village soil were
significantly higher compared to other areas and enzymes as indicated by the values of ANOVA,
using enzymes and soil type as variables. Correlation analysis of the enzyme, dehydrogenase and
bioload of the THB positively correlated as the higher the bioload, the more the enzyme activities.
This same trend was also observed using lipolytic bacterial count and lipase activities. In the same
vein, the phosphatases showed the same trend with the phosphate solubilizing bacterial counts.
Table 4 shows the physicochemical parameters including the heavy metals whose
concentrations in the soil were analyzed. The value of the temperature observed ranged from
28.20C (play ground) to 30.60C but was not significantly different (P = 0.05). The pH also
showed slight variation with highest alkaline (8.2) at the dumpsite but slightly acidic (6.1) at the
mechanic village. Other values are between these two (6.1- 8.2).
The conductivity, PO4 and NO3 followed the same trend with highest at the dumpsite and
lowest at the mechanic village. The values obtained in the residential area soil and the playground
were not statistically different. (P = 0.05). However CEC, which had a range of 26.3 49.7, was
highest at the mechanic village but lowest at the playground (Table 4).
In the heavy metal concentration analysis, all of them had their highest concentrations in
the mechanic village soil samples, followed by the dumpsite while the results of the playground
Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.E

and residential soil were not significantly different (P = 0.05). While Zn had the highest values
(41.81 61.31 mg/l), the range for Cu and Cr were relatively very low with ranges of 0.032- 0.102
mg/l and 0.016-0.093mg/l and 0.016-0.093 respectively. Fe had concentration values of 10.27
(play ground) 36.79 mg/l (mechanic village). The values for the Cr were from 0.041- 0.102 mg/l .

DISCUSSION

Most of the physicochemical parameters analyzed in this work had their highest values in
the dump site except the CEC which was highest in the mechanic village soil. pH and
temperature were highest at the dumpsite which could be suggestive of the biogeochemical
transformations taking place. Olalade et al., (2007) reported a similar situation and stated that
most of the wastes deposited were biodegradable which could cause exothermic reactions and the
release of NH3 from the proteinous wastes. The NH3 dissolved in the soil moisture to cause a pH
rise (Nwaugo, et al., 2007b). Some of the wastes are organic substances containing NO3, PO4 and
other organic matters, which could increase the concentrations of such substance in the soil.
Some domestic animals like the dogs and rodents go to such waste dumps to scavenge for food.
These animals defecate there adding further organic and plant nutrients (Olalade et al., 2007,
Perham et al., 2002).
On the other hand, good sanitary conditions are often maintained in the case of residential
and playground areas while market soil may not necessarily receive as much wastes as dump sites.
This could explain the low values of the physicochemical parameters in such soil samples.
The mechanic village soil, though with high metal concentration, had lower conductivity
and CEC than the dumpsite. This suggests that the total concentration of metals in the soil may
not actually influence the bio availability of such metals which agrees with Nwaugo et al., (2007b)
and Canovas et al., (2003). The high particulate metallic substances and low moisture did not
allow for much ionization, hence the low conductivity and CEC. The reverse was the case in
dumpsite soil with high moisture content and release of metallic ions, which agrees with Olalade
et al., (2007). The market, residential and playground soil with low metallic wastes showed low
conductivity and CEC, which agrees with Humphries (2007), and Diaz-Ravina and Baath (1996).
Observations showed that all the metals assessed had their highest concentrations in
mechanic village soil (Fe, Pb, Zn, Cu, Cd and Cr followed by the dumpsite soil. This too, could be
expected. Vehicle parts contain alloys of most of these metals. Copper wires, Fe and Zn pans are
often used in construction and reconstruction of damaged vehicle bodies. The other metallic
counts could be contaminants or alloys of major ones. On the other hand Olalade et al., (2007)
reported that various substances including empty metallic cans, metallic rods and other discarded
metallic materials are common features of dumpsites in Nigeria, as there is no source separation
of wastes. The other sites examined in this work are not very much associated with metallic
substances. However, that values from residential areas are higher than playground could be due
to construction works at homes.
Microbiological analysis in this work agrees with Nwaugo et al., (2007c,d), Chander et al.,
(2000) and Kupperman and Carreiro (1997). The bioloads of most of the bacterial groups analysed
followed the soil nutrients pattern and metallic concentrations. The waste dumpsite, with its high
nutrients content - (PO4, NO3, organic matter) had the highest bacterial loads. These substances
served as nutrients for bacterial survival and proliferation. On the other hand, the harsh conditions
of the mechanic village soil could not allow much bacterial proliferation. The low level of
bacterial load in the playground and residential areas could be suggestive of low nutrient level and
poor environmental management.
The least bacterial group was the nitrifying bacteria while the highest was the total
heterotrophic one. The THB is a summation of all viable bacteria, while the other ones are
Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.E

fractions of this THB. Nwaugo et al., (2007c) and Prescott et al., (2003) had reported that
nitrifying bacterial are easily destabilized in soil pollution which agrees with this work.
Observations equally show that the lipolytic bacterial count in the mechanic village was relatively
high compared to other bacterial groups and sites. This slight shift in bacterial load could be due
to substrate induction. Various oily wastes contaminate the mechanic village soil, which in turn
serve as substrates for the lipolytic bacterial. Nwaugo et al., (2006) and Li et al., (2006) had
earlier observed similar situation. Results obtained also showed the dumpsite as having the
highest coliform and phosphate solibilizing bacterial counts with mechanic village having the
least. Nutrient availability and environmental conditions could easily be suggestive of this
situation too.
Results obtained from the enzymatic analysis followed the same pattern with bacterial
loads and nutrient availability. In addition heavy metal concentrations also played a significant
role as soil with high concentrations had low enzymatic activities except lipase enzyme. The
situation with lipase activities agrees with the substrate availability and bacterial load. Generally,
dehydrogenase had the highest activity while lipase had the lowest. Acid and alkaline
phosphatases did not show much variation. All microorganisms produce dehydrogenase while the
other three enzymes were specific or substrate induced. This then suggests that all bacterial
species that produce lipase and phosphatase also produce dehydrogenase while the reverse is not
the case with dehydrogenase producers. This work however agrees with Lee et al., (2002) and
Oliveira and Pampulha (2006) that soil enzyme activities are closely related to the prevalence of
microorganisms.
The enzymatic activities values obtained in this work seem to agree with those of other
authors. Such ranges in enzymatic activities have been reported by Li et al., (2005), Lee et al.,
(2002) and Nwaugo et al, (2007c) in their various work. This then shows that except in extreme
cases, the activity ranges of soil enzymes are similar in most impacted soil of similar climatic
zones.
In conclusion, microbial structure and functions in any soil are influenced by activities on
it and the use to which it is put The work also suggests that the assessment of soil health should
be enlarged to include all necessary areas while soil management should be done to ensure good
quality soil everywhere. This study also calls for proper control of wastes disposal in properly
designated areas no matter where they are generated to avoid environmental degradation.
Journal of the Environment, Vol. 2(1): 81-89, 2008. ISSN:0795-0977 Nwaugo, V.O., Ezenobi N. Ekong & Ogbonna, C.E

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