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INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCES Volume 3, No 1, 2012
© Copyright by the authors - Licensee IPA- Under Creative Commons license 3.0
Research article ISSN 0976 – 4402
Received on June, 2012 Published on July 2012 570
Bacteriological analysis of water and soil samples around Nanjangud
industrial area, Mysore district, Karnataka, India Rajanna A.H, Shyamala D.C, Belagali S.L
DOS in Environmental Science, University of Mysore, Manasagangothri, Mysore-6,
Karnataka, India.
doi:10.6088/ijes.2012030131056
ABSTRACT
The present study was undertaken to evaluate bacteriological pollution level of Kabini river
water, industrial effluents and soil samples around Nanjangud industrial area, situated near
Mysore in the state of Karnataka during 2009. 33 water samples and 13 soil samples were
collected and subjected for bacteriological examination. Standard plate count and most
probable number tests were performed by presumptive, confirmative and completed tests
using respective agars. Further, morphological and biochemical characteristics of the
microbial isolates were carried out and bacterial populations were identified as Escherichia
coli, Klebsiella sp., Enterobacter aerogenes, Bacillus sp., Salmonella sp. and Pseudomonas sp.
Similarly, fungi like Aspergillus niger, Yeast, Rizopus, Penicillium and Curvelarea were
identified and Aspergillus flavous and Fussarium sp., were more abundant in soil samples.
The MPN in the water samples were observed upto 150 MPN/100ml, which signified
undesirable level of water pollution in the area covered and was not suitable for drinking
purpose without adequate treatment and appropriate programmes must applied to educate the
general populace on the need to purify water to make it fit for drinking and other domestic
purposes.
Keywords: Nanjangud indusial area, Kabini river, bacteria, fungi.
1. Introduction
Water plays a vital role in the proper functioning of the earth ecosystem (Ajayi and Adejumo
2011). The quality of water for drinking deteriorates due to inadequacy of treatment plants,
direct discharge of untreated sewage into rivers (UNEP 2001). Rivers are being polluted by
indiscriminate disposal of sewerage, industrial wastes and plethora of human activities, which
affect their physico-chemical and biological quality (Koshy 1999). Coliforms are the major
microbial indicators of monitoring water quality (Brenner et al., 1993). Human faecal
material generally caused greater risk to human health because it contains human enteric
pathogens (Scott et al., 2003). Faecal polluted water may cause health hazard for consumers
due to the presence of several microbial pathogens, including bacteria, viruses, fungi, and
protozoa (Mohamed et al., 2008). The soil micro biota is instrumental in the degradation and
synthesis of organic compounds (Perkinson and Coleman 1991). Urbanization and
industrialization has badly affected quality of water and soil. Consequently, continuous
monitoring of water quality is necessary, particularly in the industrial areas (Will and Yeh
1987). In fact, attention has been paid for monitoring and assessment in microbiological
quality of water resources all over the world (Lindskog and Lindskog, 1998; Fernandez -
Alvars et al., 1991; Khalafalla et al., 1993). A powerful monitoring program is needed to
provide reliable information about the current water quality. Therefore, the present study was
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 571
conducted to assess the microbial quality of water and soil samples around Nanjangud
industrial area, situated near Mysore in the state of Karnataka.
1.1 Study area
Figure 1: Image showing the map of the sampling stations of the study area
Nanjangud is spread over from 12º 06′ 39″
N longitude to 76
º 33′
46″ E latitude and 12
º 07′
56″
N longitude to 76º 42′
9″
E latitude. The total geographical area of Nanjangud is 991 Sq, km.
According to 2001 census the total population of the taluk was 3, 60, 223. Nanjungud is rated
as the fastest growing industrial area. It is a home to many of industries, which are spread in
532 acres, across Kabini river. There are 36 major industries and several medium and small
scale industries which are located on the banks of Kabini river, a major tributary of Cauvery
river. Some of the mainly located industries in Nanjungud are Jubilant Organosys Ltd.
(Pharmaceutical), AT & S India Pvt. Ltd. (Electronic), Nestle India Ltd. (Food), Eastern Silk
industries Ltd. (Textile), Zenith (Textile) etc. Most of the people are dependent on the Kabini
river for their daily domestic activities in the study area. Sampling stations of the study area
under investigation is depicted in Table-1.
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 572
Table 1: Table showing the fresh water and waste water sampling stations of the study
area
Sampling stations of fresh water samples
S-1 Channel water (Kallahalli)
S-2 Kabini river water (Kallahalli)
S-3 Kabini river water (Old bridge)
S-4 Kabini river water (Kathwadipura)
S-5 Channel water (Kathwadipura)
S-6 Kabini river water (Guest house)
S-7 Ground water (Korihundi)
S-8 Ground water (Kallahalli)
S-9 Kabini river water (Kathwadipura) (100 meters away from S-4 sample)
S-10 Kabini river water (Kathwadipura) (one kilometer away from AT & S. Pvt.Ltd.)
S-11 Kabini river water (Kathwadipura) (neare to AT & S India Pvt. Ltd.)
S-12 Kabini river water (Kathwadipura) (100 meters away from S-11 sample)
S-13 Ground water (Kathwadipura Village entrance)
S-14 Ground water (Kathwadipura Village interior)
S-15 Kabini river water (Kathwadipura) (100 meters away from S-12 sample)
S-16 Kabini river water (Kathwadipura) (100 meters away from S-15 sample)
S-17 Kabini river water (Kathwadipura) (half a kilometer away from the industrial
area)
S-18 Kabini river water (Kathwadipura)
S-19 Kabini river water (Kathwadipura) (neare to Nestle India Pvt. Ltd.)
S-20 Kabini river water (Kathwadipura) (100 meters away from S-19 sample)
S-21 Kabini river water (New bridge)
S-22 Kabini river water (Kathwadipura temple)
S-23 Rampura channel water (near to South India paper mills
S-24 Rampura channel water (100 meters away from S-23 sample
S-25 Inside the Nestle India Ltd. (KIADB water)
S-26 Inside the Brakes India Ltd. (Ground water)
S-27 Inside the Rubin Dairy Industry (Ground water)
S-28 Inside the Jubilant Organosys Ltd. (KIADB water)
S-29 Inside the Zenith Textile industry (KIADB water)
Sampling stations of waste water samples
ES-1 Inside the Nestle India Ltd. Industry
ES-2 Inside the Brakes India Ltd. Industry
ES-3 Inside the Rubin Dairy Industry
ES-4 Inside the Zenith Textiles Industry
Table 2: Table showing the soil sampling stations of the study area
Sampling stations for soil samples
SL-1 River bank (Kallahalli)
SL-2 Channel side (Kallahalli)
SL-3 Outside the Jubilant Organosys Ltd. industry
SL-4 Outside the Rubin Dairy industry
SL-5 Outside the Ayurvedic industry
SL-6 Inside the Nestle India Ltd. industry (North side)
SL-7 Inside the Nestle India Ltd. industry (East side)
SL-8 Inside the Brakes India Ltd. industry (ETP side)
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 573
SL-9 Inside the Brakes India Ltd. industry (Storm water channel side)
SL-10 Inside the Brakes India Ltd. industry (Nursery side)
SL-11 Inside the Rubin Dairy industry
SL-12 Inside the Jubilant Organosys industry
SL-13 Inside the Zenith textiles industry (canteen side)
2. Materials and method
The water samples were collected in sterilized polythene bottles and the soil samples were
collected in sterilized polythene bags for the bacteriological analysis. Most probable number
tests were performed by using multiple tube fermentation technique, following presumptive
coliform test, confirmed test and completed test using agar media which includes Nutrient
agar, Lactose broth, Eosin Mythylene blue etc., and for the examination of microbes in soil,
1g of soil was suspended into 10 ml of sterile saline solution and serial diluted further upto
10-4
dilution aseptically. 0.1 ml of diluted soil suspension was poured onto nutrient agar plate
using spread plate technique. Plate were incubated for 24 hours at 37oC and bacterial colonies
are isolated and further pure culture were subjected to various morphological and
biochemical characterization tests like, Urease, Hydrogen sulphide, Starch hydrolysis,
Gelatin hydrolysis, Catalase, Oxidase, Indole, Methyl-red Voge-Proskauer, Citrate utilization,
and Nitrate utilization to determine the identity of the bacteria isolates and were carried out
according to the procedure given in Practical Microbiology by R.C.Dubey and
D.K.Maheshwari (2002).
3. Results and discussion
The experimental results of the present study are summarized in the Tables 3, 4, 5 and 6.
Biochemical tests for Bacteria in water samples are represented in Figure 2, identified
bacteria in soil samples are given in figure.3 and Identified fungi in soil samples are shown in
figure. 4.
Table 3: Table showing bacterial observation in the water samples
Sample No. Total plate count MPN/100 Organisms
identified
S-1 247 28
Escherichia
coli
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-2 274 28
Escherichia
coli
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-3 204 28
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 574
Flavobacterium
sp.
S-4 350 150
Escherichia
coli
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-5 303 28
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
Flavobacterium
sp.
S-6 221 150
Escherichia
coli
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-7 16 - Bacillus sp.
S-8 15 3 Bacillus sp.
S-9 415 39
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-10 180 21
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-11 328 21
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-12 221 11
Klebsiella sp.
Enterobacter
aerogenes
Bacillus sp.
S-13 30 3
Bacillus sp.
Enterobacter
aerogenes
S-14 20 - Enterobacter
aerogenes
S-15 180 28
Klebsiella sp.
Salmonella sp.
Pseudomonas
sp.
S-16 192 150 Klebsiella sp.
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 575
Escherichia
coli
Pseudomonas
sp.
S-17 172 11
Klebsiella sp.
Salmonella sp.
Pseudomonas
sp.
S-18 124 28
Klebsiella sp.
Escherichia
coli
Salmonella sp.
Proteus sp.
S-19 312 11
Klebsiella sp.
Escherichia
coli
Pseudomonas
sp.
Salmonella sp.
Bacillus sp.
S-20 609 11
Klebsiella sp.
Bacillus sp.
Pseudomonas
sp.
Salmonella sp.
Escherichia
coli
S-21 271 43
Klebsiella sp.
Bacillus sp.
Pseudomonas
sp.
Escherichia
coli
S-22 473 120
Pseudomonas
sp.
Escherichia
coli
Proteus sp.
Proteus
vulgaris
Most proteus
mirabilis
S-23 122 120
Klebsiella sp.
Proteus sp.
Pseudomonas
sp.
Salmonella sp.
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 576
Escherichia
coli
S-24 275 120
Pseudomonas
sp.
Escherichia
coli
Proteus sp.
S-25 56 28
Klebsiella
pneumoniae
Pseudomonas
aerugionosa
S-26 3 - Bacillus sp.
S-27 6 - -
S-28 65 20 Klebsiella sp.
Salmonella sp.
S-29 74 150
Klebsiella sp.
Pseudomonas
aerugionosa
Escherichia
coli
Proteus sp.
Table 4: Table showing biochemical characteristics of isolates
Test W1 W2 W3 W4 W5 W6 W7 W8
Catalase + + + + + + - -
Oxidase - - - + - - - -
Indole + - - + + - - -
Methyl-red + - - - + + (+) -
Voge-Proskauer - + + + - + + +
Citrate utilization - + - + - - + +
Urease - + - - + - + -
Hydrogen
sulphide
ND - + + - - - +
Starch hydrolysis - + - - - - - -
Gelatin hydrolysis - (+) + + + + (+) +
Nitrate utilization + + + + + (+) + -
W1 = Escherichia coli W2 = Klebsiella sp.
W3 = Bacillus sp. W4 = Salmonella sp.
W5 = Proteus sp. W6 = Flavobacterium sp.
W7 = Enterobacter aerogenes W8 = Pseudomonas aerugionosa
ND = Not- detected.
Microbial investigation of the studied water samples show that, the value of total plate count
and coliforms were high in river water and less in ground water. The high total plate counts
observed in river water indicated the presence of high organic matters and related nutrient
sources. The most probable number (MPN) for the presumptive total coliform count of the
river water samples ranged from 11 to 150 MPN/100 ml and zero to 3 in ground water
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 577
samples, it might be due to proximity to toilet facilities and various human activities around
borewells. More number of bacterial populations in most of the present examined samples
indicates that, the runoff of untreated water is grossly contaminated. The coliform count on
EMB agar plate also showed more number of coliforms in S-4, S-6, S-16, S-21, S-22, S-23,
S-24 and S-29 samples and S-1, S-2, S-3, S-5, S-9, S-10, S-11, S-12, S-15, S-17, S-18, S-19,
S-20, S-25 and S-28 samples, indicating medium number of coliforms. Ground water
samples like S-7, S-8, S-13, S-14, S-26 and S-27 had very negligible coliforms. The results
reflect poor hygienic practices by surrounding villagers and the possible sources of
contamination by both man and other animals. Most of the people discharge their waste into
river directly rather than poor people living near the river discharge their shit at river bank
(Srivastava et al 2011). Total plate count is considered to represent the contamination from
the natural environment and also indicate that the presence of bushes and shrubs makes likely
possible that, smaller mammals may have been coming around these water bodies to drink
water, thereby passing out faeces into water. The complete coliforms test showed a positive
test for all the water samples. The biochemical characteristics of the isolates obtained from
these water samples are shown in Table 4. The identified isolates include Salmonella species,
Escherchia coli, Pseudomonas aerugionosa, Enterobacter aerogenes, Bacillus sp., Proteus
sp., Klebsiella sp., Flavobacterium sp., and Acinetobacter sp. Results show an indication of
contaminant sources from the fact that, people living on nearby bank of river directly dispose
off their domestic sewages, human and animal excreta etc.
Figure 2: Images showing the biochemical tests for bacteria in water samples
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 578
Table 5: Table showing bacterial populations of soil samples
Sample No. Identified soil bacteria Total viable count
S-1
Enterobacter aerginosa
Pseudomonas aerugionosa
Coryne-bacterium xerosis
65
S-2 Enterobacter aerginosa
Pseudomonas aerugionosa Salmonella sp. 51
S-3
Klebsiella sp.
Pseudomonas sp.
Bacillus sp.
49
S-4
Enterobacter aerginosa
Klebsiella sp.
Bacillus sp.
15
S-5
Enterobacter aerginosa
Klebsiella pneumoniae
Salmonella sp.
14
S-6
Proteus sp.
Enterobacter aerginosa
Klebsiella sp.
Pseudomonas sp.
Bacillus sp.
150
S-7
Enterobacter aerginosa
Acinetobacter sp.
Bacillus sp.
Klebsiella pneumoniae
Pseudomonas aerugionosa
Proteus sp.
159
S-8
Klebsiella pneumoniae
Enterobacter aerginosa
Pseudomonas aerugionosa
Proteus sp.
Bacillus sp.
200
S-9
Proteus sp.
Klebsiella sp.
Pseudomonas sp.
Bacillus sp.
Acinetobacter sp.
235
S-10
Proteus sp.
Klebsiella sp.
Pseudomonas sp.
Acinetobacter sp.
300
S-11
Klebsiella pneumoniae
Pseudomonas aerugionosa
Acinetobacter sp.
Bacillus sp.
200
S-12
Klebsiella sp.
Pseudomonas sp.
Bacillus sp.
145
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 579
Acinetobacter sp.
S-13
Salmonella sp.
Bacillus sp.
Acinetobacter sp.
123
Figure 3: Images showing the identified bacteria in soil samples
Various groups of microorganisms were isolated and identified during the study in soil
samples they include Proteus sp., Klebsiella sp., Pseudomonas sp., Bacillus sp.,
Acinetobacter sp., and Enterobacter aerginosa. These are natural micro flora of soil.
Particularly dangerous proved to be bacteria of salmonella genus. Although Salmonella
presence confirmed in the soils examined was occasional, still even their small count should
be alarming because they can easily spread under favourable conditions and make a serious
source of environmental pollution (Tymczyna 2000). The presence of Enterobacter
aerginosa sp. indicates the contamination of soil by the faecal matter. Actual habitat of
Enterobacter is animal intestine but their presence in soil indicates contamination by faeces
of animals or human beings. These Enterobacter are pathogens and harmful to human beings,
if injested.
Table 6: Table showing fungal colonies in soil samples
Sample No. Identified fungi
No. of colonies
Aspergillus flavous 5
Aspergillus ficum 10 SL-1
Curvelarea 2
Aspergillus niger 39 SL-2
Fusarium 40
Aspergillus niger 12
Aspergillus flavous 7 SL-3
Fusarium 25
Fusarium 20
Aspergillus flavous 12 SL-4
Aspergillus niger 8
SL-5 Aspergillus flavous 15
Fusarium 20
Aspergillus flavous 12
Aspergillus ficum 5
SL-6
Curvelaria 12
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 580
Fusarium 10
Fusarium 11
Aspergillus flavous 9
SL-7 Curvelarea 2
Rhizopus 1 SL-8
Fusarium 2
Aspergillus flavous 5 SL-9
Fusarium 10
Aspergillus flavous 10
Fusarium 17 SL-10
Yeast 5
Aspergillus flavous 12
Yeast 7 SL-11
Fusarium 25
Penicillium 3
Fusarium 5
Aspergillus flavous 4 SL-12
Rizopus 8
Aspergillus flavous 9 SL-13
Aspergillus niger 10
Identified fungi of soil samples under investigation illustrated in table-6 mainly, Aspergillus
niger, Yeast, Aspergillus flavous, Rizopus, Penicililum and Curvelarea. Aspergillus flavous
and Fussarium sp. are more abundent in all soil samples of the study area. Aspergillus sp. are
mainly plant and animal pathogens which increase due to food contamination and majority
are toxic to soil. Fusarium sp. is commonly found in soil and majority of them are plant
pathogens and are from food origin. The Rizopus rapidly grows white colored fungus swarms
over entire plate. These species are commonly seen in laboratory contamination. Penicillium
mature cultures are usually greenish blue in color. These fungi are using antibiotic condition
producing citrus fruit contaminants and soil inhabitant. Yeast is present in cheese and food
contaminations, some of the species are used in food industry and fermentation process. The
above results show that, most of the fungi are harmful to plants and animals.
Figure 4: Images showing identified soil fungi of the study area
4. Conclusion
The results of the microbiological analysis of water around Nanjangud industrial area have
shown that, most of the samples of river water have higher total coliforms, because of
Bacteriological analysis of water and soil samples around Nanjangud industrial area, Mysore district,
Karnataka, India
Rajanna .A.H, Shyamala .D.C, Belagali.S.L
International Journal of Environmental Sciences Volume 3 No.1, 2012 581
contaminant sources from the fact that, people living in the river areas directly dispose off
their domestic sewage, human and animal excreta and food industrial effluents are also one
of the source of environmental pollution through the discharge of the effluents into the
water body and on soil system. From the bacteriological points-of-view, the water sample
was not suitable for drinking purpose without adequate treatment and it is suggested that,
there should be proper treatment of industrial effluents to minimize the pollutants load in
water quality of River Kabini and some immediate measures have to be taken to maintain
water and soil quality around Nanjangud industrial area.
Acknowledgement
Mr.Rajanna A.H and Mrs. Shyamala D.C are grateful to Ministry of Environment and Forests
(MOEF), New Delhi for providing financial assistance to carry out the above work.
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