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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.

[email protected]

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.


Karnataka, India



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-13 Ground water (Kathwadipura Village entrance)

S-14 Ground water (Kathwadipura Village interior)



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-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)


Karnataka, India



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.


Karnataka, India



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.


Karnataka, India



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.


Karnataka, India



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


Karnataka, India



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


Karnataka, India



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


Klebsiella sp.

Bacillus sp.

15

S-5


Klebsiella pneumoniae

Salmonella sp.

14

S-6

Proteus sp.


Klebsiella sp.

Pseudomonas sp.

Bacillus sp.

150

S-7


Acinetobacter sp.

Bacillus sp.



Proteus sp.

159

S-8




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



Acinetobacter sp.

Bacillus sp.

200

S-12

Klebsiella sp.

Pseudomonas sp.

Bacillus sp.

145


Karnataka, India



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 niger 8

SL-5 Aspergillus flavous 15

Fusarium 20


Aspergillus ficum 5

SL-6

Curvelaria 12


Karnataka, India



Fusarium 10

Fusarium 11


SL-7 Curvelarea 2

Rhizopus 1 SL-8

Fusarium 2


Fusarium 10


Fusarium 17 SL-10

Yeast 5


Yeast 7 SL-11

Fusarium 25

Penicillium 3

Fusarium 5


Rizopus 8


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


Karnataka, India



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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