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Sci (2015) 4(7): 577-584

ISSN: 2319-7706 Volume 4 Number 7 (2015) pp. 577-584

Original Research Article

Bacteriological and Physico-Chemical Assessment of Municipal
Wastewater from Buldana District, India

A.M. Garode1 and N.A. Sonune2*

P.G. Department of Microbiology, Shri Shivaji Science College, Chikhli, Dist.
Buldana (M.S.), India
School of Life Sciences, SRTM University, Nanded (M.S.), India
*Corresponding author


The present study was conducted for bacteriological and physicochemical analysis
of municipal wastewater. The bacteria were isolated from municipal wastewater
Keywords samples as well as sludge samples. Total 44 bacteria were isolated on nutrient agar
medium. The isolated bacteria were belonging to genera of Enterobacter, Bacillus,
Municipal Paenibacillus, Klebsiella, Escherichia, Micrococcus, Salmonella, Pseudomonas,
wastewater, Proteus, Flavobacterium, Alcaligenes, Serratia, Aerobacter, Stenotrophomonas,
Isolation, Planococcus and Staphylococcus. The most of them were industrially important
Physico- enzyme producer whereas wastewater also contains pathogenic bacteria. The
chemical physicochemical analysis of four cities of Buldana district revealed that pH of
analysis, municipal wastewater ranging from 6.7 to 7.2. The BOD and COD values were
BOD, ranging from 72 mg/L to 238 mg/L and 115 mg/L to 480 mg/L respectively. The
COD nitrate values were ranging from 44.5 mg/L to 210 mg/L. The concentration of
phosphate was ranging from 0.98 mg/L to 2.1 mg/L whereas ammonical nitrogen
was ranging from 28.5 mg/L to 67 mg/L. These results reveals that pH and
phosphate were within the permissible limit whereas BOD, COD, ammonical
nitrogen and nitrate were above the permissible limits in accordance with World
Health Organization standards.


Domestic wastewater is the water that has The rapidly increasing human population
been used by a community and which and industrialization have placed
contains all the materials added to the water tremendous pressure on the natural water
during its use. It is composed of human resources and their quality (Sinha and
body wastes i.e. faeces and urine together Shrivastava, 1995). Because of such
with the water used for flushing toilets and problems, the river water usually receives
sullage. It is objectionable in appearance and untreated sewage, domestic waste, industrial
hazardous in content, mainly because of the and agricultural effluents that results in
number of disease-causing pathogenic pollution of several rivers in India. During
organisms (Mara, 2004). the last several decades the water quality of

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

the Indian River has been deteriorating due from Buldana district. Zip-lock plastic bag
to continuous discharge of industrial waste was used for sludge sample which was taken
and domestic sewage (Krishnan et al., 2007; from 4-5cm deep from the surface by the
Smitha et al., 2007). help of spatula. The wastewater samples
were collected in clean polythene bottle
WHO estimated that about 80% of water fitted with screw caps. Both zip-lock plastic
pollution in developing country, like India is bag and spatula were autoclaved before used
carried by domestic wastes. The improper and polythene bottle was washed with
management of water systems may cause distilled water. The wastewater samples
serious problems in availability and quality were collected for physicochemical analysis
of water. The water quality characteristics is from open drainage of four cities namely
denoted by knowing the physicochemical Buldana, Chikhli, Khamgaon and Mehkar of
parameters like pH, BOD, COD, Buldana district. The samples were collected
Ammonical nitrogen, nitrate, phosphate etc. from February 2012 to April 2012 according
(Subba Rao and Subba Rao, 1995). to standard procedures from American
Public Health Association (APHA, 1998).
In India, municipal and industrial waste is The samples collection time was from 8.00
not properly treated, especially excreta and to 10.0 am at morning.
other liquid waste from households and the
community which leads to a serious health Isolation and identification of bacterial
hazard and the spread of infectious diseases isolates: The sludge samples and
which ultimately leads to unhygienic wastewater samples were serially diluted
conditions and thereby to rise in the health and inoculated on the nutrient agar medium
problems. Kulkarni et al. (1996) reported by spread plate method. The plates were
out break enteric fever in village incubated at 370C for 24 h. The
Katkalamba, in Nanded district, Maharashtra morphologically different colonies were
in 1995. This was attributed to fecal isolated and purified and subculture on
contamination of water. Sathe et al. (1983) nutrient agar slants. These slants were kept
reported an explosive epidemic of typhoid at 40 C for preservation. Bacterial strains
fever occurred in Sangli Town in were examined for their colony and cell
Maharashtra State, India, between December morphology, motility, Gram and spore
1975 and February 1976. In 1955, an staining and standard biochemical tests such
outbreak of hepatitis in Delhi, India and in as Catalase, Oxidase, IMViC tests,
1817, major epidemic of cholera occurred in fermentation of carbohydrates, Nitration
Calcutta, India (U.S. Environmental reduction and H2S. Along with this, the
Protection Agency Washington, D.C. enzyme (amylase, protease and lipases)
PIPELINE Summer 7:3). The aim of producing activity was also checked. The
present study was microbiological and isolated bacteria were identified according
physicochemical analysis of municipal to Bergey s Manual of Determinative
wastewater. Bacteriology.

Materials and Methods Physicochemical analysis of wastewater:

The wastewater samples were collected
Sample collection and site: The sludge and from four cities such as Buldana, Chikhli,
wastewater samples were collected for Mehkar and Khamgaon from Buldana
isolation of bacterial from various places district. The samples were analyzed for

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

various physicochemical characters such as of these 23 bacterial isolates, 6 bacterial

pH, BOD, COD, ammonical nitrogen, isolates were amylase producer, 19 isolates
nitrate and phosphate. The experiments were were protease producer and 20 bacterial
performed in triplicate sets. The isolates were lipase producer. The 23
physicochemical analysis was done bacterial isolates were B. cereus (2),
according to APHA (1998). Paenibacillus borealis (1), Alcaligenes
faecalis (1), E. coli (1), Serratia ficaria (1),
Results and Discussion Serratia liquefaciens (1), Enterobacter
aerogenes (1), Bacillus sp. (1), Aerobacter
In present study, total 44 bacteria were hydrophilia (2), Pseudomonas aeruginosa
isolated on nutrient agar medium. Out of (1), Stenotrophomonas maltophilia (1),
these, 21 bacteria were isolated from Planococcus salinarum (1), Alcaligenes sp.
wastewater samples. Among these, 9 (1), Klebsiella sp. (1), Pseudomonas sp. (2),
bacterial isolates were Gram positive S. aureus (3), S. epidermidis (1) and Proteus
whereas 12 bacterial isolates were Gram sp. (1).
negative. All the 21 bacterial isolates were
catalase positive. Out of 21 bacterial Wan Ishak et al. (2011) isolated B.
isolates, 11 isolates were oxidase positive licheniformis from activated sludge and
whereas 10 isolates were oxidase negative. compost for municipal solid waste treatment
The total 10 bacterial isolates were spore system. Al-Zubeiry (2005) reported isolation
former while 11 were non spore former. All of Staphylococcus aureus, Pseudomonas
the isolates were tested for enzyme aeruginosa, Bacillus cereus, E. coli and
production such as amylase, protease and Salmonella sp. from raw sewage, secondary
lipases. Among these, 8 bacterial isolates effluent and dewatered sludge. Similarly
were amylase producer, 13 isolates were Garode and Sonune (2014) reported
protease producer and 11 bacterial isolates Paenibacillus borealis isolated from
were lipase producer. These 21 bacterial domestic wastewater. Many researchers
isolates were Enterobacter intermedius (1), (Mendes and Nascimento, 1991; Mitchel,
B. subtilis (2), B. cereus (1), B. licheniformis 1976; Ponamareva et al., 1994; Wilcox et
(2), Paenibacillus sp. (1), K. pneumoniae al., 1983) reported that the activated sludge
(2), E. coli (2), Enterobacter sp. (1), from municipal and industrial wastewater
Bacillus sp. (2), Micrococcus varians (1), contain great number of microorganisms
Salmonella sp. (1), Klebsiella sp. (1), and the genera found most frequently are
Pseudomonas sp. (1), Proteus sp. (2) and Pseudomonas, Bacillus, Achromobacter,
Flavobacterium sp. (1). Enterococcus, Acinetobacter, Aeromonas,
Alcaligenes, Arthrobacter, Escherichia,
In present study, total 23 bacteria were Salmonella, Proteus, Streptococcus,
isolated from sludge samples. Among these Staphylococcus, Micrococcus,
bacterial isolates 7 isolates were Gram Corynebacterium, Clostridium and
positive whereas 16 isolates were Gram Penicillium.
negative. All the isolated 23 bacteria were
catalase positive. Among 23 bacterial In present study most of the bacterial
isolates, 11 isolates were oxidase positive isolates showed enzyme producing ability
whereas 12 isolates were oxidase negative. such as amylase protease and lipases. Such
The total 8 bacterial isolates were spore enzyme producing bacterial isolates are
former while 15 were non spore former. Out industrially important as well as they may be

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

used for bioremediation of municipal Physicochemical analysis of wastewater

wastewater. In our study, B. subtilis, B. from Chikhli city:
cereus, B. licheniformis, Paenibacillus sp.
and Pseudomonas sp. etc were found to be The samples from Chikhli city was done at
enzyme producer. Garode and Sonune alternate days and total five samples were
(2013) also reported amylase producing collected and analyzed for their
Bacillus sp. and Paenibacillus sp. from physicochemical characters. The
domestic wastewater. Similarly Mobarak- physicochemical results of municipal
Qamsari et al. (2011) also reported lipase wastewater from Chikhli were showed in
producing Pseudomonas aeruginosa from table 2.
wastewater of an oil processing plant.
The pH of wastewater from Chikhli were
Physicochemical analysis of wastewater ranging from 6.80.01to 6.90.02. All the
from Buldana city: five values of pH were below the
permissible limit. The BOD values of all
Total five samples were collected from samples were above the permissible limit.
Buldana city. The sampling was done at The BOD was ranging from 785.88 mg/L
alternate days. The results from table1 to 13610.2 mg/L. The minimum value of
suggested that pH of wastewater from COD was 1159.55 mg/L whereas the
Buldana city was ranging from 6.80.01 to maximum value was 26116.34 mg/L. Out
7.00.12 and it was within the permissible of the five samples, one showed higher
limit. The lowest value of BOD was value the permissible limit and it was
769.96 mg/L whereas highest value was 26116.34 mg/L. The other four samples
14418 mg/L. All the BOD values were showed less value than permissible limit and
above the permissible limit. The COD they were 15624.52 mg/L, 21512.54
values were ranging from 13413.0 mg/L to mg/L, 22025.14 mg/L and 1159.55 mg/L.
26422.27 mg/L. Out of the five samples The ammonical nitrogen was ranging from
one sample showed high COD value than 346.8 mg/L to 541.2 mg/L. In these cases,
permissible limit and it was 26422.27 one value was above the permissible limit
mg/L. The other COD values were within i.e. 541.2 mg/L whereas remaining four
the permissible limit showed in table 1. The values were within the permissible limit and
ammonical nitrogen was ranging from they were 385.05 mg/L, 464.54 mg/L,
4614.18 mg/L to 6712.0 mg/L. Out of the 346.8 mg/L and 433.42 mg/L. The nitrate
five samples; two samples showed less was ranging from 44.55.5 mg/L to
concentration than permissible limits i.e. 14514.0 mg/L. The lowest value of nitrate
4614.18 mg/L and 484.0 mg/L. The was 44.55.5 mg/L and highest value was
remaining three samples showed higher 14514.0 mg/L. All the samples showed
concentration than permissible limit and higher values of nitrate than permissible
they were 6712.0 mg/L, 55.5 19.37 mg/L limit. The phosphate was below permissible
and 66.510.04 mg/L. The nitrate was limit and ranging from 1.010.08 mg/L to
ranging from 96.513.44 mg/L to 1665.0 2.10.14 mg/L.
mg/L. All the nitrate values were above the
permissible limit. The phosphate values of Physicochemical analysis of wastewater
all samples were below the permissible limit from Mehkar city:
and ranging from 1.0870.23 mg/L to
2.050.11 mg/L. The physicochemical results of municipal
wastewater were given in table 3. Total five
Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

samples were collected from Mehkar at The BOD and COD values of samples were
alternate days. The pH of the five samples ranging from 13011.08 mg/L to 23835.04
was within the permissible limit, ranging mg/L and 27824.11 mg/L to 48038.11
from 6.70.05 to 7.10.01. The minimum mg/L respectively. The ammonical nitrogen
value of BOD was 729.33 mg/L whereas was ranging from 414.87 mg/L to 668.91
maximum value was 121.511.66 mg/L. mg/L.
The BOD values of all samples were higher
than permissible limit. The COD was The two samples showed high value of
ranging from 1258.32 mg/L to 19415.48 ammonical nitrogen than permissible limit
mg/L. The COD values of all samples were whereas three showed less value than
within the permissible limit. The ammonical permissible limit. The nitrate was ranging
nitrogen was ranging from 28.53.06 mg/L from 1298.82 mg/L to 18818.25 mg/L.
to 674.47 mg/L. Out of the five samples, All the five samples showed higher values
two samples showed higher concentration of of nitrate than permissible limit. The
ammonical nitrogen than permissible limit minimum concentration of Phosphate was
and they were 674.47 mg/L and 66.58.77 1.450.21 mg/L whereas maximum
mg/L. The remaining three samples values concentration was 2.090.11mg/L. These
showed in table 3 were below the phosphate values were within the
permissible limit. permissible limit.

Physicochemical analysis of wastewater The sampling for physico-chemical analysis

from Khamgaon city: was done at morning from 8.00 to 10.0 am
because maximum house holding activities
The results of physicochemical analysis of are occurring at morning than afternoon and
municipal wastewater from Khamgaon were evening. The physicochemical analysis of
given in table 4. The total five samples were four cities reveals that pH of municipal
collected at alternate days. All the five wastewater ranging from 6.7 to 7.2. These
samples showed pH value within the pH values were within the permissible limit
permissible limit, ranging from 6.70.02 to in accordance with World Health
7.10.12. The BOD and COD values of all Organization standards.
samples were above the permissible limit.

Table.1 Physico-chemical characteristics of municipal wastewater from Buldana

Sr. No. pH BOD COD Ammonical Nitrate Phosphate

(mg/L) (mg/L) Nitrogen(mg/L) (mg/L) (mg/L)
1 6.90.08 14418 26422.27 4614.18 15314.11 1.0870.23
2 7.20.02 12022.91 18819.05 6712.0 96.513.44 1.870.35
3 6.80.01 769.96 13413.0 55.519.37 13210.39 1.460.11
4 6.90.02 11010.21 21410.51 484.0 156.50.5 1.340.04
5 7.00.12 826.93 1561.73 66.510.04 1665.0 2.050.11
= Standard Deviation

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

Table.2 Physico-chemical characteristics of municipal wastewater from Chikhli

Sr. No. pH BOD COD Ammonical Nitrate Phosphate

(mg/L) (mg/L) Nitrogen(mg/L) (mg/L) (mg/L)
1 6.80.2 12722.08 26116.34 385.05 8612.2 1.4530.05
2 6.80.11 988.2 15624.52 464.54 12224.66 1.220.02
3 6.90.02 12416.04 21512.54 346.8 14514.0 1.320.11
4 6.80.04 13610.2 22025.14 541.2 944.8 1.010.08
5 6.80.01 785.88 1159.55 433.42 44.55.5 2.10.14
= Standard Deviation

Table.3 Physico-chemical characteristics of municipal wastewater from Mehkar

Sr. No. pH BOD COD Ammonical Nitrate Phosphate

(mg/L) (mg/L) Nitrogen(mg/L) (mg/L) (mg/L)
1 7.10.01 729.33 18728.55 674.47 13416.22 1.540.05
2 7.00.01 10810.12 18020.15 66.58.77 14513.55 1.040.34
3 7.00.04 121.511.66 19415.48 342.08 21026.11 1.820.14
4 6.80.01 888.90 1258.32 42.85.41 120.59.65 2.080.4
5 6.70.05 95.54.85 160.518.14 28.53.06 13010.55 0.980.02
= Standard Deviation

Table.4 Physico-chemical characteristics of municipal wastewater from Khamgaon

Sr. No. pH BOD COD Ammonical Nitrate Phosphate

(mg/L) (mg/L) Nitrogen(mg/L) (mg/L) (mg/L)
1 6.70.02 23835.04 42334.86 414.44 1298.82 2.040.03
2 7.00.0 19016.78 48038.11 566.08 13115.6 1.660.12
3 7.10.12 15527.0 31518.16 668.91 1767.65 2.090.11
4 6.90.14 13011.08 27824.11 481.66 14412.14 1.880.05
5 6.90.02 21015.15 38025.52 414.87 18818.25 1.450.21
= Standard Deviation

The permissible limit for pH is 6.00 to 9.00 organic load present in wastewater by
of wastewater which must be discharged various house holding activities as higher
into the sea or environment (WHO, 2006). population of the city than other cities.
The pH values of our study were similar to
Shirin and Yadav (2014). The BOD and Kandhasamy and Santhaguru (1994)
COD were ranging from 72 mg/L to 238 reported that the higher BOD may be due to
mg/L and 115 mg/L to 480 mg/L higher organic load. In our study, BOD and
respectively. The wastewater samples from COD values were similar to Dubey (2013)
Khamgaon city showed higher BOD and and Shirin and Yadav (2014). The nitrate
COD values as compared to Buldana, was ranging from 44.5 mg/L to 210 mg/L.
Chikhli and Mehkar city. The probable Our nitrate values were similar to Singh et
reason for the higher BOD and COD is high al., (2012) and higher than Dubey (2013).

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 577-584

The phosphate values were ranging from Das, A.C., Baruah, B.K., Baruah, D.,
0.98 mg/L to 2.1 mg/L. In present study, our Sengupta, S. 2003. Study on wetlands
phosphate values were similar with of Guwahati city: Two water quality
Sarkinnoma et al., (2013) and Sonune et al. of rivers and drains. Poll. Res., 22(1):
(2015). The ammonical nitrogen was 117 119.
ranging from 28.5 mg/L to 67 mg/L. Similar Dubey, S., 2013. Analysis of physico-
results were showed by Binu Kumari et al. chemical parameters of Kshipra river
(2006). The high organic load and its water at Ujjain, India. Int. Res. J.
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The present study reveals that municipal producing bacterium from domestic
wastewater from Buldana district contains wastewater of Buldana District
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