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1
INTRODUCTION
Water is an essential natural resource for sustaining life and our environment on this
earth. Water is always available in abundance on this planet. Water is not only vital to life but it
is also a vital component of healthy functioning of any ecosystem (Simmons, 1999) as it is in
continuous interaction with the surrounding air and land and living things. Water is also
geologically important because of its role in weathering, erosion, transportation and deposition of
sediment (The Atlas of Canada, 2004). Unfortunately, unless we use our water wisely, the
water bodies such as rivers, lakes, and groundwater etc. can become depleted or polluted, and
unavailable or unsuitable for life. Water is not only the survival resource of all living beings but
also the main vector for all development activities and is integratedly related with all ecological
and societal processes (Viessman and Hammer, 1998).
The water resources are being utilized for drinking, irrigation and industrial purposes.
There is growing concern on deterioration of water quality due to geogenic and anthropogenic
activities. The quality of water has undergone a change to an extent that the use of such water
could be hazardous. Increase in overall salinity of the ground water and/or presence of high
concentrations of fluoride, nitrate, iron, arsenic, total hardness and few toxic metal ions have
been noticed in large areas in several states of India.
Jaipur district with geographical area of 11,061.44 sq. km forms east-central part of the
Rajasthan State is also popularly known as Pink city and is situated towards central part of the
district. Jaipur is very much on the world tourist map, known for gem & jewelry and is also
popular for Sanganer & Bagru prints. In the present study Sanganer town is selected as study
area. Sanganer town situated 20 km away from the main city of Jaipur is located south of Jaipur
and lying 260 49’-26051’N and 750 46’-750 51’ E.
STATUS OF WATER SUPPLY AND DEMAND
At the time of foundation of Jaipur city in 1727, water supply scheme started by
construction of Jhalaras (a big public well) in each of the nine Chowkaris (squares). In the
beginning of the 18th century the citizens used to draw water from 100 open wells and baoris
located in the city. The first water supply scheme for City Palace where water from open wells
was drawn with oxen and was supplied through a small canal. In the mid of 18th
century the
2
walled city was supplied water from Amanishah Nala through canals up to Chhoti Chopar and
Bari Chopar from where people used to take water. Piped-water supply to Jaipur for public began
in 1874 with the construction of a Reservoir of 4.5 MLD capacity across Amanisha Nala. Open
wells were constructed in the Nala to meet the drinking water demand. Later on the Ramgarh
dam was constructed in the year 1906. Due to increased population a scheme was taken up to
augment water supply from the lake and closed pipe lines were constructed for supply to the
whole city. Amanishah nala was then declared as a waste water channel. But due to various
usages of its water i.e. printing, dying, irrigation, which in a huge amount happens in
SANGANER area has become a serious problem and it is a source of water pollution.
Sanganer is very famous for a special type of printing known as “Sanganeri Printing”
the process involves the use of various kinds of chemical dyes such as rapid indigo, direct
aniline black, which also includes many metal based dyes used for fastening colors. There are
around 700 varieties of dyes and dye intermediates produced in India, mainly direct dyes, acid
dyes, reactive dyes and pigments. Most of these dyes have not been characterized regarding their
chemical nature, purity, possible toxicity or their impact on health and the environment. Yet,
they are widely used by textile, leather, paint and even the food industry.
Fig.1 Sanganer, south east of Jaipur Rajasthan
3
Printing involves use of large amount of water and thus large quantity of waste water is
also generated. The untreated sewage water and waste water from textile industries (which
contains variety of chemicals such as Aniline, Caustic soda, Acids, Bleaching powder etc.
including heavy metals) is used in irrigating agricultural fields located in Amanishah nala, for
growing vegetables and other crop plants. It certainly makes the part of food chain. The wastes
released from such industries cause soil, surface and ground water pollution, besides causing a
number of adverse effects on agricultural products, animals and health of people living in that
area as human beings in and around Sanganer in Jaipur district consume these vegetables and
products of other crop plants.
Fig. 2 Textile and Dying industries Fig.4 Water used in Irrigation
Rapid industrialization of textile and dyeing industry in the world pose a major
environmental threat because of the large amounts of water and dyes involved in the
manufacturing process. Large amounts of chemically different dyes are employed for various
industrial applications including textile dyeing. Dye production in India is estimated to be around
64,000 tones, which is about 6.6% of the world production. The textile industry in India alone
consumes up to 80% of the total dyestuffs produced. In Rajasthan state particularly, textile mills
represent an important economic sector. Sanganer is famous for dyeing and printing of colorful
dresses, bed sheets, curtains, dress material and variety of other textiles.
Most of the industries have been using many dyes as mentioned but the agony despite
the best efforts made, is that there is no common effluent treatment plant installed in Sanganer.
The untreated industrial wastewater along with untreated domestic sewage may be seen
accumulated in many areas in absence of proper drainage Bulk of the textile products of these
industries is exported. It is located about 15 km south of Jaipur, the State capital that has a
4
population of more than two million people. The total area of Sanganer is about 635.5 Sq. km out
of which, 12.9 Sq. km comprises the urban area. Most of the textile industries of Sanganer are
concentrated in this urban area. There are estimated to be around 500 block and screen printing
units in Sanganer. Among the total dyestuff consumption, it has been reported that textile
industry accounts for 67% of the total dyestuff market. These activities often lead to alteration of
water quality by raising the physio-chemical parameters above the allowable limits and
ultimately results into Environmental pollution.
Environmental pollution is one of the most horrible crises that we are facing today. Due
to the increased urbanization and industrialization surface water pollution has become an crucial
problem. It is necessary to obtain precise and appropriate information to observe the quality of
any water resources and the development of some useful tools to keep watch on the quality of
such priceless water resources to retain their excellence for various beneficial uses.
Water Pollution in Amanishah nala:
Increasing industrialization and urbanization have caused not only air, sound and surface water
pollution but have also caused ground water pollution in the Jaipur city resulting in adverse
effects on the health. Voluminous liquid wastes are generated by the dyeing and printing industry
and are disposed off in carrier channels (canals). In addition, some industrial units are also
pouring their effluents into the Amanishah Nala. These liquid wastes are also being used for
irrigation purposes. The unused part of effluent water is allowed to accumulate near the bunds in
the peripheral areas giving adequate time period to this effluent water to percolate and reach the
saturated zone. Thereby degrading and deteriorating ground water quality. The polluted surface
water flows as per hydraulic gradient and gets concentrated at favorable geomorphic locations
where its flow is sluggish. There is an urgent warning and calls for measures to tackle the quality
hazards of ground water. It is therefore recommended that the liquid effluents should be treated
and beneficiated to remove the hazardous constituents before their disposal and also to
encourage / motivate to use vegetable dyes. Alternatively, the dyes having higher concentration
of fluoride should be replaced by alternative dyes. It is also recommended to develop a proper
sewerage network system in urban agglomerate areas particularly in the walled city so as to
prevent mixing with ground water.
5
Parameters Reason for the analysis
Physical Parameters
Temperature Temperature can exert great control over aquatic communities.
If the overall water body temperature of a system is altered, an
aquatic community shift can be expected.
In water above 30o
C, a suppression of all benthic organisms
can be expected. Also, different plankton groups will flourish
under different temperatures. For example, diatoms dominate
at 20 - 25 degrees C, green algae dominate at 30 - 35 degrees
C, and cyano-bacteria dominate above 35 degrees C.
Conductivity Conductivity indicates the presence of ions within the water,
usually due to in majority, saline water and in part, leaching. It
can also indicate industrial discharges.
The removal of vegetation and conversion into monoculture
may cause run-off to flow out immediate thus decrease
recharge during drier period. Hence, saline intrusion may go
upstream and this can be indicated by higher conductivity.
Chemical Parameters
pH value pH is an indicator of the existence of biological life as most of
them thrive in a quite narrow and critical pH range.
Salinity High salinity may interfere with the growth of aquatic
vegetation. Salt may decrease the osmotic pressure, causing
water to flow out of the plant to achieve equilibrium. Less
water can be absorbed by the plant, causing stunted growth
and reduced yields. High salt concentrations may cause leaf tip
and marginal leaf burn, bleaching, or defoliation.
As per Conductivity, salinity (NaCl content, g/kg) can be used
to check for possible saline intrusion in future.
Total Dissolved Solids, TDS The total dissolved solids (TDS) in water consist of inorganic
6
salts and dissolved materials. In natural waters, salts are
chemical compounds comprised of anions such as carbonates,
chlorides, sulphates, and nitrates (primarily in ground water),
and cations such as potassium (K), magnesium (Mg), calcium
(Ca), and sodium (Na). In ambient conditions, these
compounds are present in proportions that create a balanced
solution. If there are additional inputs of dissolved solids to the
system, the balance is altered and detrimental effects may be
seen. Inputs include both natural and anthropogenic source.
Organic constituents
Dissolved Oxygen (DO) DO is essential for aquatic life. A low DO (less than 2mg/l)
would indicate poor water quality and thus would have
difficulty in sustaining many sensitive aquatic life.
Biochemical Oxygen Demand,
BOD
BOD is a measure of organic pollution to both waste and
surface water. High BOD is an indication of poor water
quality. For this tree plantation project, any discharge of waste
into the waterways would affect the water quality and thus
users downstream.
Chemical Oxygen Demand,
COD
COD is an indicator of organics in the water, usually used in
conjunction with BOD.
High organic inputs trigger deoxygenation. If excess organics
are introduced to the system, there is potential for complete
depletion of dissolved oxygen. Without oxygen, the entire
aquatic community is threatened. The only organisms present
will be air- breathing insects and anaerobic bacteria.
If all oxygen is depleted, aerobic decomposition ceases and
further organic breakdown is accomplished anaerobically.
Anaerobic microbes obtain energy from oxygen bound to other
molecules such as sulphate compounds. Thus, anoxic
conditions result in the mobilization of many otherwise
7
insoluble compounds.
In areas of high organics there is frequently evidence of rapid
sewage fungus colonization. Sewage fungus appears as slimy
or fluffy cotton wool-like growths of micro-organisms which
may include filamentous bacteria, fungi, and protozoa such as
Sphaerotilus natans, Leptomitus lacteus, and Carchesium
polypinuym, respectively. The various effects of the sewage
fungus masses include silt and detritus entrapment, the
smothering of aquatic macrophytes, and a decrease in water
flow velocities. An accumulation of sediment allows a shift in
the aquatic system structure as colonization by silt-loving
organisms occur. In addition, masses of sewage fungus may
break off and float away, causing localized areas of dissolved
oxygen demand elsewhere in the water body.
Organic levels decrease with distance away from the source. In
a standing water body such as a lake, currents are generally not
powerful enough to transport large amounts of organics. In a
moving water body, the saprotrophic organisms (organisms
feeding on decaying organic matter) break down the organics
during transportation away from the source. Hence, there is a
decline in the oxygen demand and an increase of dissolved
oxygen in the water. Community structure will gradually
return to ambient with distance downstream from the source.
Microbiological
Total Coliform Count Microbiological test is to detect the Level of pollutions caused
by living thing especially human who live or work in the area
especially upstream of the site.
These tests are based on coliform bacteria as the indicator
organism. The presence of these indicative organisms is
evidence that the water has been polluted with faeces of
humans or other warm-blooded animals.
Faecal Coliform Count
8
Parasitological Parasitology is concerned with the study of parasites, their
hosts and their relationship with one another.
By performing parasitological analysis we search for formed
cellular elements, casts, bacteria, yeast, parasites and crystals
in centrifuged water sample
Table 1 Water Quality Parameters and Definitions
Objective of dissertation:
Amanishah nala water is highly contaminated and microorganisms rich, which is very
much harmful to human health. My objectives during this project are as follows:
Conduction of Physical analysis,
Chemical analysis,
Microbiological analysis
Parasitological Analysis
These analyses will be done on Amanishah Nala water, Sanganer, Jaipur. So that
suitability of the water for farming and any contamination can be measured.
9
REVIEW OF LITERATURE
Systematic Hydro geological Survey in the entire district was completed by
Ground Water Wing of Geological Survey of India & Central Ground Water Board
which further has been reappraised periodically during the Annual Action Plans 1986-87
(7 blocks), 1992-93 (3 blocks), 2004-05 & 2005-06. Two Mass Awareness Programmes
and five Water Management Training Programmes have been conducted. World Water
Day is also often celebrated in Jaipur.
Literature on studies on the impact of waste water on agricultural crops reveal that
crop plants and vegetables grown in the agricultural fields by using untreated waste water
were adversely affected both qualitatively and quantitatively.
Ganeshan and Manoharan (1983) studied the effect of cadmium and mercury on
germination of seeds, growth of seedling and matter production of Abelmoschus esculents
and found that cadmium is more toxic than mercury and the water of high concentration
of these pollutants can only be utilized for irrigation by proper dilution.
Azad et al. (1984) studied the impact of the lake water on crop plants and surrounding
populations and measured the levels of various physical and chemical parameters.
Brown and Wilkins (1986), Dayama (1987) studied influence of dyeing and textile
waste water on nodulation and germination of Cicer aeritium.
Rana and Masood (2002) conducted a pot study to investigate the toxic effects of certain
heavy metals on the plant growth and grain yield of wheat (Triticum aestivum L.).
S. K. Sharma, (2004) evaluated groundwater samples from Jaipur, Rajasthan, India to
determine their suitability for irrigation and drinking purposes. He found that the quality
of almost all samples was within permissible limits but contents of EC, sodium, nitrate,
TDS and DO were not within permissible limits. On the other hand, the general
10
characteristic of the samples could be classified under moderate category and were good
for household, irrigation and commercial purposes.
BK Nayak, BC Acharya, UC Panda et al (2004) studied water quality parameters for
the entire Chilka lake covering a maximum of 23 sampling stations and found that pH of
water was alkaline throughout the lake and both pH and salinity varied widely. Higher
pH with low salinity zones reflected disintegration of submerged weeds. Correlation
analysis supported the increase of pH, high photosynthetic activity, high nutrients as well
as phosphate depletion due to phytoplankton utilization in the fresh water zone.
AP Sawane, PG Puranik et al (2004) studied assessment of pollution status of river Irai
(Dist. Chandrapur) and found increased values of BOD in river water which was
indicative of increased quantity of industrial effluents. The reduced DO content was due
to hot ash slurry from thermal power plant. They also collected data from present study
which reveals that there is is an inverse relationship between DO and BOD and
portability of Irai river water is below the standard permissible limit.
Moti R Sharma, AB Gupta, JK Bassin (2004) stated that the dissolved oxygen in the
stream is below 4mg/l in a stretch of 2600m and therefore water is not fit for public
supply, bathing, fish culture and wildlife in Hathli stream Shivalik Himalayan.
Vijendra Singh, Chandel Singh (2005) analyzed groundwater and wastewater samples
from ‘Amanishah Nala’ and hand pump of seven industrial areas and adjacent localities
of Jaipur city with the help of standard methods of APHA and Black. The values obtained
were compared with standards of ISR, ICMR and WHO. The concentrations of various
parameters are within permissible limits in both groundwater and wastewater but definite
contaminations with special reference to EC, TDS and COD in wastewater have been
observed which calls for at least primary treatment of wastewater before being used for
irrigation.
11
Dinesh Kumar et al (2005) monitored Sanganer nala and surrounding tube wells and
stated that discharge of untreated industrial effluents and sewage in to nala have
contributed considerable pollution in the ground water in its vicinal areas, and is harmful
for use in agriculture and drinking purposes
JD Sharma, P Jain, D Sohu (2005) revealed that pH, EC and alkalinity of all the
samples from villages of Sanganer, Jaipur were very high which can be correlated with
high TDS and chloride. Twenty eight percent villages contained high fluoride
concentration than permissible limit i.e., 1.5ppm. A positive correlation was observed
between pH and fluoride, TDS and EC. Hardness showed negative correlation with
fluoride and pH.
SS Asadi SS, Padmaja Vuppala, M Anji Reddy (2005) stated that high concentrations
of total dissolved solids, nitrates, fluorides and total hardness were observed in few
industrial and densely populated areas in Hyderabad indicating deteriorated water quality
while the other areas exhibited moderate to good water quality.
Bhaskar Bhadra et al (2005) found higher range of alkalinity, ammonia content and
chloride content in Torsa than Kaljani. River Kaljani showed higher COD range than
Torsa. Mean BOD value of both these rivers ranged between 0.93-1.65 mg/l. Overall
TDS content of Kaljani was found to be lower than Torsa. Maximum phosphate content
was observed at the downstream of both the rivers.
Ram Chandra, RN Prasad (2005) studied various water quality parameters related to
the deterioration in water quality of Surya Kund, Lohargal (Rajasthan) during the mass
bathing of religious importance and said that it is necessary to take adequate
precautionary measures to prevent outbreak of any epidemics.
R Rajaram, M Srinivasan, M Rajasegar (2005) said that nutrient concentrations were
higher during monsoon season and low during summer season at two stations of Uppanar
estuary, Cuddalore in relation to effluent discharges from SIPCOT industries. There are
12
44 industries discharging their effluents into Uppanar estuary, which may influence the
biota.
SR Vishnoi, PN Srivastava (2005) collected water sample from three different sites of
river Jojari at Salawas, Jodhpur and carried hydro biological studies. They found that
the pH, chloride, salinity, total alkalinity, total hardness, dissolved oxygen and TDS were
absolutely higher than the standard values of portable water on account of contamination
of river due to industrial effluents. The river has become unsuitable for the growth and
survivability of aquatic flora and fauna. The pollution impact was found to be
predominant during summer and minimal during monsoon season.
Bhatnagar et al. (2006) reported that waste water effluents from textile dyeing and
printing industries of Sanganer were discharged directly, without any treatment, into
Amanishah nala drainage. The drainage water takes the dissolved toxicants to flora and
fauna, including crops and seasonal vegetables, being grown in the land adjoining the
nala drainage. The mutagenic potential of vegetables irrigated by water of Amanishah
nala drainage was investigated by them.
Marques et al. (2007) studied the levels of zinc accumulated in field conditions, by
roots, stems, and leaves of Rubusulmifolius and Phragmites australis, the species
indigenous to the banks of a stream in a Portuguese contaminated site.
Oporto et al. (2007) studied elevated Cd concentration in potato tubers due to irrigation
with river water contamination by mining in Potsi, Bolivia.
Nupur Mathur, Pradeep Bhatnagarit(2007) collected water samples from the
agricultural fields of Trigonella foenumgraecum grown in Sanganer area. These samples
were found to contain pH in the range from 8.4 to 11.9. Electric conductivity ranged from
0.564 to 3.203mmhos/cm, whereas concentrations of total dissolved solids were found in
the range from 750 to 4670 mg/L and Chlorides from 315.40 to 1304.60 mg/L
respectively. The data indicate that effluent concentrations in the waste water of
Amanishah nala have considerably been increased during the past few decades.
13
Sikka et al. (2008) stated that the disposal of industrial and sewage water was a problem
of increasing importance throughout the world. In India and most of the developing
countries untreated sewage and industrial wastes are discharged on land or into the
running water streams which is used for irrigating crops.
The objectives of these scientific investigations has been to determine the agro-chemistry
of the water and to classify the waste water in order to evaluate the effect of pollutants on
plants, and the water suitability for irrigation uses and the present report is also aimed at
it. The analysis of waste water is made at pre-flowering, flowering and post flowering
stages in order to make comparison of the quality of water at these stage sand in turn
correlate with external, internal morphological changes, if any, at the three stages of
growth.
14
MATERIAL AND METHODS
Two samples INDUSTRIAL EFFLUENT and WASTE WATER were collected from
Amanishah nala and Physical, Chemical, Bacteriological and Parasitological analysis
were performed following the standard protocols.
Fig.4 Waste water sample Fig. 5 Industrial effluent water sample
1. Physical analysis of water
EXPERIMENT 1
Objective: To measure the temperature of the given water sample.
Requirements: Thermometer, glass ware, water sample.
Procedure:
1) Take 50 ml Water sample in beaker
2) Wipe and clean the thermometer with blotting paper and immerse in
sample.
3) Still the sample well before noting down the temperature.
4) Note the constant reading.
15
EXPERIMENT 2
Objective: To measure the conductivity of the given water sample.
Requirements: Conductivity meter, water sample.
Procedure:
1) Calibrate the conductivity meter.
2) Check the conductivity of water sample.
2. Chemical analysis of water
EXPERIMENT 1
Objective: To measure the pH of the given water sample.
Requirements: pH meter, buffer solution of 4 and 7, water sample.
Procedure:
1) Calibrate the pH meter.
2) Check the pH of the water sample.
EXPERIMENT 2
Objective: To estimate the salinity (Cl-) in the given water sample.
Requirements: Glassware, burette, pipette, conical flask, dropper.
Reagents: AgNO3 (0.02 N), Potassium chromate as indicator (5%)
Dissolve 0.34 gm AgNO3 in 100 ml DW
Dissolve 2.5 gm Potassium chromate in 50 ml DW.
16
Theory: NaCl is the main substance responsible for chloride conc. in water. Chloride is
estimated on the basis of argentometric method in high chloride is precipitated with
potassium chromate.
Permissible limit for chloride is 250 mg.
AgNO3 + Cl- AgCl (white ppt.) + NO3
-
2 AgNO3 + K2CrO4 Ag2CrO4(Brick Red) + 2KNO3
Procedure:
1) 50 ml of water sample is taken and 2 ml of potassium chromate is added
as an indicator.
2) Stir well and titrate with AgNO3.
3) End point is the color change from yellow to brick red, note it down.
Observation:
Sample 1 2 3
Industrial effluent 16.7 16.3 16.3
Waste water 20 20.3 20
Calculation: Concentration of Cl-(mg/l) = (V x N x 1000)/ volume of sample
Where,
V = Volume of titrant
N =Normality of titrant.
Industrial effluent: (16.3 X 0.02 X 1000)/ 50 = 6.52
Waste water: (20 X 0.02 X 1000)/ 50 = 8
17
EXPERIMENT 3
Objective: To determine the acidity of the given water sample.
Reagents: NaOH (0.05 N) dissolve 1gm in 500 ml DW, Methyl orange indicator,
Phenolphthalein indicator
Theory: Acidity of water can be neutralized by strong base. In natural unpolluted fresh
water, acidity is mostly due to presence of free CO2 in the form of carbonic acid.
Acidity can be determined by titrating sample strong base like NaOH, using
methyl orange and phenolphthalein as indicators. If sample has strong mineral acid and
its salt just titrates to pH 3.7, using methyl orange as an indicator then it is called methyl
orange acidity.
If sample is titrated to pH 8.3 then phenolphthalein indicator should be used. The
resulting value is called total acidity.
Result of acidity should always be mentioned with pH titration method is suitable mainly
for samples which are colorless.
Procedure:
1) 50 ml water sample is taken and 2-3 ml of methyl orange was added.
2) If the solution turns yellow, that shows methyl orange acidity is absent, i.e.
pH is more 3.7.
3) If solution turns red, it is titrated with 0.05 N NaOH till it becomes yellow.
Note the end point.
4) 2-3 drops phenolphthalein indicator is added to the sample.
5) Sample is further titrated with NaOH until the content turns pink.
Calculations:
When, A= Vol. of NaOH used with methyl orange in titrating the sample,
B= Vol. of NaOH used with phenolphthalein in titrating the sample to pH 3.7 – 8.3.
18
i. Methyl orange acidity: (A X N X 1000 X 50)/ vol. of sample
Industrial effluent= 0
Waste water= 0
ii. Phenolphthalein acidity: (B X N X 1000 X 50)/ vol. of sample
Industrial effluent= (4.1 X 0.05 X 1000 X 50)/ 50 = 205
Waste water= (2.5 X 0.05 X 1000 X 50)/ 50 = 125
iii. Total acidity: Methyl orange acidity + Phenolphthalein acidity
Industrial effluent: 0 + 205 = 205
Waste water: 0 + 125 = 125
EXPERIMENT 4
Objective: To estimate the total alkalinity (carbonate and bicarbonate) in the given water
sample.
Requirements: Glassware: conical flask, beaker, dropper,
Reagents: HCL(0.1N), Methyl orange, Phenolphthalein indicator.
Theory: Total alkalinity is the measurement of CO3-, HCO3
- and OH
-. The alkalinity in
water is generally imparted by salt of carbonates, bicarbonates, phosphates, nitrate,
silicate etc. together with hydroxyl ion in Free State. Most of the water is rich in
carbonate, bicarbonate with little concentration of other alkalinity imparting ions.
Total alkalinity, carbonates, bicarbonates can be estimated by filtering the sample
with strong acid either HCL or H2SO4.
First it is titrated to pH 8.3 using phenolphthalein as an indicator then further pH
between 4.2- 5.4 with methyl orange indicator. In first, the value is called
phenolphthalein alkalinity and in second is total alkalinity. Value of carbonate,
bicarbonate and hydroxyl ion can completed from these two types of alkalinities.
19
Procedure:
1) 50 ml of sample is taken in a flask and 2 drops of phenolphthalein
indicator.
2) If the solution remains colorless, that shows phenolphthalein alkalinity is
absent.
3) If solution turns pink after adding phenolphthalein then titrate it with 0.1
N HCl until the color disappears at the end point. Note the end point.
4) Now, 2-3 drops of methyl range are added to the same sample and
continue the titration until the yellow color changes to pink. Note down
the point.
Calculations:
When A= HCl used with only phenolphthalein,
B= HCl used with only methyl orange.
i. Methyl orange acidity: (A X N X 1000 X 50)/ vol. of sample
Industrial effluent= (4.1 X .1 X 1000 X 50) 50 = 420
Waste water= (6.3 X .1 X 1000 X 50) 50 = 630
ii. Phenolphthalein acidity: (A X N X 1000 X 50)/ vol. of sample
Industrial effluent = 0
Waste water = 0
iii. Total acidity: Methyl orange acidity + Phenolphthalein acidity
Industrial effluent = 420 + 0 = 420
Waste water = 630 + 0 = 630
20
EXPERIMENT 5
Objective: Determination of total dissolved solids of water.
Requirements: Evaporating dish, hot water bath, desiccators, Whatmann filter papers.
Theory: Water, the universal solvent has large no. of salts dissolved in it which largely
govern the physico- chemical properties of water and in turn have an indirect effect on
the flora and fauna. Total dissolved solids (TDS) are determined as the residue left after
the evaporation of the filtered sample.
Procedure:
1) Take the weight of the evaporating dish.
2) Filter the sample of suitable quantity through Whatmann filter paper.
3) Transfer the sample to the evaporating dish.
4) Evaporate water completely.
5) Note the weight of the dish along with the content after cooling in a
desiccators.
Calculations: TDS = {(B-A) X 106} / V
Where, A=Initial weight of the dish,
B=Final weight of the dish,
V= volume of the water sample taken.
Industrial effluent = {(100.05 - 100) X 106}/ 100 = 500
Waste water {(102.13 – 102.04) X 106}/ 100 = 700
21
3. Organic constituents in water
EXPERIMENT 1
Objective: To estimate free CO2 in the given water sample.
Requirements: Glassware: Burette, pipette, funnel, flask
Reagents: Sodium Hydroxide (NaOH 0.05 N), Phenolphthalein indicator.
Procedure:
1) 50 ml of sample is taken(without bubbling).
2) 2-3 drops of phenolphthalein is added. Change of color to pink indicates
absence of free CO2.
3) If the sample remains colorless, titrate it with 0.05 N NaOH till pink color
appears.
Calculation: Free CO2 (mg/ml) = (S x N x 1000 x 44)/volume of sample
Industrial effluent: (1.7 X 0.05 X 1000 x 44)/ 50 = 74.8
Waste water: (1.1 X 0.05 X 1000 x 44)/ 50 = 48.4
22
EXPERIMENT 2
Objective: Determination of dissolved oxygen (DO) of water.
Requirements: Water sample (tap water), Sodium thiosulphate (0.025 N), (dissolve 1.24
gm in 50 ml distilled water), Magnous sulphate (40 gm in 100 ml distilled water),
Alkaline potassium iodide solution (10 gm KOH and 5 gm KI in 20 ml boiled distilled
water), Starch indicator (1%), concentrated sulphuric acid, DO bottles.
Theory:
Dissolved oxygen of water is of paramount importance to all living organisms. The
presence of DO in water may be mainly attributed to two distinct phenomenons:
1. Direct diffusion from the air.
2. Photosynthetic evolution by aquatic autotrophs.
The first one is purely a physical process and depends on the solubility of oxygen under
the influence of temperature, salinity, water movements etc. whereas the later is a
biological process and depends on availability of light and the rate of metabolic processes
resulting in diurnal fluctuations.
The estimation of DO is done by titrimetric method. The oxygen of the water combines
with manganous hydroxide, which on acidification liberates iodine equivalent to that of
oxygen fixed. This iodine is titrated by standard sodium thiosulphate solution using
starch as indicator.
Procedure:
1) Collect the water sample without bubbling in the 250 ml glass bottle. Add
2 ml each of magnous sulphate and alkaline potassium iodide solutions in
succession, right at the bottom of bottle with separate pipettes and replace
the stopper.
2) Shake the bottle in upside down direction at least 6 times.
3) Allow the brown precipitate to settle.
23
4) Add 2 ml of concentrated sulfuric acid and shake the Stoppard bottle to
dissolve the brown precipitates.
5) Take 25 ml of sample in a flask and titrate with thiosulfate solution (taken
in the burette) till the color changes to pale straw.
6) Add 2 drops of starch solution to the above flask which changes the color
of the contents from pale to blue.
7) Titrate again with thiosulfate solution till the blue color disappears.
Calculations:
DO (mg/l) = 8 x 1000 x N x v / V
Where,
V = Volume of sample taken
v = Volume of titrant used
N = Normality of the titrant.
24
EXPERIMENT 3
Objective: Determination of biochemical oxygen demand (BOD) of water.
Requirements: Water sample (tap water), Reagents for DO estimation, BOD bottles,
flask, pipette, BOD incubator, pH meter.
Theory:
The BOD is a way of expressing the amount of organic compounds in sewage as
measured by the volume of oxygen required by bacteria to metabolize it under aerobic
conditions. It is good index of the organic pollution. If the amount of organic matter in
sewage is more, the more oxygen will be utilized by bacteria to degrade it. Dumping
sewage that contains high BOD increases the concentration of soluble organic
compounds in the aquatic body where it is discharged. Digestion of these organic
compounds in natural ecosystems, such as lakes, rivers, can deplete available O2 and
resulting in asphyxiation of fish.
The BOD of a water sample is generally measured by incubating the sample at 20
degrees for five days in the dark under aerobic conditions. In tropical and sub-tropical
belts, where the temperature and rate of metabolic activities are higher, the incubation
should preferably be done at 27 degrees for 3 days.
Procedure:
1) Fill the water sample in duplicate numbers in BOD bottles without
bubbling.
2) Determine dissolved oxygen content in one set of each sample by the
titration method.
3) Incubate the rest of the bottles at 27 degrees in a BOD incubator for 3
days.
4) After 3 days estimate the oxygen concentration in all the three incubated
samples.
5) Take the readings.(D2)
25
Calculations:
BOD (mg/l) = D1 – D2
Where,
D1 = Initial DO of sample.
D2 = DO after 3 days incubation.
26
EXPERIMENT 4
Objective: Determination of chemical oxygen demand (COD) of water sample.
Requirement: Water sample, potassium dichromate solution (0.1 N), sodium
thiosulphate (0.1 M), Sulphuric acid (2M), potassium iodide solution (10%), water bath,
titration assembly, 100 ml conical flask, water blanks.
Theory: In recent times, with the increase of pollution by discharging large amounts of
various chemically oxidizable organic substances of different nature entering the aquatic
systems, BOD alone doesn’t give a clear picture of the organic matter content of the
water sample.
In addition, the various toxicants in the sample may severely affect the validity of the
BOD test. Hence chemical oxygen demand is a better estimate of the organic matter,
which needs no sophistication and is time saving. However, COD, that is oxygen
consumed doesn’t differentiate the stable organic matter from the unstable form.
Therefore, the COD values are not directly comparable to that of BOD.
The amount of organic matter in water is estimated by their oxidability by a chemical
oxidant such as potassium permanganate or potassium dichromate. In the permanganate
method, the organic matter is first oxidized with a known volume of KMnO4 and then
excess of oxygen is allowed to react with potassium iodide to liberate iodine in amounts
equal to the excess oxygen, which is estimated titrimetrically with sodium thiosulphate
solution using starch as an indicator.
Procedure:
1) Take three 100 ml conical flasks and pour 50 ml of water sample in each.
Simultaneously run distilled water blank standards.
2) Add 5 ml of potassium dichromate solution in each of the six flasks.
3) Keep the flasks in water bath at 100 degrees (boiling temperature) for one
hour.
4) Allow the samples to cool for 10 minutes.
5) Add 5 ml of potassium iodide in each flask.
27
6) Add 10 ml of H2SO4 in each flask.
7) Titrate the contents of each flask with 0.1 M sodium thiosulphate until the
appearance of pale yellow color.
8) Add 1 ml of starch solution to each flask (solution turns blue).
9) Titrate it again with 0.1 M sodium thiosulphate until the blue color
disappears completely.
Observations:
Sample 1 2 3
Industrial effluent 4.4 4.4 4.2
Waste water 4.2 4.2 4.3
Blank 4.1 4 4
Calculation: COD of sample (mg/l) = {8 x C x (B-A)} / S
Where,
C = Concentration of titrant
A = Volume of titrant used for blank
B = Volume of titrant used for sample
S = Volume of water sample taken
Industrial effluent: {8 x .1 x (4.4-4)} / 20= 0.016
Waste water: {8 x C x (4.2-4)} / 20 = 0.008
28
4. Bacteriological analysis of water
EXPERIMENT 1
Objective: Bacteriological examination of potable water by most probable number
(MPN) tests.
Theory: Multiple tube fermentation test or most probable number (MPN) test is the most
often used technique for the sanitary analysis of water. The test is used to detect
coliforms that make up approximately 10% of the intestinal microbes of humans and
other animals and have found widespread use as indicator organism of fecal
contamination.
The test is performed sequentially in three stages: presumptive, confirmed and
completed test. Lactose broth tubes are inoculated with different water volumes in the
presumptive test. Tubes that are positive for gas production are inoculated into brilliant
green lactose bile broth in the confirmed test and positive tubes are used to calculate the
MPN of coliforms in the water sample following the statistical table.
The completed test, involving the inoculations of EMB agar plate, nutrient agar
slant and brilliant green lactose bile broth and preparation of a gram stain slide from NA
slant, is used to establish that coliforms bacteria are present in the sample. The complete
process, including the confirmed and completed tests requires at least 4 days of
incubations and transfers.
29
PRESUMPTIVE COLIFORMS TEST:
The presumptive Coliforms test is used to detect coliforms in a water sample. In this test
lactose fermentation tubes are inoculated with different water volumes and production of
acid and gas from the fermentation of lactose in any of the tubes is a presumptive
evidence of coliforms in the water sample.
The lactose broth used in this test is selective for the isolation of coliforms
because of the addition of the bile and lauryl sulphate or brilliant green. A pH indicator
such as Bromocresol purple is also added to lactose broth for the detection of acid. The
color of the indicator changes to yellow with the production of acid from lactose.
Requirements: Water sample (100 ml), Lactose broth medium, Durham tubes (15), 10
ml double strength lactose broth tubes (LB2X) (5), 5 ml single strength lactose broth
tubes (LB1X) (10), Sterile pipettes, one each of 10 ml, 1 ml and 0.1 ml capacity, Bunsen
burner, Mechanical pipetting device, Glass marker pencil.
Procedure:
1) Collect water sample.
2) Pour double strength media 10 ml in 5 tubes, single strength media 9.9 ml
in 5 tubes and 9 ml in 5 tubes.
3) Label 5 double strength lactose broth tubes “10” and 5 single strength
broth tubes “1” another 5 tubes “0.1”.
4) Put media filled Durham’s tubes in inverted position in fermentation tube.
5) Autoclave these tubes.
6) In LF mix the water sample by thoroughly shaking.
7) Aseptically inoculate each “5” tubes (LB2X) with 10 ml of water sample
using 10 ml sterile pipette.
8) Using a 0.1 ml pipette, aseptically inoculate the five tubes (LB1X) with 1
ml of water sample.
9) Using a 0.1 ml pipette, aseptically inoculate the five tubes (LB1X) with
0.1 ml of water sample.
30
10) Incubate all the 15 inoculated tubes aerobically at 35 degree centigrade for
48 hours.
Observations:
Fig.6 Lactose broth tubes with positive test
All 15 tubes +ve in both industrial effluent and waste water
31
CONFIRMED COLIFORMS TEST
This test is used to confirm the presence of coliforms and determine the MPN value in
water sample showing positive or doubtful presumptive test. In the confirmed test, water
samples from all the positive presumptive lactose broth tubes are inoculated into tubes of
brilliant green lactose bile broth and incubated at 35 degree centigrade for 48 hours.
Positive confirmed tubes are used to determine MPN. A statistical method is used to
estimate the population of coliforms, which means that the result obtained in expressed as
the most probable number (MPN) of coliforms. A count of number of lactose
fermentation tubes/brilliant green lactose bile broth showing production of gas following
the incubation period is taken and MPN is found by matching the results with those
provided in the statistical table.
Requirements: 10 ml brilliant green lactose bile broth fermentation tubes (the number
depending upon the tubes showing positive presumptive test), Durham tubes, Inoculation
loop, Bunsen burner.
Procedure:
1) Prepare fermentation tube with 10 ml BGLB media with Durham’s tubes
like previous one.
2) Inoculate brilliant green lactose bile broth tubes with the inoculums from
all lactose broth positive presumptive tubes.
3) Incubate all the inoculated tubes at 35 degree C for 48 hours.
Observations:
Fig.7 Industrial effluent- 15 tube +ve Fig.8 Waste water- 8tubes +ve
32
Table 2 Faecal coliform MPN per 100 ml of sample for three sets of five tubes
containing 1 ml, 0.1 ml and 0.01 ml of sample respectively
33
COMPLETE COLIFORMS TEST
Completed test is used to establish the presence of Coliform bacteria and as confirmatory
test for the presence of E.coli in a water sample. In the completed test, the samples from
the positive brilliant green lactose bile broth from the confirmed test are streaked onto a
selective differential medium for coliforms and inoculated into lactose broth tube as well
as streaked on a nutrient agar plate to perform Gram staining. The medium commonly
used is eosin-methylene blue (EMB) that is selective in nature because of the dye
methylene blue which inhibits the growth of Gram-positive bacteria, allowing the growth
of Gram-negative bacteria EMB is differential in nature in that lactose fermenting
bacteria gives colored colonies (a positive confirmed test) due to the formation of a
complex in EMB that precipitates out onto the coliforms colonies. Non-lactose
fermenters produce colorless colonies on EMB agar. If there is production of acid and gas
in the inoculated lactose broth and there are rod shaped bacteria showing Gram-negative
reaction, these confirm the presence of E.coli in the water sample and are considered a
positive completed test.
Requirements: EMB agar plates, 24 hours coliforms confirm positive brilliant green
lactose bile broth culture (from the confirmed test), 5 ml brilliant green lactose broth
fermentation tube, Nutrient agar slant, Inoculating loop, Bunsen burner.
Procedure:
1) Streak the two EMB agar plates from positive tubes with a sterile
inoculating needle.
2) Incubate the inoculated plates for 24 hrs. at 35 degree C in an inverted
position.
Observations: Green shiny cultures of bacteria with pink purple colony were observed in
positive samples.
Results: Coliforms bacteria are present in water sample. Hence the water sample is not
potable.
Fig.9 Bacterial culture with Industrial effluent:
34
Fig.10 Bacterial culture with Waste water:
Fig.9 10 ml
sample
1 ml sample
0.1 ml sample
10 ml sample 1 ml sample
35
5. Parasitological analysis of water
EXPERIMENT 1
Objective: To identify parasites in water sample under light microscope.
Requirement: water sample, centrifuge, centrifuge tube, slide, cover slip, light
microscope, Lugol’s solution.
Method:
1) 3/4th
of the centrifuge tube was filled with water sample.
2) It was then centrifuged for 5 minutes at 2500 rpm.
3) Supernatant was discarded and 2 drops of Lugol’s solution was added in
the tube with pellet.
4) Tube was shaken until pellet got dissolve.
5) Now 1 drop of solution was added in slide and it was then covered with
cover slip.
6) Slide was then analyzed under light microscope.
Observation:
Fig11 Microscopic view of Industrial effluent:
No of bacteria present in sample
36
Protozoan cyst colony Entamoeba coli cyst
non pathogenic cyst of Protozoa
Macrophage cells cyst of gardia
37
Spiral fibres
Fig.12 Microscopic view of Waste water:
PROTOZOAN CYST COLONY Entamoeba histolitica
SPIRULLUM No. of bacteria present in sample
38
PORIFERA species Feacal matter
MOTILE TROPHOZOID OF PROTOZOA PORIFERA species
39
Results and Discussions:
PARAMETER INDUSTRIAL
EFFLUENT
INDUSTRIA
L
EFFLUENT
STANDARD
WASTE WATER WASTE
WATER
STANDA
RD
I. Physical Analysis of Water
1) Temperature of
water
31oC 30-35
oC 30
oC <40
oC
2) Conductivity of
water
2.56 mS 2 m S 2.7 mS 2 mS
II. Chemical Analysis of Water
1) pH in the water 6.8 5.5-9 7.21 6.5-8
2) Chloride in water 6.52 mg/ml 1 mg/ml 8 mg/ml 1 mg/ml
3) Acidity of water 205 mg/l - 125 mg/l -
4) Alkalinity of
water
420 mg/l <120 630 mg/l 500
5) TDS of water 500 mg/l <500 700 mg/l <500
III. Organic Constituents in Water
1) Free CO2 74.8 mg/l 22 48.4 mg/l 22
2) DO - -
3) BOD - -
4) COD 0.016 mg/ml - 0.008 mg/ml -
IV. Bacteriological Analysis of Water
1) MPN Test >= 1600
Coliforms
bacteria are
present in water
sample.
<400 34
Coliforms bacteria
are present in
water sample.
-
V. Parasitological analysis of water
non pathogenic
cyst of
Protozoa,
Protozoan cyst
colony, Bacilli,
Macrophage
cells
Entamoeba
histolitica, SPIRULLUM,
PROTOZOAN
CYST COLONY,
MOTILE
TROPHOZOID
OF PROTOZOA
Table 3 the analysis result and standard parameters of both Industrial effluent water sample and
Waste water sample are tabulated in below mentioned table
40
Results of water quality index obtained revealed many remarkable features
regarding the pollution status of Amanishah nala. Comparison between Physical,
Chemical, Bacteriological parameters and their respective standards shows many
differences in many parameters. Also by analyzing Parasitological parameters we can see
many species of parasites including protozoa, porifera and a huge number of bacterial
species. As we can see that conductivity, TDS, free CO2, free Cl2 and Alkalinity of both
Industrial effluent water sample and also the waste water sample are higher than their
respective standards. There are some parameters which lie under the limits of waste water
and industrial effluent samples, even during performing DO and BOD proper reading
could not been obtained i.e. free oxygen was nearly absent in both water samples,
possibility of which may be is because of growth of lots of micro organisms which are
facultative or obligate anaerobic.
All these results show negative characteristics of water i.e. contamination,
microorganisms, absence of free O2, free CO2 etc. Problem arises when this water is
taken in use for irrigation. The effluents are usually treated by physio-chemical treatment
followed by biological treatment process. However, such treatment systems are not
effective for removal of color, dissolved solids, trace metals, etc. and the effluents are
directly discharged into drains, public sewers, rivers, etc which ultimately become the
reason of high degree of pollution.
41
CONCLUSSION
These parameters definitely show that Amanishah nala is highly contaminated as
also stated by Dinesh Kumar et al (2005), Bhatnagar et al. (2006), Nupur Mathur,
Pradeep Bhatnagarit(2007). Although there are some parameters which come under the
tolerable range of the standards provided yet it is very important to be noticed that these
parameters, as shown above are for Industrial effluents and Waste water, not for
irrigation purpose but Amanishah nala water from a long time is being used for Printing,
Dying and most importantly for irrigation, which makes this water channel highly
polluted.
Overall findings indicated that wastewaters of the major industrial areas of Jaipur
city were not found good and should not be used for irrigation without prior treatment.
Immediate action could be taken by the people at household level like they should try and
find some other water sources for irrigation, filter plats should be installed in personal
level etc. as environmental and engineering measures at community level would take a
long time to be applied.
42
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Singh Vijendra and Singh C., Journal of Environ. Science & Engineering VOL. 48,No. 2,
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Websites:
http://www.pcd.go.th/info_serv/en_reg_std_water.html
http://academic.pgcc.edu/~kroberts/Lecture/Chapter%206/06-T06a_MPN-
Table_T.jpg
http://academic.pgcc.edu/~kroberts/Lecture/Chapter%206/06-T06b_MPN-
Table_T.jpg
www.jeb.co.in/journal_issues/200701_jan07/paper_18.pdf
http://www.water-treatment.com.cn/resources/discharge-standards/mauritius.htm
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