Physio chemical, microbiological and parasitological analysis of amanishah nala water from jaipur

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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

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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

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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

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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.

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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

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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

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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

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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.

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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

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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.

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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

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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.

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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.

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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.

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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.

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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

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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.

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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.

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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

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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

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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

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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

REFERENCES

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pollution of Amanishah Nallah by effluents of local textile industries in Sanganer,

jaipur(Rajasthan), P20-21 (2003)

Khan T.I., Kaur N. and Vyas P.C., Effects of industrial effluents on physicochemical

characteristics of Amani Shah Nallah-A case study. J. Env. Poll., 2(3),147-150(1995).

Singh Vijendra and Singh C., Journal of Environ. Science & Engineering VOL. 48,No. 2,

P. 103-108)Analytical Study of Heavy Metals of Industrial Effluents at Jaipur, Rajasthan

(India), (April 2006)

Singh Vijendra, Singh Chandel CP, Water quality of groundwater and wastewater of

Jaipur city for irrigation purpose. Aquacult, 6(1) (2005)

Kumar Dinesh, Jain Mukta, Dhindsa SS, Devanda HS, Singh, Physico-chemical

characteristics of Amanishah Nallah and neighboring ground water sources in Sanganer,

Jaipur. (2005)

Esabela, Sharma K. and Chauhan S.:Physico-Chemical profile of untreated irrigation

water from Amanishah nalla, Sanganer(Jaipur)(1998)

Sharma S. K., ‘Ground water pollution of Sanganer block of Jaipur district in

Rajasthan’,

Environment and Ecology, P 934-940(2004)

Sharma JD, Jain P, Sohu D, Quality status of groundwater of Sanganer tehsil in

Jaipur district. Nature Env. Polln. Techno, P 207-212,(2005)

Mathur Nupur and Bhatnagar Pradeep, Mutagenicity assessment of textile dyes from

Sanganer (Rajasthan), P- 1(2005)

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K.Verma Avnish, Prakash Ved and Saksena D.N., DrinkingWater Quality of Delhi, NCR

and Some Areas of Uttar Pradesh in India, p 101, (2005)

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three different places of Sivakasi 28(1) 105-108 (2007)

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Water Samples from Manipur River System, India, Vol. 14 (4) 85 – 89 (2010)

Salisu Dan’azumi, Mustapha Hassan Bichi, Industrial Pollution and Implication on

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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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Environmental sciences- A systematic approach, Dr. Rajni Johar Chhatwal

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