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



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  • 1. INTRODUCTION Water is an essential natural resource for sustaining life and our environment on thisearth. Water is always available in abundance on this planet. Water is not only vital to life but itis also a vital component of healthy functioning of any ecosystem (Simmons, 1999) as it is incontinuous interaction with the surrounding air and land and living things. Water is alsogeologically important because of its role in weathering, erosion, transportation and deposition ofsediment (The Atlas of Canada, 2004). Unfortunately, unless we use our water wisely, thewater bodies such as rivers, lakes, and groundwater etc. can become depleted or polluted, andunavailable or unsuitable for life. Water is not only the survival resource of all living beings butalso the main vector for all development activities and is integratedly related with all ecologicaland 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 anthropogenicactivities. The quality of water has undergone a change to an extent that the use of such watercould be hazardous. Increase in overall salinity of the ground water and/or presence of highconcentrations of fluoride, nitrate, iron, arsenic, total hardness and few toxic metal ions havebeen 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 theRajasthan State is also popularly known as Pink city and is situated towards central part of thedistrict. Jaipur is very much on the world tourist map, known for gem & jewelry and is alsopopular for Sanganer & Bagru prints. In the present study Sanganer town is selected as studyarea. Sanganer town situated 20 km away from the main city of Jaipur is located south of Jaipurand 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 byconstruction of Jhalaras (a big public well) in each of the nine Chowkaris (squares). In thebeginning of the 18th century the citizens used to draw water from 100 open wells and baorislocated in the city. The first water supply scheme for City Palace where water from open wellswas drawn with oxen and was supplied through a small canal. In the mid of 18th century the 1
  • 2. walled city was supplied water from Amanishah Nala through canals up to Chhoti Chopar andBari Chopar from where people used to take water. Piped-water supply to Jaipur for public beganin 1874 with the construction of a Reservoir of 4.5 MLD capacity across Amanisha Nala. Openwells were constructed in the Nala to meet the drinking water demand. Later on the Ramgarhdam was constructed in the year 1906. Due to increased population a scheme was taken up toaugment water supply from the lake and closed pipe lines were constructed for supply to thewhole city. Amanishah nala was then declared as a waste water channel. But due to varioususages of its water i.e. printing, dying, irrigation, which in a huge amount happens inSANGANER 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, directaniline black, which also includes many metal based dyes used for fastening colors. There arearound 700 varieties of dyes and dye intermediates produced in India, mainly direct dyes, aciddyes, reactive dyes and pigments. Most of these dyes have not been characterized regarding theirchemical 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 2
  • 3. Printing involves use of large amount of water and thus large quantity of waste water isalso generated. The untreated sewage water and waste water from textile industries (whichcontains 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, forgrowing vegetables and other crop plants. It certainly makes the part of food chain. The wastesreleased from such industries cause soil, surface and ground water pollution, besides causing anumber of adverse effects on agricultural products, animals and health of people living in thatarea as human beings in and around Sanganer in Jaipur district consume these vegetables andproducts 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 majorenvironmental threat because of the large amounts of water and dyes involved in themanufacturing process. Large amounts of chemically different dyes are employed for variousindustrial applications including textile dyeing. Dye production in India is estimated to be around64,000 tones, which is about 6.6% of the world production. The textile industry in India aloneconsumes up to 80% of the total dyestuffs produced. In Rajasthan state particularly, textile millsrepresent an important economic sector. Sanganer is famous for dyeing and printing of colorfuldresses, bed sheets, curtains, dress material and variety of other textiles. Most of the industries have been using many dyes as mentioned but the agony despitethe 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 seenaccumulated in many areas in absence of proper drainage Bulk of the textile products of theseindustries is exported. It is located about 15 km south of Jaipur, the State capital that has a 3
  • 4. population of more than two million people. The total area of Sanganer is about 635.5 Sq. km outof which, 12.9 Sq. km comprises the urban area. Most of the textile industries of Sanganer areconcentrated in this urban area. There are estimated to be around 500 block and screen printingunits in Sanganer. Among the total dyestuff consumption, it has been reported that textileindustry accounts for 67% of the total dyestuff market. These activities often lead to alteration ofwater quality by raising the physio-chemical parameters above the allowable limits andultimately results into Environmental pollution. Environmental pollution is one of the most horrible crises that we are facing today. Dueto the increased urbanization and industrialization surface water pollution has become an crucialproblem. It is necessary to obtain precise and appropriate information to observe the quality ofany water resources and the development of some useful tools to keep watch on the quality ofsuch 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 waterpollution but have also caused ground water pollution in the Jaipur city resulting in adverseeffects on the health. Voluminous liquid wastes are generated by the dyeing and printing industryand are disposed off in carrier channels (canals). In addition, some industrial units are alsopouring their effluents into the Amanishah Nala. These liquid wastes are also being used forirrigation purposes. The unused part of effluent water is allowed to accumulate near the bunds inthe peripheral areas giving adequate time period to this effluent water to percolate and reach thesaturated zone. Thereby degrading and deteriorating ground water quality. The polluted surfacewater flows as per hydraulic gradient and gets concentrated at favorable geomorphic locationswhere its flow is sluggish. There is an urgent warning and calls for measures to tackle the qualityhazards of ground water. It is therefore recommended that the liquid effluents should be treatedand beneficiated to remove the hazardous constituents before their disposal and also toencourage / motivate to use vegetable dyes. Alternatively, the dyes having higher concentrationof fluoride should be replaced by alternative dyes. It is also recommended to develop a propersewerage network system in urban agglomerate areas particularly in the walled city so as toprevent mixing with ground water. 4
  • 5. Parameters Reason for the analysisPhysical ParametersTemperature 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. o In water above 30 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 ParameterspH 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 5
  • 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 constituentsDissolved 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 is a measure of organic pollution to both waste andBOD 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 is an indicator of organics in the water, usually used inCOD 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 6
  • 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.MicrobiologicalTotal Coliform Count Microbiological test is to detect the Level of pollutions caused by living thing especially human who live or work in the areaFaecal Coliform Count 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. 7
  • 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. 8
  • 9. REVIEW OF LITERATURE Systematic Hydro geological Survey in the entire district was completed byGround Water Wing of Geological Survey of India & Central Ground Water Boardwhich 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 Programmesand five Water Management Training Programmes have been conducted. World WaterDay is also often celebrated in Jaipur. Literature on studies on the impact of waste water on agricultural crops reveal thatcrop plants and vegetables grown in the agricultural fields by using untreated waste waterwere adversely affected both qualitatively and quantitatively.Ganeshan and Manoharan (1983) studied the effect of cadmium and mercury ongermination of seeds, growth of seedling and matter production of Abelmoschus esculentsand found that cadmium is more toxic than mercury and the water of high concentrationof 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 surroundingpopulations and measured the levels of various physical and chemical parameters.Brown and Wilkins (1986), Dayama (1987) studied influence of dyeing and textilewaste water on nodulation and germination of Cicer aeritium.Rana and Masood (2002) conducted a pot study to investigate the toxic effects of certainheavy metals on the plant growth and grain yield of wheat (Triticum aestivum L.).S. K. Sharma, (2004) evaluated groundwater samples from Jaipur, Rajasthan, India todetermine their suitability for irrigation and drinking purposes. He found that the qualityof 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 9
  • 10. characteristic of the samples could be classified under moderate category and were goodfor household, irrigation and commercial purposes.BK Nayak, BC Acharya, UC Panda et al (2004) studied water quality parameters forthe entire Chilka lake covering a maximum of 23 sampling stations and found that pH ofwater was alkaline throughout the lake and both pH and salinity varied widely. HigherpH with low salinity zones reflected disintegration of submerged weeds. Correlationanalysis supported the increase of pH, high photosynthetic activity, high nutrients as wellas 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 wasindicative of increased quantity of industrial effluents. The reduced DO content was dueto hot ash slurry from thermal power plant. They also collected data from present studywhich reveals that there is is an inverse relationship between DO and BOD andportability of Irai river water is below the standard permissible limit.Moti R Sharma, AB Gupta, JK Bassin (2004) stated that the dissolved oxygen in thestream is below 4mg/l in a stretch of 2600m and therefore water is not fit for publicsupply, bathing, fish culture and wildlife in Hathli stream Shivalik Himalayan.Vijendra Singh, Chandel Singh (2005) analyzed groundwater and wastewater samplesfrom ‘Amanishah Nala’ and hand pump of seven industrial areas and adjacent localitiesof Jaipur city with the help of standard methods of APHA and Black. The values obtainedwere compared with standards of ISR, ICMR and WHO. The concentrations of variousparameters are within permissible limits in both groundwater and wastewater but definitecontaminations with special reference to EC, TDS and COD in wastewater have beenobserved which calls for at least primary treatment of wastewater before being used forirrigation. 10
  • 11. Dinesh Kumar et al (2005) monitored Sanganer nala and surrounding tube wells andstated that discharge of untreated industrial effluents and sewage in to nala havecontributed considerable pollution in the ground water in its vicinal areas, and is harmfulfor use in agriculture and drinking purposesJD Sharma, P Jain, D Sohu (2005) revealed that pH, EC and alkalinity of all thesamples from villages of Sanganer, Jaipur were very high which can be correlated withhigh TDS and chloride. Twenty eight percent villages contained high fluorideconcentration than permissible limit i.e., 1.5ppm. A positive correlation was observedbetween pH and fluoride, TDS and EC. Hardness showed negative correlation withfluoride and pH.SS Asadi SS, Padmaja Vuppala, M Anji Reddy (2005) stated that high concentrationsof total dissolved solids, nitrates, fluorides and total hardness were observed in fewindustrial and densely populated areas in Hyderabad indicating deteriorated water qualitywhile the other areas exhibited moderate to good water quality.Bhaskar Bhadra et al (2005) found higher range of alkalinity, ammonia content andchloride content in Torsa than Kaljani. River Kaljani showed higher COD range thanTorsa. Mean BOD value of both these rivers ranged between 0.93-1.65 mg/l. OverallTDS content of Kaljani was found to be lower than Torsa. Maximum phosphate contentwas observed at the downstream of both the rivers.Ram Chandra, RN Prasad (2005) studied various water quality parameters related tothe deterioration in water quality of Surya Kund, Lohargal (Rajasthan) during the massbathing of religious importance and said that it is necessary to take adequateprecautionary measures to prevent outbreak of any epidemics.R Rajaram, M Srinivasan, M Rajasegar (2005) said that nutrient concentrations werehigher during monsoon season and low during summer season at two stations of Uppanarestuary, Cuddalore in relation to effluent discharges from SIPCOT industries. There are 11
  • 12. 44 industries discharging their effluents into Uppanar estuary, which may influence thebiota.SR Vishnoi, PN Srivastava (2005) collected water sample from three different sites ofriver Jojari at Salawas, Jodhpur and carried hydro biological studies. They found thatthe pH, chloride, salinity, total alkalinity, total hardness, dissolved oxygen and TDS wereabsolutely higher than the standard values of portable water on account of contaminationof river due to industrial effluents. The river has become unsuitable for the growth andsurvivability of aquatic flora and fauna. The pollution impact was found to bepredominant during summer and minimal during monsoon season.Bhatnagar et al. (2006) reported that waste water effluents from textile dyeing andprinting industries of Sanganer were discharged directly, without any treatment, intoAmanishah nala drainage. The drainage water takes the dissolved toxicants to flora andfauna, including crops and seasonal vegetables, being grown in the land adjoining thenala drainage. The mutagenic potential of vegetables irrigated by water of Amanishahnala drainage was investigated by them.Marques et al. (2007) studied the levels of zinc accumulated in field conditions, byroots, stems, and leaves of Rubusulmifolius and Phragmites australis, the speciesindigenous to the banks of a stream in a Portuguese contaminated site.Oporto et al. (2007) studied elevated Cd concentration in potato tubers due to irrigationwith river water contamination by mining in Potsi, Bolivia.Nupur Mathur, Pradeep Bhatnagarit(2007) collected water samples from theagricultural fields of Trigonella foenumgraecum grown in Sanganer area. These sampleswere found to contain pH in the range from 8.4 to 11.9. Electric conductivity ranged from0.564 to 3.203mmhos/cm, whereas concentrations of total dissolved solids were found inthe range from 750 to 4670 mg/L and Chlorides from 315.40 to 1304.60 mg/Lrespectively. The data indicate that effluent concentrations in the waste water ofAmanishah nala have considerably been increased during the past few decades. 12
  • 13. Sikka et al. (2008) stated that the disposal of industrial and sewage water was a problemof increasing importance throughout the world. In India and most of the developingcountries untreated sewage and industrial wastes are discharged on land or into therunning water streams which is used for irrigating crops.The objectives of these scientific investigations has been to determine the agro-chemistryof the water and to classify the waste water in order to evaluate the effect of pollutants onplants, and the water suitability for irrigation uses and the present report is also aimed atit. The analysis of waste water is made at pre-flowering, flowering and post floweringstages in order to make comparison of the quality of water at these stage sand in turncorrelate with external, internal morphological changes, if any, at the three stages ofgrowth. 13
  • 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 sample1. 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. 14
  • 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. 15
  • 16. Theory: NaCl is the main substance responsible for chloride conc. in water. Chloride isestimated on the basis of argentometric method in high chloride is precipitated withpotassium chromate.Permissible limit for chloride is 250 mg.AgNO3 + Cl- AgCl (white ppt.) + NO3-2 AgNO3 + K2CrO4 Ag2CrO4(Brick Red) + 2KNO3Procedure: 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 20Calculation: 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 16
  • 17. EXPERIMENT 3Objective: To determine the acidity of the given water sample.Reagents: NaOH (0.05 N) dissolve 1gm in 500 ml DW, Methyl orange indicator,Phenolphthalein indicatorTheory: Acidity of water can be neutralized by strong base. In natural unpolluted freshwater, 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, usingmethyl orange and phenolphthalein as indicators. If sample has strong mineral acid andits salt just titrates to pH 3.7, using methyl orange as an indicator then it is called methylorange acidity.If sample is titrated to pH 8.3 then phenolphthalein indicator should be used. Theresulting value is called total acidity.Result of acidity should always be mentioned with pH titration method is suitable mainlyfor 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. 17
  • 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 4Objective: To estimate the total alkalinity (carbonate and bicarbonate) in the given watersample.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 inwater 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 incarbonate, bicarbonate with little concentration of other alkalinity imparting ions. Total alkalinity, carbonates, bicarbonates can be estimated by filtering the samplewith strong acid either HCL or H2SO4. First it is titrated to pH 8.3 using phenolphthalein as an indicator then further pHbetween 4.2- 5.4 with methyl orange indicator. In first, the value is calledphenolphthalein alkalinity and in second is total alkalinity. Value of carbonate,bicarbonate and hydroxyl ion can completed from these two types of alkalinities. 18
  • 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 19
  • 20. EXPERIMENT 5Objective: 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 largelygovern the physico- chemical properties of water and in turn have an indirect effect onthe flora and fauna. Total dissolved solids (TDS) are determined as the residue left afterthe 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 20
  • 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 21
  • 22. EXPERIMENT 2Objective: Determination of dissolved oxygen (DO) of water.Requirements: Water sample (tap water), Sodium thiosulphate (0.025 N), (dissolve 1.24gm 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 distilledwater), Starch indicator (1%), concentrated sulphuric acid, DO bottles.Theory:Dissolved oxygen of water is of paramount importance to all living organisms. Thepresence 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 underthe influence of temperature, salinity, water movements etc. whereas the later is abiological process and depends on availability of light and the rate of metabolic processesresulting in diurnal fluctuations.The estimation of DO is done by titrimetric method. The oxygen of the water combineswith manganous hydroxide, which on acidification liberates iodine equivalent to that ofoxygen fixed. This iodine is titrated by standard sodium thiosulphate solution usingstarch 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. 22
  • 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. 23
  • 24. EXPERIMENT 3Objective: 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 asmeasured by the volume of oxygen required by bacteria to metabolize it under aerobicconditions. It is good index of the organic pollution. If the amount of organic matter insewage is more, the more oxygen will be utilized by bacteria to degrade it. Dumpingsewage that contains high BOD increases the concentration of soluble organiccompounds in the aquatic body where it is discharged. Digestion of these organiccompounds in natural ecosystems, such as lakes, rivers, can deplete available O2 andresulting in asphyxiation of fish.The BOD of a water sample is generally measured by incubating the sample at 20degrees for five days in the dark under aerobic conditions. In tropical and sub-tropicalbelts, where the temperature and rate of metabolic activities are higher, the incubationshould 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) 24
  • 25. Calculations: BOD (mg/l) = D1 – D2 Where, D1 = Initial DO of sample. D2 = DO after 3 days incubation. 25
  • 26. EXPERIMENT 4Objective: Determination of chemical oxygen demand (COD) of water sample.Requirement: Water sample, potassium dichromate solution (0.1 N), sodiumthiosulphate (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 ofvarious chemically oxidizable organic substances of different nature entering the aquaticsystems, BOD alone doesn’t give a clear picture of the organic matter content of thewater sample.In addition, the various toxicants in the sample may severely affect the validity of theBOD 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 oxygenconsumed 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 chemicaloxidant such as potassium permanganate or potassium dichromate. In the permanganatemethod, the organic matter is first oxidized with a known volume of KMnO4 and thenexcess of oxygen is allowed to react with potassium iodide to liberate iodine in amountsequal to the excess oxygen, which is estimated titrimetrically with sodium thiosulphatesolution 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. 26
  • 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 4Calculation: 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 27
  • 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. 28
  • 29. PRESUMPTIVE COLIFORMS TEST:The presumptive Coliforms test is used to detect coliforms in a water sample. In this testlactose fermentation tubes are inoculated with different water volumes and production ofacid and gas from the fermentation of lactose in any of the tubes is a presumptiveevidence of coliforms in the water sample. The lactose broth used in this test is selective for the isolation of coliformsbecause of the addition of the bile and lauryl sulphate or brilliant green. A pH indicatorsuch as Bromocresol purple is also added to lactose broth for the detection of acid. Thecolor of the indicator changes to yellow with the production of acid from lactose.Requirements: Water sample (100 ml), Lactose broth medium, Durham tubes (15), 10ml double strength lactose broth tubes (LB2X) (5), 5 ml single strength lactose brothtubes (LB1X) (10), Sterile pipettes, one each of 10 ml, 1 ml and 0.1 ml capacity, Bunsenburner, 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. 29
  • 30. 10) Incubate all the 15 inoculated tubes aerobically at 35 degree centigrade for 48 hours.Observations:Fig.6 Lactose broth tubes with positive testAll 15 tubes +ve in both industrial effluent and waste water 30
  • 31. CONFIRMED COLIFORMS TESTThis test is used to confirm the presence of coliforms and determine the MPN value inwater sample showing positive or doubtful presumptive test. In the confirmed test, watersamples from all the positive presumptive lactose broth tubes are inoculated into tubes ofbrilliant 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 toestimate the population of coliforms, which means that the result obtained in expressed asthe most probable number (MPN) of coliforms. A count of number of lactosefermentation tubes/brilliant green lactose bile broth showing production of gas followingthe incubation period is taken and MPN is found by matching the results with thoseprovided in the statistical table.Requirements: 10 ml brilliant green lactose bile broth fermentation tubes (the numberdepending upon the tubes showing positive presumptive test), Durham tubes, Inoculationloop, 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 31
  • 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 32
  • 33. COMPLETE COLIFORMS TESTCompleted test is used to establish the presence of Coliform bacteria and as confirmatorytest for the presence of E.coli in a water sample. In the completed test, the samples fromthe positive brilliant green lactose bile broth from the confirmed test are streaked onto aselective differential medium for coliforms and inoculated into lactose broth tube as wellas streaked on a nutrient agar plate to perform Gram staining. The medium commonlyused is eosin-methylene blue (EMB) that is selective in nature because of the dyemethylene blue which inhibits the growth of Gram-positive bacteria, allowing the growthof Gram-negative bacteria EMB is differential in nature in that lactose fermentingbacteria gives colored colonies (a positive confirmed test) due to the formation of acomplex in EMB that precipitates out onto the coliforms colonies. Non-lactosefermenters produce colorless colonies on EMB agar. If there is production of acid and gasin the inoculated lactose broth and there are rod shaped bacteria showing Gram-negativereaction, these confirm the presence of E.coli in the water sample and are considered apositive completed test.Requirements: EMB agar plates, 24 hours coliforms confirm positive brilliant greenlactose bile broth culture (from the confirmed test), 5 ml brilliant green lactose brothfermentation 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 inpositive samples.Results: Coliforms bacteria are present in water sample. Hence the water sample is notpotable.Fig.9 Bacterial culture with Industrial effluent: 33
  • 34. Fig.9 10 ml 1 ml sample sample 0.1 ml sampleFig.10 Bacterial culture with Waste water: 10 ml sample 1 ml sample 34
  • 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 35
  • 36. Protozoan cyst colony Entamoeba coli cystnon pathogenic cyst of Protozoa Macrophage cells cyst of gardia 36
  • 37. Spiral fibresFig.12 Microscopic view of Waste water:PROTOZOAN CYST COLONY Entamoeba histolitica SPIRULLUM No. of bacteria present in sample 37
  • 38. PORIFERA species Feacal matterMOTILE TROPHOZOID OF PROTOZOA PORIFERA species 38
  • 39. Results and Discussions: PARAMETER INDUSTRIAL INDUSTRIA WASTE WATER WASTE EFFLUENT L WATER EFFLUENT STANDA STANDARD RD I. Physical Analysis of Water 1) Temperature of 31oC 30-35 oC 30oC <40 oC water 2) Conductivity of 2.56 mS 2mS 2.7 mS 2 mS waterII. 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 420 mg/l <120 630 mg/l 500 water 5) TDS of water 500 mg/l <500 700 mg/l <500III. 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 <400 34 - Coliforms Coliforms bacteria bacteria are are present in present in water water sample. sample.V. Parasitological analysis of water non pathogenic Entamoeba cyst of histolitica, Protozoa, SPIRULLUM, Protozoan cyst PROTOZOAN colony, Bacilli, CYST COLONY, Macrophage MOTILE cells 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 39
  • 40. Results of water quality index obtained revealed many remarkable featuresregarding the pollution status of Amanishah nala. Comparison between Physical,Chemical, Bacteriological parameters and their respective standards shows manydifferences in many parameters. Also by analyzing Parasitological parameters we can seemany species of parasites including protozoa, porifera and a huge number of bacterialspecies. As we can see that conductivity, TDS, free CO2, free Cl2 and Alkalinity of bothIndustrial effluent water sample and also the waste water sample are higher than theirrespective standards. There are some parameters which lie under the limits of waste waterand industrial effluent samples, even during performing DO and BOD proper readingcould 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 arefacultative 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 istaken in use for irrigation. The effluents are usually treated by physio-chemical treatmentfollowed by biological treatment process. However, such treatment systems are noteffective for removal of color, dissolved solids, trace metals, etc. and the effluents aredirectly discharged into drains, public sewers, rivers, etc which ultimately become thereason of high degree of pollution. 40
  • 41. CONCLUSSION These parameters definitely show that Amanishah nala is highly contaminated asalso stated by Dinesh Kumar et al (2005), Bhatnagar et al. (2006), Nupur Mathur,Pradeep Bhatnagarit(2007). Although there are some parameters which come under thetolerable range of the standards provided yet it is very important to be noticed that theseparameters, as shown above are for Industrial effluents and Waste water, not forirrigation 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 highlypolluted. Overall findings indicated that wastewaters of the major industrial areas of Jaipurcity 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 andfind some other water sources for irrigation, filter plats should be installed in personallevel etc. as environmental and engineering measures at community level would take along time to be applied. 41
  • 42. REFERENCESYadavi K., Sharma Smita, Saint Yashoda, Khan T., and Sharma Shweta: Study onpollution 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 physicochemicalcharacteristics 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 ofJaipur city for irrigation purpose. Aquacult, 6(1) (2005)Kumar Dinesh, Jain Mukta, Dhindsa SS, Devanda HS, Singh, Physico-chemicalcharacteristics of Amanishah Nallah and neighboring ground water sources in Sanganer,Jaipur. (2005)Esabela, Sharma K. and Chauhan S.:Physico-Chemical profile of untreated irrigationwater from Amanishah nalla, Sanganer(Jaipur)(1998)Sharma S. K., ‘Ground water pollution of Sanganer block of Jaipur district inRajasthan’,Environment and Ecology, P 934-940(2004)Sharma JD, Jain P, Sohu D, Quality status of groundwater of Sanganer tehsil inJaipur district. Nature Env. Polln. Techno, P 207-212,(2005)Mathur Nupur and Bhatnagar Pradeep, Mutagenicity assessment of textile dyes fromSanganer (Rajasthan), P- 1(2005)Yasser Abdul Kadar Al-Gahwari, Physico-Chemical parameters and microorganisms aswater qualityindicators of TELUK BAHANG RESERVOIR AND BATU FERRINGHITREATMENT PLANT, P 68-69(2007) 42
  • 43. 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) R. Radha , Dharmara K. j and Kumari Ranjitha , A comparative study on the physicochemical and bacterial analysis of drinking,borewell and sewage water in the three different places of Sivakasi 28(1) 105-108 (2007) SINGH M. R, GUPTA ASHA, BEETESWARI, KH., Physico-chemical Properties of Water Samples from Manipur River System, India, Vol. 14 (4) 85 – 89 (2010) Salisu Dan’azumi, Mustapha Hassan Bichi, Industrial Pollution and Implication on Source of Water Supply in Kano, Nigeria, P 101(2010) 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 Bibliography: Environmental sciences- A systematic approach, Dr. Rajni Johar Chhatwal Environmental science and Biotechnology, A G Murugesan and C Rajakumari Environmental Biotechnology- Basic concepts and Applications. Environmental science, Rintu Banerjee 43