+E=.06. cE. ake © Experiment No. 10 etermination of Biochemical Oxygen Demand’ _ STANDARD PHOTOSTATE ue KUET MAINGATE RODUC When biodegradable organic matter/waste (the most common category of pollutant affecting surface water) is released into a water body, microorganisms (especially bacteria) feed on the wastes, breaking it down to simpler organic and inorganic substances. When this decomposition takes place in an acrobic environment (i.c., in the presence of oxygen, Eq. 1), it produces non-objectionable, stable end products (¢.g., CO;, S04, PO,, and NO,) and in the process draws down the dissolved oxygen (DO) content of water. « Organic matter +O, = CO, + H,O + Newcells + Stable products (1) s (bacteria), When insufficient oxygen is available or when oxygen is exhausted by the aerobic decomposition of wastes, different set of microorganisms carry out the decomposition anaerobically (Eq. 2) producing highly objectionable products including H,S, NH,, and CH, a Organic matter. = CO, + CH, + Newcells + Unstable products Q) (bacteria) The amount of oxygen required by micro-organisms to oxidize organic wastes acrobically ts called biochemical oxygen demand (BOD). BOD may have various units, but most often it is expressed in mg of oxygen required per liter of water/wastewater (mg/L). The total amount of oxygen that will be required for bio-degradation of is an important measure of the impact that a given waste stream would have on the receiving water body. Dissolved oxygen is the most commonly used indictor of the general health of a surface water body. If DO goes below 4 to 5 mp/L (due of decomposition o1 organic wastes), forms of life that can survive Begin to be reduced, When anacrobic condition exists, higher life forms are killed or driven off, Noxious condition, including floating sludge, bubbling, odorous gases, slimy fungus wth prevails AV1SOLOHd GUVGNV1S ORY The biochemical oxygen demand (BOD) determination is an empirical test in which standardized laboratory procedures are uscd to determine the oxygen requirements of wastewater, effluents and polluted waters. It has become a standard practice to simply measure and report the oxygen demand over a 5-day period, realizing that the ultimate oxygen demand (for complete degradation of organic matter) is considerably higher (and would take a much longer time to determine in the laboratory) The 5-day BOD oF BOD, is the amount of oxygen consumed by micro-organisms during the first 5 days of biodegradation. In its simplest form, a BOD, test would involve putting a sample of waster/wastewater into a stoppered bottle, measuring the dissolved oxygen (DO) of the sample at the beginning of the test and again at the end of five days. The difference in DO would be the BOD, of the water/wastewater. Light must be kept out of the bottle to keep ‘ah 1 adding oxygen by photosyr*h~ is and the stopper is used to keep air from nw:replenishing DO from outside, To standardize the procedure, the test is run at a fixed Bs ‘temperature of 20 °C. Since the oxygen demand of typical waste is sever hundred milligrain, per liter, and since the saturated value of DO for water at 20 °C is only 9.1 mg/L, it is usually necessary to dilute the sample to keen final DO above zero. If during the five days of experiment, the DO drops to zero, then the test 1s invalid since more oxygen would have been removed had more been available. The five-day BOD ofa diluted sample is given by BOD, = [Do, - Do) x DF. @) where, D.F.* ilution Factor = (Vol. of wastewater + dilution water) / (Vol. of wasteweater) In some cases, it becomes necessary to seed the dilution water with microorganisms to ensure _ ‘that there is an adequate ate bacterial pe population to carry out the biodegradation. In. such ¢ cases, two sets of BOD bottles must be prepared, one for just the seeded dilution water (called the_ “blank") and the other for the ¢ of wastewater and dilution water. The change in DO in both are measured. The oxygen demand of the wastewater (BOD.) is then determined from the following relationship: BOD, V, = BOD, V, + BOD, V, (4) : where, BOD,, is the BOD of the mixture of wastewater and dilution water and BOD, is the BOD of the dilution water alone,; V,, and V, are the volumes of wastewater and dilution water, respectively, in the mixture, and V,, = V, + Vy : lution wi bm A wide variety of dilution waters have been used for BOD work. Natural surface waters have been used, but they suffer from a number of disadvantages including variable BOD, variable ‘micro-organism population, and variable mineral content. Tap water has also been used, but it also suffers from many of the limitations of surface water plus the possibility of toxicity from chiorine residuals. Synthetic dilution water prepared by adding different salts to distilled water has been found to be the most ideal for use as dilution water. The pH of the dilution water should be between 6.5 to 8.5. Potassium, sodium, and magnesium salts are added to distilled water to give buffering capacity and proper osmotic pressure. Femc chloride, magnesium sulfate, and ammonium chloride are added to supply the requirements for iron, sulfur, and nitrogen. The phosphate buffer provides phosphorus that may be needed The_dilution_water_is_usually well_acrated_to_raise_its DO level before mixing with wastewate A wide variety of materials have been used for “seeding” purpose. Experience has shown that domestic wastewater, particularly from combined sewer systems, provides a well balanced population of mixed micro-organisms, ‘Usually 2 ml of wastewater per liter of diluted water is sufficient, Some river waters are also satisfactory, but care must be taken to avoid ui contain algae or nitrifying bacteria in significant amount. The dilution water should always be “seeded” to ensure a uniform population of micro-organisms.STANDARD PHOTOSTATE N i f WET i odeling of BOD: It is often assumed that the rate of decomposition of organic waste is proportional to the amount of waste available. _If L, represents the amount of oxygen demand left after time ¢, then, assuming a first order reaction, we can write, dL jdt = -kL, ) where, k = BOD reaction rate constant (time). Integrating, we get, © where, L, is the ultimate carbonaceous oxygen demand, which is the total amount of oxygen required by microorganisms to oxidize the carbonaccous portion of the waste. Now, L, = BOD, + L, q) and combining, we get, BOD, = L, (1 -e™) (8) Figure 1(a) and 1(b) shows the graphical representations of Eq, (6) and Eq. (8), respectively. Besides the oxidation of carbonaceous organic matter, a significant additional demand may be caused by the oxidation of nitrogenous compounds. To distinguish this nitrogenous biochemical oxygen demand from the carbonaceous biochemical oxygen demand, the two are sometimes referred to as NBOD and CBOD, respectively. Figure 2 illustrates the two demands for a typical municipal waste. Note that the NBOD does not begin to exert itself for at least 5 to 8 days, so most 5-day BOD tests are not affected by the nitrification process Theory of DO Determination: The reactions involved in the various steps of the Winkler Method dissolved oxygen (DO) determination are presented below: Mangunous sulfate reacts with potassium hydroxide in the alkaline potassium iodide solution o produce a white precipitate of manganous hydroxide. MnSO, + 2KOH' = Mn(OH), + K,SO, (9) Ifthe white precipitate is obtained, there was no dissolved oxygen in the sample and there is no need to proceed further. A brown precipitate shows that oxygen is present and reacted with the nianganous hydroxide. The brown precipitate is manganic basic oxide and formed as follows: 2Mn(OH), + O, = 2MnO(OH), (10) Upon the addition of (sulphuric) acid, this precipitate is dissolved, forming manganie sultate MnO(OH), + 2H.SO, = Mn(SO,); + 3H,0 an asebeh adrenalin A; 800, *cgit 26 (Oxveen somuimeg. 800) Tome Tome i “ ta Figure 1, Idealized carbonaceous oxygen demand: (a) the BOD remaini tine. and (Ih) the oxyge « consumed or the biochemical oxygen demand as a function of ume Oxy9en consumes Ore sy « $ 6 7 8 8 WN Wa 13 1 15 16 17 18 19 20 Tome (devs Figure 2. Illustration of Carbonaceous and Nitrogenous Biochemical Oxygen Demand.‘his compound immediately reacts with potassium iodide, liberating iodine and resulting 1n the typical iodine (bluc) coloration of the water. Mn(SO,), + 2KI = MnSO, + K,SO, + 1; (12) ied by these reactions is equivalent to the quantity of oxygen ‘ént_in the sample. The quantity of iodine is determined by titrating a portion of the if Sodium thiosulfate solution. 2Na,$,0, + 1, = Na,S,O, + 2Nal (13) 1. Manganous sulfate solution (57) . Alkaline potassium iodide solution (58) 0.025N sodium thiosulfate (58) . Starch solution (indicator) (60) Concentrated sulfuric acid. ween APPARATUS 1. BOD bottle nos. 2. Beaker (250 ml) 2 Uno. 3. Measuring cylinder : Tn. 4. Dropper 21 no. 5. Stirrer : 1 no. PROCEDURE (for determination of DO) Hill two BOD bottles with sample (or diluted sample); the boities should be completely filled. Determine initial DO (DO,) in one bottle immediately after filling with sample (or diluted sample). Keep the other bottle in dark at 20 °C and afer particular days (usually 5-days) determine DO (Do,) in the sample (or diluted sample). Dissolved oxygen (DO) is determined according to the following procedure: (1) Add 1 mil of manganous sulfate solution to the BOD bottle by means of pipette, dipping in end of the pipette just below the surface of the water. (2) Add | ml of alkaline potassium iodide solution to the BOD bottle in a similar manner. (3) Insert the stopper and mix by inverting the bottle several times. (4) Allow the “precipitates” to settle halfway and mix again. (5) Again allow the “precipitates” to settle halfway. (6) Add 1 ml. of concentrated sulfuric acid. Immediately insert the stopper and mix as before. (7) Allow the solution to stand at least 5 minutes. (8) Withdraw 100 ml of solution into an Erlenmeyer Mask and immediately add 0.025N sodium thiosulfate drop by drop from a burctte until the yellow color almost disappears (9) Add about Lm. of starch solution and continue the addition of the thiosulfate solution until the blue color just disappears. Record the ml. of thiosulfate solution used (disregard any retum of the blue color). weiCALCULATION Dissolved oxygen, DO (mg/l) = ml of 0.025N sodium thiosulfate added x 2 Calculate BOD of the sample according to Eq. 3 or Eq. 4. ASSIGNMENT QUESTIONS (1) A sample of sewage is mixed with water (no seeding done) in the ratio of 1:20 (ie., 1 ml of sewage diluted to 20 ml by adding water) for BOD test. The initial DO is 8.5 mg/L and final DO, after 5 days, is 3.1 mg/L. Calculate BOD, of the sewage (2) A test bottle containing just seeded dilution water has its DO level drop by 0.8 mg/L in a S-day test. A 300 ml BOD bottle filled with 30 ml of wastewater and the rest with seeded dilution water experiences a drop of 7.3 mg/L in the same period (5-day). Calculate the BOD, of the wastewater. (3) In a BOD test on a diluted wastewater sample (1:20 dilution, but not seeded), the initial DO is 8.4 mg/L and final DO afer 5 days is 4.2 mg/L. If the reaction rate constant is 0.22/day, calculate: (a) 5-day BOD of the wastewater, (b) Ultimate carbonaceous BOD of the wastewater, (c) Remaining Oxygen demand afler S-days. oe (ne og todulphele ud X 2) x lore iy 0: 025(N cease DO (ry/e) -43-a o4 @ ExpcrimentNo.1| @& Determination of Chemical Oxygen Demand: INTRODUCTION The chemical oxygen demand (COD) test is widely used as a means of méa8uring the organic strength of domestic and industrial wastes. ‘This test allows measurement of a waste in terms of the total quantity of oxygen required for oxidation to carbon dioxide and water. The test is | based on the fact that all organic compounds, with a few exception, can be oxidized by the action of strong oxidizing agents under acid conditions. During the determination of COD, organic matter is converted to carbon dioxide and water regardless of the biological assimilability of the substance. For example, glucose and lignin arc both oxidized completely. As a result, COD values are greater than BOD values, | especially when biologicall eae orgs matter (e-g., lignin) is present. #> One of the chief limitations of COD test is its inability to differentiate between biologically oxidizable and biologically inert organic matter. In addition, it does not provide any evidence of the rate at which the biologically active material would be stabilized under conditions that exist in nature. The_major advantage of COD test is the short time required for evaluation, The determination can be made in about 3 hours rather than the S-days required for the measurement of BOD. For this reason, it is uscd as a substitute for the BOD test in many linstances. i x Potassium dichromate or potassium permanganate is usually uscd as the oxidizing agent in The determination of COD. In this class potassium permanganate would be used in the determination of COD. Potassium permanganate is selective in the reaction and attacks the carbonaceous and not the nitrogenous matter. In any method of measuring COD, an excess of oxidizing agent must be present to ensure that all organic matter is oxidized as completely as possible within the power of the reagent. This fequires that @ reasonable excess be present in all samples, It is necessary, therefore, to measure the excess in some manner so that the actual amount can be determined. For this purpose, a solution of a reducing agent (e.g., ammonium oxalate) is usually used |. Dilute sulfuric acid (63) |2. Standard potassium permanganate (65) 3. Standard Ammonium Oxalate Prepared by Dr. M. Ashraf Ali, Associate Professor, Department of Civil Enginecring, BUET 49APPARATUS 1. Beaker (250 ml): 1 no, 2. Dropper :1no, 3. Stirrer = 1 no. PROCEDURE (For water) (1) Pipette 100 ml of the sample into a 250 ml Erlenmeyer flask. (2) Add 10 ml. of diluted sulfuric acid and 10 ml of standard potassium permanganate. (3) Heat the flask in a boiling water bath for exactly 30 minutes, keeping the water in the bath above the level of the solution in the flask. The heating enhances the rate of oxidation reaction in the flask. (4) If the solution becomes faintly colored, it means that most of the potassium permanganate have been utilized in the oxidation of organic matter. In such a case, repeat the above using a smaller sample diluted to 100 ml with distilled water. (5) After 30 minutes in the water bath, add 10 ml of standard ammonium oxalate-into the flask. This 10 ml ammonium oxalate, which is a reducing agent, is just equivalent to the 10 ml potassium permanganate (oxidizing agent) added earlier. The excess of reducing agent (aimonium oxalate) now remaining in the flask is just equivalent to the amount of” the oxidizing agent (potassium permanganatc) used in the oxidation of organic matter. (6) The quantity of ammonium oxalate remaining in the flask is now determined by titration with standard jum permanganate, Titrate the content of the flask, while hot, with standard potassium permanganate to the first pink coloration. Record the ml of potassium permanganate used. CALCULATION COD (mg/l) = [mL of KMaO, used in Step (6) x (100) / mL of sample used) A MENT (1) What are the principal advantages and disadvantages of the COD test over the BOD test. (2) Explain why COD values are usually greater than BOD values. (3) What could be inferred from the following analytical results conceming the relative ease of biodegradability of each waste? 5-dayBOD COD Waste (mg/L) (mg/L) A 240 300° B 100 500 c 120 24061 Experiment No. 12 Chemical Coagulation (Alum Coagulationy INTRODUCTION Surlice water generally contains a wide variety of colloidal impurities that may cause the alee to appear turbid and may impart color to the water, Colloidal particles that cause color and turbidity are difficult to separate from water because the particles will not settle by gravity and are so small that they pass through the pores of most common filtration media, In Onder to be removed, the individual colloids must aggregate and grow in size so that they can sctile by gravity. Chemical agents are used to promote colloid aggregation by destroying the forces that stabilize colloidal particles. The process of destroying the stabilizing forces and causing aggregation of colloids is referred 10 as chemical coagulation, Coagulation involves relustion of electrical forces of repulsion and promotion of “chemical type’ between colloids, which eventually results in settling of the colloids and a removal of turbidity and color. i c which may result in resuspension of the colloids. Hence optimum coagulant doses are deterimined through laboratory model tests where the water to be treated are subjected to a ‘of doses of a coagulant and the removal efficiencies are observed. ny authors use the term “cougudation™ to describe the process by which the charge on particles is destroyed, and the term “floceudation” to describe the aggregation of particles into larger units. The chemical used for this purpose on cougudants used in water and wastewater treatment are such os alum, ferric chloride and ferric sulfate. called are called cougulants. The most Juminum and ferric salts The common metal salt alum (aluminum sulfate) isa good coagulant for water contaming, ‘apne iable organic matter. ‘The chemical formula used for commercial alum_ is Al Son e410. Once dissolved in water, aluminum forms hydroxo-complexes and solids jee. OIDs), AKO), AKOTD:". AMOMDs; Egy. 1-5] and as a tesult pH of water is Tivcral especially if alkalinity of water is low. Theoretically, cach mg/l of alum consume “Inprevimately 0.50 mgll (as CaCOs) of atkatinily. For water with Tow alkalinity, this may “esalvin significant reduction in pFT That may interfere with formation of aluminum hydrovide flocs Ifthe alkalinity is insufficient, coagulam aids such as lime (CaQH;), soda ash (Na.CO}), activated silica and polyelectrolytes are used to provide the necessary alkalinity. tron coagulants operates over a wider pI range and are generally effective in removing wbadiy and color from water, However, they are usually more costly M(SO)). ISO (alum) “ZAI 3 S00 w AI © 3 HO = AKOLD() + 31 Q) Aly + 1hO = AKON)" + 1 3) AI + HO = AKOH):’ + 2H" (4) MY + HO = AWON)Y +H" (3) and color, coagulation with alum and eavy metal ¢.leasl arsenic) from water, In Besides removal of turbidity sultiue is also widely used for removal o Pegpsued by Des M. Ashrat Ali, Assogiate Professor, Departement of Civit Logineening, DUETthis process heavy metal ions are_primarily removed by adsorption (and subsequent recipitation) onto fed Mocs of metal (cither aluminum or tron) hydroxides. Coagulation with ssfully used for removal of arsenic from water. a REAGENTS (1) Standard Alum solution. (2) Standard Arsenic solution (if As removal is to be demonstrated) APPARATUS (1) Coagulation (stir (2) pH meter (3) Turbidity meter (4) Glass beakers (1000 ml; 6 nos) ing) device PROCEDURI (1) Determine pit and turbidity of the water to be treated. You may be instructed to determine color and arsenic concentration of the water to be treated (if removal efficiencies of these parameters are to be determined). (2) Fill six 1000 ml beakers each with 500 ml water to be treated. (3) Add required (as instructed by teacher) coagulant (standard alum solution) to each beaker. (4) Mix the samples in the beaker with the help of the stirring device, Subject the samples to one minute of rapid mixing followed by 15 minutes of slow mixing (about 40 rpm). (5) Allow the flocs to settle down for about 15 minutes. Observe the characteristics of the flocs and the settling rates, (6) Collect the supernatant from each beaker and measure pil and turbidity ofeach. You may be instructed to measure color and arsenic concentration of the supematant samples (if removal efficiencies of these parameters are to be determined). (7) Plot pH versus alum dose in a graph paper and observe effect of alum dose on pH. (8) Plot turbidity versus the coagulant (alum) dose ina graph paper. Determine optimum dose of alum from this plot. (9) Arsenic and color removal efficiencies could also be determined if measurements of color and arsenic concentrations are carried out iM. QUESTIONS (1) What do you understand by coagulation and flocculation 2 Which coagulants are most commonly used for water and wastewater treatinent (2) Why addition of alum may result in a drop in ptf value. Discuss the effect of alum dose on pH from your experimental results. : (3) What is the primary mechanism by which heavy metal ions are removed during coagulation:Experiment No. 13 Residual Chlorine and Chlorine Demand: Break Point Chlorination’ INTRODUCTION Chlorination of public water supplies and polluted waters serves primarily to destroy or de- activate disease-producing microorganism” ‘The practice of disinfection with chlorine is widely practised. Chlorination may pro’ » leetse effects including taste and odor problem. In recent years, chlorination has been found to produce trihalomethanes (THMs) and other organics uf health concem (THMs are suspected human carcinogens). Thus, use of alternative disinfectants, such as chlorine dioxide and ozone that do not cause this particular problem, is increasing, CHEMISTRY OF CHLORINATION Disinfectant capabilities of chlorine depend on its chemical form in water, which in tum is dependent on pH, temperature, organic content of water, and other water quality factors Chlorine is used in the form of free chlorine [e.g., chlorine gas] or as hypochlorites (c.g., NaOCl and Ca(OCI)3]. Chlorine applied to water cither as free chlorine or hypochlorite initially undergoes hydrolysis to form free chlorine consisting of aqucous molecular chlorine, hypochlorous acid and hypochlorite ion. Cl; + H,0 = HOCI + HY + cr ay NaOCl ocr + Na® 2) ‘Ca(OCl)2 20Cr + Ca’? Q) HOCI = H+ "OCT 4) The relative proportion of th:se “free chlorine” forms is pH and temperature-dependent. Figure | shows a distribution diagram for the various chlorine species (Cl:, HOCI, OCT) over a broad pI range, Clz can be significant only at low pll values (below pli 2); while HOCI is dominant between pH! 3 and 6, Between pil 6 and 9, the relative fraction of HOCI decreases, while the corresponding fraction of OCI increases. The dissociation of HOCI is also (eit ey se ji pH Fig 1. Distribution diagram for chlorine species at 25°C (Total Chlorine=10"M) Dr. M. Ashraf Ali, Associate Profe. or, Department of Civil Engines ing, BUETMote 1otio.ts RM," os 19 1 1o| 3] 8 e ig | 37 | 34 3 z3 2 { Compined chorine tesiduols 19 oredam.name MOI eS 8 Sais ae Chlorine osaie, >. ster Fig 2. A residual chlorine curve showing typical breakpoint (NIj-N=1.0 mg/L) b. Chlorine first reacts with reducing agents such as H2S, Fe*’, Mn’* and develops no measurable residual as shown by the portion of the curve from: A to B (sce Figure 2), ©. Addition of chlorine beyond point B results in forming mainly mono- and di-chloramines. With mole ratios of chlorine to ammonia up to 1:1 {i.c., ClNHy-N = 1:1], both mono and di-chloramines are formed. Chloramines thus formed arc eflective disinfectants and are shown as combined available chlorine residual in Figure 2 (from B to C). . Further increase in the mole ratio of chlorine to ammonia result in formation of some trichloramine and oxidation of part of ammonia to Nz and NOy (see Eq. 8. 9). These reactions are essentially complete when 1,5 mole of chlorine has been added for cach mole of ammonia nitrogen originally present in water (i.c., Cl:NHy-N = 1.5:1). This is represented by the portion of the curve from (to D (see Figure 2) Addition of chlorine beyond point D would produce free chlorine residuals and is referred to as “breakpoint chlorination”. In other words. chlorination of water to the extent that all ammonia is converted to Nz or higher oxidation state is referred to as “breakpoint chlorination”. Brcukpoint chlorination is required to obtain a free chlorine residual for better disinfection if ammonia is present in a water supply. While five chlorine residuals have good disinfecting powers, they are usually dissipated quickly in the distribution system. For this reason, final treatment with ammonia if often practiced to convert tree chlorine residuals to longer-lasting combined chlorine residuals. The difference between the amount ‘of chlorine added to the water and the amount of residual chlorine (ic. free and combined available chlorine remaining) at the end of a specified contact perio! is termed as “chlorine demandtemperature dependent. The effect of temperatuie is such that ala given pH, the traction of {OCI will be lower at higher temperatures, Generally, the disinteetant capabilities of HOC re greater than that of OCT, especially al short contact times (Muutgemery, 19851 Reactions with Impurities in Water: Reactions with Ammonia: Free chlorine reacts readily with ammonia and cert nitrogenous compounds to formscolictively known as “combined chlorine”, The inorganic chte consist of three species: monochloramine (NICH), dichloramine (NUCI:) and tachloramine oF nitrogen trichloride (NCI3), depend on a number of factors including the ratin of chlorine to ammon dose, temperature, pH and alkalinity. nines he presence and concentrations of these combined forms nitrogen, chlorine Niky | HOCI = NEC) ILO (5) NICL + HOCI =NHCh + 10 (6) NCI + HOCI =NCh + 10 7) In addition to chlorinating ammonia, chlorine also re products (c.g, nitrogen gas and nitrate) as shown below. to oxidize ammonia to chlorine-free 3Cly © 2NHy = Nig) + OH + OCT (8) 4Cl; + NH + 3H30 scl + NO. 1 OT (9) The mono- and dichloramines have significant disinfecting power and are therefore of interest in the measurement of chlorine residuals. Combined chlorine in water supplies may be formed in the treatment of raw waters containin: ammonia: chlorinated wastewater effluents. 48 well as certain chlorinated industrial effluents normally contain only combined chlorine Reactions with Other Impurities: Chlorine combines organic compounds thus increasing the chlorine cd chlorine is available to accomplish disinfection with various reducin: ts and mand which must be satisfied before Fe?", Mn", NOz’, and HS are examples of inorganic reduc nls present in water supplies that will react with chlorine. Chlorine can reaet with phenols to produce mono-, di-. or trichlorophenols, which can impart tastes and odors to waters. Chlorine also react with humic substances present in water to form trihalomethanes (THIMs. ¢ chloroform. bromoform, etc.) which are suspected human carcinogens (Note: According to USED. tpaxinwum allowable level of THMS in drinking water is 100 jeg'L) Break Point Chlorination: Ifchlorine is added to water containing reducing agents and animonia (either naturally present or added to water to produce combined chlorine), a hump-shaped breakout curve 1s produced as shown in Fig. 2. The different segment of the curve is described helow a, If the water is free of ammonia and other compounds that miiy ieact with chlorine, the application of chlorine will yield free available chlorine resulual of sume concentration This is denoted by the “no demand tine™ wt the “zero demand hin ue 2)The procedure to be followed in the laboratory is described below (a) Place 200 ml of the sample in an Farlenmeyer flask. Chill to 20°C or lower by immersing the Mask in a bath of ice water (b) Add a crystal of potassium inline and Tmt of concentrated bydrochlorie acid to the water. (c) Add 1 ml of starch solution (60) t the water. A blue color shows the presence of chlorine in the water. (d) Titrate the solution with standard sodium thiosulfate (Nay the blue color just disappears. (e) Record the quantity (in mL) of sodium thiosulfate (NasS:0.) solution used :O.) solution (68) until CALCULATION: Residual chlorine (my/L) = mL of 0.001N Sodium thiosulfate used \ Multiplying | actor (ME) where, MF = normality of NazS0) x equivalent st, af Cly x L00H "(ml of sample taken) = 0.001 x 35.45 x 1000/ 200 = 0.1773 ASSIGNMENT QUESTIONS (1) What are the major disadvantages of chlorination? Name some of the altemate Q) Q disinfcctants, You would like to perform chlorination to a water sample with pl 7.5. At this pH, what would be the relative proportions of HOCI and OCT (sce Fig. 1). What kind of change pH would you propose in order to increase the relative proportion of HOCI, whieh is a better disinfectant. Schematically draw a “chlorine residual” versus “chlorine dose’ with no ammonia or organic matter. curve for a water sample Sar te oe oneModule 4 ue oduie 4(a)Cont. aS ‘Test for Thermotolerant (faecal ) Coliforms by Membrane ation (MF) Method ‘STANDARD PHOTOSTATE KUET MAINGATE [ Courtesy: WHO Guidelines. for Drinking -Water Quatity,2" Edition. Volume-3, 1997] 1 Principle In contrast to the multiple-tube method, the membranc-filtration method gives a direct count of total coliforms and thermorolerant coliforms present in a given sample of water. The method is based on the filtration of a known volume of water through a membrane filter consisting of a cellulose compound with a uniform pore diameter of 0.45 of 0.2)1m; the bacteria are retained on the surface of the membrane filter. When the membrane containing the bacteria is incubated in a sterile container at an appropriate remperature with a selective differential culture medium, characteristic colonies of thermotolcrant coliforms develop. which can be counted directly. 2 Volume of water sample for filtration Since the fileration area is relatively small, it can support the growth of only a limited number of colonies: the optimum number is berween 20 and $0, with a maximum of 200. If this figure is exceeded, very small atypical colonies or superimposed colonies may develop, or there may be growth inhibition due co overpopulation. The choice of the volume of sample to be filtered will depend on the type of water. Examples of typical volumes are provided in Table 3 Equipment and glassware In addition co the basic equipment and glassware used in the multiple-ube method the following items are needed for the membrane: filtration technique: + Membrane-filsration apparatus. inchiding an electric or hand-powered vacuum pump, a vacuunt flask (e.g. an Erlenmeyer side-arm flask), and a filter support. + Reusable Petri dishes. made from glass or metal iepeshls plastic Petri dishes may also be used). « Aldabeniea Fareed foe BCine ip mene RItETe + Reusable (autoclavable) boteles. for culture media (¢.g. 25-ml polypropylene bottles). Aa AW1SOLOHd GUVGNVLS* A magnifying lens. with X4 or X5 magnification for examining and counting the colonies on the membrane filters. * A boiling bathlpan: if filtration apparatus is to be disinfected in boiling water between analyses. + Sterile pipette \ml and 10m. + A graduated cylinder. 100ml. In addition to the consumables needed for the MPN, the following are required: + Membrane filters. 47-50 mm in diameter, with a pore diameter of 0.45um Singly packed, presterilized membrane filters are very convenient. Unsteril- ized membrane filters can also be used, however, and should be wrapped in paper packets in convenient numbers (depending on the number of water samples to be tested). These can then be sterilized in the autoclave and dried by rapid exhaustion of the steam. + Nutrient absorbent pads. filter-paper discs about 1mm thick, with the same diameter as the membrane filters. * Culture media. different types are available + Wax pencile for labelling Petti dishes. * Polythene bags, for wrapping Petri dishes if a dry incubacor is used. to prevent drying of the sample and media. 4 Culture media and dilution water Various media can be used for the examination of coliform organisms by the membrane-filtration method. Of these, lactose Tergitol! agar, lactose TTC Tergitol agar, and membrane lauryl sulfate lactose broth may be used for coliform organ- isms at 35 or 37°C and for thermotolerant coliform organisms at 44°C or 41.5°C. Membrane faecal coliform (MFC) broth should be used only at 44 0¢ 44 thermotolerant coliform counts. Although the use of all these media for the de- tection of presumptive coliform organisms is based on the fermentation of lactose, the characteristic reaction varies with each medium, as shown in Table — | Although it is possible to prepare the media from the basic ingredients, this may be impractical in a small laboratory. The use of dehydrated media is there- fore recommended. The media can be prepared as a broth and used together with nutrient absorption pads, or as solid agar plates. The broths may be solidified by the addition of 1.2-1.5% agar before boiling. 5 Procedure The procedure generally used is described here, but differenc cypes of filtration units and equipment exist. : May-25STANDARD PHOTOSTATE KUET MAINGATE apuoIyD wryozeNsIAvaudi-C'E'Z > (OHM 10 OSI a s12npoud ssaun jo 1uaWeSopuB LE ‘2/8UNW)OD aIaeV'ERe SIONPOVD B|GeINS JO SaidWexs B7e OdsB1 PUE Z 10 Poulew voweAY auequey 2/0 VOTE/ewNUS PuE VOIDIEg "0561 I-BOLE OS! 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Connect the Erlenmeyer (side-arm) flask to the vacuum source (turned off) and place the porous support in position. If an electric pump is used, itis advisable to put a second flask between the Edlenmeyer flask and the vacuum source; this second flask acts as a water trap, and thus protects the electric pump. B. Open a sterile Petri dish and place a sterile absorbent pad in it. C. Add broth medium to sat- urate the pad; remove excess broth. wo 8525 Wad 96625, wo 96827 May27D. Assemble the fileration unit by placing a sterile mem- brane filter on the porous sup- port, using forceps sterilized by flaming. E. Place the upper container in position and secure it, (The type of clamp used will depend on the type of equipment.) F. Pour the volume of sample chosen as optimal for the type of water into the upper container. IF the test sample is less than 10 ml, at least 20ml of sterile dilution water should be added to the cop container before filtration. Apply the vacuum. 0 9628 10 366529, 0 98650 STANDARD PHOTOS KET MAINGATEG. Take the filtration unit apart and, using the sterile forceps, place the membrane filter in the Petri dish on the pad with the grid side up, Make sure that no air bubbles are trapped between the pad and the filter. g : H. Leave the Petri dish at room temperature oF at 35 or 37°C for 24 hours, for resuscitation of stressed microbes. x a I. Place the dishes in an in- cubator ar 44 + 05°C for 18-24 hours with 100% humid- ity. Alternatively, tight-fitting or sealed Petti dishes may be placed in waterproof plastic bags for incubation no 35539 2-4 hours fa)-29STANDARD PHOTOSTATE KUET MANGATE J. Submerge the bags in a wwater-bath maintained at 44 = 0.5°C for 18-24 hours. The plastic bags must be below the surface of the water 24 hours throughout the incubation period. They can be held down by means of a suitable weight, eg. a metal rack. Wo 38508 The colonies of thermotolerant coliform bacteria should be identified from their characteristics on the medium used. The number of thermotolerant coliforms per 100ml is then given by: Thermotolerant coliforms per 100ml _ no. of thermorolerant coliform colonies counted no. of ml of sample filtered sis 4(a)-30w Do Coliforms sok When Cultured? Total Coliform : This is how a total coliform culture might appear. After proper sampling and filtration, the bacterial retentive membrane filter is placed on top of ME-Endo media containing lactose, protein di- gest, vitamins elective chemicals, and Schitt's Reagent. As'the membrane incubates for 24 hours at 35°C + 0.5, the media diffuses through allowing only coliforms the pores in the filter, supplying nutrients to the multiplying bacteria. Many kinds of bacteria the water sample can grow and form col- e conditions, bit only the Coli- S| nt lactose. One by-product of this reaction is an acid aldehyde complex that will combine with the Schiff's Reagent to form an iridescent green coating over the growing col- onies. Thus, the coliforms can be identilied as wihen seen with 10-20% magnilicalion ‘Tuorescenrimuminalion. ULe64i Colitorm This is how a fecal coliform culture might appear. The liltation step is similar to that for total colt- form.“The media used for this test is M-FC, con- taining lactose, protein digest, vitamins, selec- tive chemicals, and aniline blue dye.The.mem-— brane is incubated for 24 hour $C = 02, fecal origin to growinto visible colonies. The non-fecal coliforms, due to heat shock, will not grow’ As the fecal coliforms grow, they ferment lactose, producing acid which reacts with the aniline dye to produce a blue color. When viewed with a stereomicro- ‘scope at 10-20X magnilication, all colonies STANDARD PHOTOSTATE KUET MANGATEentesic wae What is a Coliform? The coliforms are a group of gram-negaiive. tod-shaped, lactose fermenting bacteria native to the intestinal ract of mammals. They are ‘members of the family of bacteria called En- lerobacteriaceae and, like the other members, ‘can be cultured from mammalian feces. “Total coliform relers tospecies ol Escherichia, | Ka ferobacter, Serratia, Cirabacl ‘and Edws lof thee species, except scherichia, can orishas iee-lvi ies ‘aswell as Being ialeslinal organisms (rel. 1). Fecal coliform defines primarily Escherichia coli, and. si ‘Sf are “distinguished from ihe total coliform by having ‘capability of fermenting igor ateavates Face eeere os wellap al S50. te optTMUTTIOr eee ee vasa tneenuicaiec “Zo.zhas been shown to be the best tempera- {ute to select for coliforms speciticaly cl fecal origin. Any total coliform count may include fecal ccolforrh organisms. ——_— Why Test forColform? > Epidemics such oid lever, dysentery and ‘Cholera are caused by pathogenic bacteria Fenaenied vs pouied ani water Te “pela are ONCUTTScuTe vi rhaena ningiinal aaragiian sed nnieae 40 half. "Fela ustream Hiartbanl We Uk {o.delect their presence in orderto determine. alee ornate pra erate OOH Wor tmomseives, that cont me relatively hatmiess ruin : 2) are Ylalfery ay To isolate and because They normaly sue fonger than The i “Producing atoanss.c0 ilorins are @uselul ‘idicator of the possibie presence of entenc -Danogenic baclesia and viruses imost cases. ree of disease-producing back "ecal collorm analysis is a1more Welinitive for recent lecal pollution than the total color test, and facal coliform is the standard test or- {ganism used in many laboratories testing treated sewage, untreated public waer supplies, and such primary contact waters as ‘swimming areas. ‘Gat ma A832 Uren Pets USA Cena COPE RSC ECRKE amy Why the MF Method? Y STANDARD PHOT KUET NANG ‘The wide acceptance ol ihe Membrane Fillion (MF) technique 1s documented in the folowing {quotation from Standard Methods for he Exam- ination ol Water and Wastewater (rel. 3, page 928) ‘7'Since publication ol the 11th Edition, wide~ spread use of the (membrane filtration) technique has confirmed its value, especially its high degree of reproducibilily, the possibility of testing relatively larger volumes ol sample. andits | ~ ability to yield definite results more rapidly than the standard tube procedure ‘Public Health Service Drinking Water Stan- _daids (fevised 1962) recommend the mem- ‘brane iter alysis oLinterstate wwatets. The following advantages are cited by the National Training Center of the U.S. Environmental Protection Agency (EPA). sane ie oe tea aha oe pours ss youre as Coripared wi a6 86 hours ots ertaes eeeantaTenToeTreTOS 2. Much larger and ence move representative, Fampiesolwair ean be sampled routinely win mame ier. 2. Numeneat results rom membrane ters Have much greater precision (reproducibility) than is expected with the lermentation tube method. 4, The equipment and supplies required are not bulky A great many samples canbe examined with minimum requirements tor laboratory Ee oseetecistans Drinking Waler Supplies”. the EPA slates that the MF method is preferred because il permits analysis ol large Sempre worOmes wn reduced intron et 10) ST pnee aE oe Oca st asimuch ashe MPN tetnod per test, Tvs freantconsigerable savings fo any iaboralor ‘oer pailoiningcomerenwress The ulls ol bacterclogeca walt tesing canalen be invandated by celaysin process ing For this reason, Standard Methods (ref 3. page 907) recommends held testing when there + would otherwise be more than a six-hour delay between sampling and processing, The MF ‘method is the only approved method that lends itsell 10 held testing anoes CONSULTANCY RESEARCH AND TESTING SERVICES (CRTS) DEPARTMENT OF CIVIL ENGINEERING - Khwina University of Engineering & Technology, Barigladesh ST ‘ANDARD acres rey AGN WHT 20 a AMAT, ei KUET MANGATE ‘ : fae Strainer Design for Deep Tube wel Seipel 30 Sub-Soil ] - | ae ayers a) { Dee(mm) | Pom | Ue Ove? View 255-265, = 07 2.38 |For’ the selected : [265-275 0330 18 J aawiter layer (265t - BISANS [eg] to 2st: fs 285-295 [240- ] Diy 025mm: . 295-305, re [2307] Dw 42mm: | 315-325, 268 | Ue as ; 269 aig | 234 Orupicial grovel 281 | packing iskeynired | 2A5 | Nate" All the sub-soll samples were spl by he eli Water Requirement’ 4000 gal/h = 66.67 wn (as per nequirciment af the eliet» (30% of gravel pachV/30% of: 4 h size distibutiontiieve of 315-325 1 uly layyrs DO 28MAl lll, ort Cami a Retained Zone Sizetmm) | Yb pi a 476 | 1008 00 : 0 833 Sark 120 past te 583 65-70 re sige, | ry 1000 | 70 i 4 ain size distrtution curve of Gravet Pack Materials, yy 0.64 nm Slot No = (0.64/25.4)x1000 = 25.2 = 20 No. (Hom diwsnsen's Table 3 Using 4 in. dia. strainer, Open aren = 26 in" Countersigned by { — Testing OMeet t z i i ki oN ‘oara \3\CONSULTANCY RESEARCH AND TESTING $ERVICES (CRTS) aS. DEPARTMENT OF CIVIL ENGINEERIN - Khulna Univesity of Engineering & Technolegy, Bangladesh eho: XNT- 70st 201, Frege 8-7 77S. a at et ae Yield Caleutation; Yield 26 0.31» 8.06 yprv/l. Using RS. = 20, Yield ~ 8602.0 ~ 4.03 gpm Now for 20 f sffainerlengah of dey 28S (49 305 A, = ‘Tal Yield = 4,03 gpm x 20 10 Si“ pn 2664607 jpn OK, i Well Lox, : é : ‘ Bore Log Depth aay Tadeo Gravel pick Medium Sand 265 0 Medium to Coarse Sand | = i 285 H1-E- xi 285 8 Blea Dia ach Lem 20 | Maser | QRS MOSH | =I 32 | U Be i280 | Fine to Medium Sand ws f ; ssn Countersigned by Jest Performed hy i i% Finer % Finer CONSULTANCY RESEARCH AND TESTING SERVICES (CRTS) DEPARTMENT OF CIVIL, ENGINEERING + Khulna University of Engineering & Technology (RUE), Kiuutna-9208 Tel: 041-769471-5 (2Q), 201); Fax: 041-774780. Gradation Curve of Sand (BH Depth= 255-265ft) D,=0.40mm; D,,=0.17mm Pes crap ah a ee Sica ad | | | | | | | 1 Sieve Opening (mm) Gradation Curve of Sand (BH Depth= 265-275 ft) Samm; D,,=0.30mm . Le nang gl fe eam] 10 1 on oor Sievé Opening (mm) * ‘Countarsigned by < Test performed byDEPARTMENT OF CIVIL ENG Khulna University of 1 “Vel: 041-769471-5 (200 fs \ Depth, 315-325 f 5 Lew a = © é z 4 I Gravel Pach | Fa Countersigned by x = \