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8/17/2019 Jurnal Pengukuran oksigen

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8 International Journal of Water Research 2013; 1(1): 7-11

made to solve the water scarcity on one end and creatingwealth from waste on other end. The main objectives ofthis study were

To investigate survival of the fish Catla catla exposed pure water, raw sugar factory effluent and recycledsugar factory effluent.To study the respiratory physiology of the fish exposedto raw and recycled sugar factory effluent water and tocompare this result with the control.To expose the recycled sugar factory effluent as a goodmedium for fish growth.

2. Materials and Methods:2.1. Collection and Maintenance:Healthy fingerlings of Catla catla collected from a localfish farm near Madurai and transported to the laboratory inclosed polythene bags filled with oxygen. Duringtransportation, care taken to reduce any hyperactivity and

physical injuries to fish. Immediately after reaching at the

laboratory, disinfective dip treatment with 0.1% KMnO 4 tothe fish as a precaution. Fish acclimatized to lab conditionof aquatic vegetation, for a period of one month. Only non-chlorinated ground water used. During this period, the fishfed with pellet diet having 38% crude protein preparedaccording to standard method (Hardy, 1980).2.2. Fish FeedThe feed for the test animals prepared with the constituent

peanut oilcake, rice bran, fish meal, dry fish and tapiocaflour in the ration 3:2:2:2:1. The biochemical compositionof the feed tested using standard method (APHA, 1975)and presented in Table 1. The energy value of theformulated feed estimated using an Oxygen bomb

calorimeter.2.3. COLLECTION AND CHARACTERIZATION OFEFFLUENT: Raw and recycled effluent from the sugar factory effluentcollected from the main points of discharge atAlanganallur, Madurai. The collected effluents transportedimmediately to the laboratory and the physical andchemical characteristics of raw and recycled sugar factoryeffluent estimated (Table 2) using Standard Methods(APHA et al., 1992)2.4. Dilution procedureThe undiluted sugar factory industry effluent collectedfrom the discharge point of the industry considered as

100% solution. From this, the selected effluentconcentrations for the experiments obtained by diluting itwith clean non- chlorinated ground water. The physical andchemical characteristics of the ground water used fordilution also analyzed.2.5. Determination of Sub lethal concentrations ofSugar Factory Effluent:In acute bioassay studies, the determination of LC50 is of

prime importance as only then sub lethal concentration forthe experimental treatment of the test organisms could bechosen. Bioassays conducted following the standard

procedure (Sprague, 1971). Survival studies on the fish of5.0±0.5g carried out as described below. The test

individuals starved one day prior to the survival test.Groups of 10 well-acclimated fish introduced intocylindrical plastic aquaria containing 10 liters of test

medium at 28±1 0c. Fish mortality taken into account atevery 24hrs for total periods of 96 h. Dead fish removedimmediately from the medium. The test medium reneweddaily with a freshly prepared one to give a constant effectof sugar factory effluent on fish. During renewal, 80% ofthe test effluent medium siphoned out and the same volumereplaced with fresh effluent medium with the leastdisturbance to fish. During the period of survival test, allfish starved and kept in a non-aerated medium.Similarly, the LC50 values were found out for the fishexposed to the recycled effluent separately using ProbitAnalysis (Finney, 1978).2. 6. Respiration: In the present study, the effect of sub lethal concentrationsof the chosen media viz., raw and recycled sugar factoryeffluent on the oxygen consumption of tilapia (3.5+0.6gm)was studied.Fish exposed to the selected sub lethal concentrations ofraw and recycled sugar mill effluent. Separate controlmaintained for each medium for comparison. At eachconcentration, three sets of fish (10 fish in each set)exposed in glass aquaria (37x30x14 cm), containing 20liters of the medium for 6 weeks. During the exposure

period, fish fed ad-libitum with prepared feed pellets once aday for 2 hours (08.00 to 10.00 hours).The food remained collected, using a siphon, immediatelyafter feeding. In all the experiments, the medium waschanged every day. The oxygen consumption rate of thefish exposed to the selected concentrations and controlestimated following the modified Winkler’s iodometricmethod with the incorporation of sodium azide withalkaline iodide in order to eliminate any interference ofoxidizing and reducing materials that may be present inthe sample.A Simple flow through system described by Beamish etal ., (1975) adopted to estimate oxygen consumption inthe present study. Non-chlorinated ground water filled ina reservoir tank after filtering through a sintered glasscolumn with glass wool. An aspirator bottle having a thinPVC tube served as a dosing apparatus (Mariotte bottle).Stock solution of the sample prepared and placed in aMarriott bottle and allowed to flow into the reservoirtank. The required strength of the sample maintained inthe reservoir chamber by adjusting the flow of the

effluent and water. A small magnetic stirrer kept inside toensure thorough mixing of the medium within thereservoir and also to keep the dissolved oxygen content ofthe water at saturation level.From the reservoir, the test solution flowed to theconstant surface chamber, the flow being regulated by astopcock. The constant surface chamber was necessary toensure a constant surface level and to regulate the rate ofoutflow throughout the experiment. Wide mouth glass

jars with stoppers served as experimental and controlchambers. Black paper with a small observation windowwas pasted on the outer surface of the experimentalchamber to avoid disturbance to the fish. The water from

the constant surface chamber was then allowed to passsimultaneously into experimental and control jars. Thismethod adopted for testing dissolved oxygen in the


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different concentrations of raw and recycled effluent andin the control separately. Control medium kept separatelyfor each toxic medium.The rates of oxygen consumption and gill ventilationestimated for the fishes exposed to the differentconcentrations in the three toxic media after every weekup to the sixth week of exposure. The fishes starved forfour hours prior to the estimation of oxygen consumptionand gill ventilation rate, in order to avoid the effects ofany specific dynamic action.During the estimation of oxygen consumption, individualfish introduced into a respiratory chamber of exactly twoliters capacity. The lid of the chamber tightly closeddown in such a way that there was no air space betweenthe lid and upper surface of the medium in the chamber.This arrangement to avoid dissolution of gaseousatmospheric oxygen into the water in the course of theexperiment. The fish allowed remaining and respiring inthe medium contained in the respiratory chamber for twohours. At the end of the experimental period the dissolvedoxygen content of the medium in the respiratory chamberdetermined by following Winkler’s iodometric method,modified azide incorporation with alkaline iodide (Welch,1948).The difference between the oxygen content of the mediumwithout fish and that with the fish gave the amount ofoxygen respired by the fish for two hours. The oxygenconsumption rate of the fish calculated by dividing theamount of oxygen consumed by the live weight of the fish.This converted to oxygen consumption rate as that perhour. The same procedure repeated for finding out therespiratory rates of fish exposed to raw and recycledeffluent. ANOVA tests were conducted using SPSS 10software package.3.Result3.1. Survival of Catla catla The physicochemical characteristics of the test mediumestimated and the findings presented in it. The survival ofCatla catla exposed to different concentrations of thechosen test media viz., control, raw sugar factory effluentand recycled sugar factory effluent studied.

Fig.1. Oxygen consumption of fish exposed to differenttypes of effluent

The mortality value of fish exposed to variousconcentrations of the sugar factory effluent, for differenttime period viz., 24, 48, 72, 96h, observed as LC50 valuesgiven in (Fig.1).The LC50 value of sugar factory effluent, which killed50% of the test individuals in 24h exposure, was 10% andit decreased to 3% when the exposure duration increasedto 96h (Table 4). This indicates that as the duration ofexposure in the test medium increased, the median lethalconcentrations decreased. One way ANOVA test indicatedsignificant difference in LC50 values in fish exposed todifferent times of exposure ( F=13.92; P


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Table 1: Proximate Composition (Mean ± SD) of the feed Ingredient Moisture (%) Protein (%) Fat (%) Carbohydrate (%) Fiber Ash Calories

Peanut Oilcake 10.77 30.42 9.80 25.40 - 14.16 9.8Rice Bran 10.15 09.62 6.20 38.60 7.15 12.20 7.5Fish Meal 2.00 26.40 18.30 32.30 8.50 13.10 16.8

Dry Fish 1.80 30.12 05.10 - - - 2.2Topioca flour 00.60 01.80 01.60 45.80 - 0.75 3.4Total = 32.1 KJ.g-1

Table 2: Physico- Chemical characteristics of raw and recycled sugar factory effluent.Parameter ISI limit Raw Effluent Recycled Effluent

Colour Colourless Yellowish brown ColourlessSuspended solids 100 1352 400Dissolved solids 2100 3990 269

pH 5.5-9.0 5.0 6.5BOD 30 1670 980

Dissolved oxygen 6.0 Nil 4.5Dissolved chlorides 600 309 200Dissolved sulphates 2200 2759 1800Dissolved calcium 7.5 835 300Dissolved nitrates 300 60 20Dissolved nitrites 300 25 0.45

Alkalinity 482 482 125The value expressed in (ppm) except pH

1977).The 96hr LC50 value for Catla catla in differentindustrial effluents have been reported; 14% dye factoryeffluent (Navaraj, 1988), 18% textile mill effluent (Haniffaand Jassentha, 1988) and 16% paper mill effluent for(Nanda et al., 2000) have also been reported. This study

clearly indicates that Catla catla is highly sensitive to sugarfactory effluent. As there is no mortality in the differentrange of concentration of recycled sugar factory effluent,the recycled effluent is taken as such for comparison in thestudy.4.2. Oxygen ConsumptionThe measurement of oxygen consumption not onlyindicates metabolic rate but also provides and index tostress condition. Change in oxygen consumption rate has

been identified as an indicator of sub lethal stress inorganisms exposed to toxic substance (Hughes et al., 1987). The initial rise in oxygen uptake may be due to theacceleration of oxidative metabolism to meet the extra

energy demand due to pollutant stress. It may lead to theacclimation of fish to the toxic medium (Haniffa et al., 1966).After two weeks, the rate of oxygen uptake decreased as afunction of increasing concentration of toxicants and alsoincreasing exposure period. This may be due to theresponse of the fish for acclimatization in the stressmedium. The reduction in oxygen consumption may be dueto the malfunctioning of gills, which have direct contactwith pollutants and effluents dissolved in the medium.David (1956) has shown that high concentrations of toxicchemicals in the water medium will coagulate the gillmucus in fish and therefore alter the respiratory

metabolism.Thus accumulation of mucus on the gills, asphyxiation andinhibition of enzyme system at mitochondrial level could

result in the decrease of oxygen consumption (Lock, 1979).Hingorani et al., (1979) supported this by stating that toxicchemicals present in the pollutants interfered withrespiration by coagulation of gill mucus and inhibition ofenzyme systems at mitochondria level resulting in the

reduction of oxygen consumption. Moreover, this findingsupports the present study on the enzyme levels in Catlacatla exposed to Sugar factory effluent.Hoar (1984) also indicated that pollutants cause alterationsin the structure of mitochondrial membrane, which preventthe intake of oxygen into mitochondria and hence, in therate of oxygen consumption. The decrease in red bloodcorpuscles and the consequent alterations in hemoglobin-oxygen binding capacity have also been suggested asreasons for drop in oxygen uptake in fish exposed to theindustrial effluents (Murthy et al., 1986). The presence oftoxicants in the medium could bind with globin fraction ofhaemoglobin of the fish and alter the physiological activity

of the body (Dhanapal et al., 1990). Thus the decreaseoxygen consumption may be due to the damage caused tored blood cells as reported by Ganapati and Alikunhi(1950).In the present study, toxicants of the sugar factory effluentmay develop mucus in the respiratory lamella and block thegills surface resulting in poor intake of oxygen as reported

by Kandeepan and Navaraj (1993). This may inhibit theactivity of 5- aminolevulinate synthetase, one of theenzymes involved in haeme synthesis (Pamila et al., 1991).Thus the reduced oxygen consumption could be a sequel togill damage or due to hypochronic microcytic anemia undereffluent stress.

The drop in oxygen consumption of Catla catla exposed toraw effluent indicates the onset of hypoxia in fish understress, which triggers metabolic pathways of fish. However,


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there is no such decrease in the oxygen consumption of fishexposed to recycled sugar factory effluent.

Table 3: Physico – Chemical Characteristics of groundwater used for dilution

Parameter Value

Temperature 28.0 CPH 7.4

Dissolved oxygen 6.1Organic nitrogen 0.3

Total hardness 2.4Calcium 3.1Chloride 4.2

Magnesium 2.4Sodium 3.8

Except pH all value are expressed in g.1 -1

Table 4: LC50 value of Catla catla at different duration inSugar factory effluent

Hours Lc 50 values (%)24 1048 0872 0696 03

Table5. Effect of different test media on Oxygenconsumption rate (mg.kg -1.d -1) in Catla catla .(Mean ± SD)

Week Control Raw effluent Recycled effluentI 605±0.57 720±0.58 598 ±0.57II 603±0.57 710±1.09 590±1.08III 604±0.57 445±0.57 589±0.57IV 602±1.03 400±1.06 596±0.57

Table 6. ANOVA for the Effect of different media onoxygen consumption rate in Catla catla

Test media on oxygen consumption rate F (1,7) ValueRaw effluent 0.16**

Recycled effluent 19.6*Table value of F (1,7) = 5.98 ; p>0.05* p

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