materials and methods - shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/7193/12/12_chapter...

CHAPTER – 3

MATERIALS AND METHODS

Materials and Methods 29

MATERIALS AND METHODS

The present study is aimed to analyse the existing pattern of water quality of river

Periyar and to assess its impact on haematological parameters and survival rate of the

selected fishes (P. sophore and H. brachysoma) in Periyar. The increased number of

industries on the banks of the river discharges the treated or partially treated or untreated

effluents into the water body. These discharges along with domestic waste and seepage

from septic tanks are responsible for the chief assault on the ecological system in the

receiving waters. When these waste materials are added to natural waters the effect on

the water quality depends entirely on the interaction of specific wastes with the delicate

balance of all the physical, chemical and biological factors of the natural aquatic

ecosystem.

3.1 Study Area

For the water quality assessment of Periyar, sampling and field studies were

carried out periodically at five locations (Collection Sites I to V) from Boothathankettu in

the upstream to Eloor (Fig. 1).

Collections of water samples were carried out during pre-monsoon, monsoon and

post-monsoon periods from April 2002 to December 2004 (April 2002 pre-monsoon, July

2002 monsoon, December 2002 post-monsoon, April 2003 pre-monsoon, July 2003

monsoon and December 2003 post-monsoon, April 2004 pre-monsoon, July 2004

monsoon and December 2004 post-monsoon). Water samples from different sites were

analyzed for physico-chemical parameters and the fishes were treated in polluted water

(Site-V) at varying concentrations for different durations to study the stress effect on the

haematological parameters. The mortality rates of the experimental fishes were recorded.

The rainfall data for the respective years were also noted (Table. 2 and 3).

The sampling sites were selected based on un-polluted area (upstream of the

river- Sites 1 to IV) and polluted area (down stream, Eloor industrial belt region - Site-V).


3.1.1 River Periyar

River Periyar is considered to be the longest river in Kerala, traversing 244 km

within the state (PWD, 1974; CESS, 1984). The river has a length of about 244 km and a

catchment area of about 5284 km2. The river originates form the Sivagiri hills of about

1830 m elevations above MSL and flows through all the three physiographic provinces -

the high lands (>75 m), midlands (8 - 75 m) and lowlands (<8 m) (main features of river

Periyar are shown in the Table -1). At Aluva the river bifurcates into Marthandavarma and

Mangalapuzha branches. The Mangalapuzha joins Chalakudy puzha and falls into the

Arabian Sea. The Marthandavarma splits into two, the major branch flows through the

industrial area (Eloor) and the minor goes through Manjummal. At Varapuzha both join the

Arabian Sea through the Cochin backwater system. Twenty five per cent of the state’s

industries are located along the banks of river Periyar in the Eloor region. These factories

depend on the river for uptake of water and disposal of effluents.

3.1.2 Collection Sites

Site I: Bhoothathankettu

Bhoothathankettu, situated east of Kothamangalam on the Aluva - Munnar route,

is perhaps the oldest irrigation project in Kerala. This area is already recognized as a

tourist centre due to pristine beauty. It belongs to Kuttampuzha panchayath, through

which two branches of Periyar flow (Plate 1. 1).

Site II: Paniely Poru

Paniely is about 8.5 km downstream of the barrage at Bhoothathankettu in

Ernakulam district. The site is accessible by motorable road, both from right and left bank

of the river. The left bank approach is from Kuruppampady near 21 km in the Aluva –

Munnar road. The site chosen is easily accessable at all times of the year except during

unusual heavy floods. The river at Paniely branches into two, forming an island in

between. The upstream side of the island is completely exposed rock. The entire island

will be submerged during unusual floods. (Plate 1. 2).


Site III: Kalady

Across the river at Kalady is the Sree Sankaracharya Bridge. Water is drawn for

the purpose of irrigation as well as drinking. The drinking water for Angamaly town is

drawn from here. Water scarcity is experienced by substantial part of the population here.

Small deltas have also been formed during recent years in the river at Kalady due to the

ruthless unauthorized sand mining. Small patches of islands have also been formed at

different areas of the river, which would become more prominent during summer (Plate 1 -

3).

Site IV: Chowara

It has a river boundary of 4km length. About one thousand households are settled

on the river banks. Very heavy sand mining is going on here, as the water level is very

low. There is one irrigation unit situated near the location besides a ferry (Plate 1. 4).

Site V: Eloor (Industrial area)

Eloor is the most industrialized zone of the river. Many hazardous industrial

effluents emerging from the nearby factories seriously threaten Eloor, situated on the side

of river Periyar. There are around 250 factories in this area, 125 of which are chemical

factories. Most of these industries are over 50 years old and are most polluting. The

industries take in large amounts of freshwater from the river and inturn discharge

concentrated effluents with nominal treatment. This leads to the large scale devastation of

aquatic life in the river and the farmlands in the region. There are more than 30 effluent

pipes spewing toxins into the river directly from the industry. Such criminal practices of the

companies in this largest industrial area of Kerala are jeopardizing the sources of drinking

water and food of the aquatic communities (Fig. 2 & 3), (Table. 4), (Plate 2 to 4).

3.2 Experimental Fishes The fishes selected for the study are Puntius sophore (Hamilton) and Horabagrus

brachysoma (Gunther). Puntius sophore comes under the family Cyprinidae of the order


Cypriniformes and Horabagrus brachysoma belongs to the family Bagridae of the order

Siluriformes. (Plate 5 A and B) (Jhingran, 1991).

Puntius sophore (Hamilton)

Colour in live state is beautiful silvery, back grey-green to brownish flanks with a

somewhat bluish lustier, underside white, a deep black round blotch at the base of caudal

fin, a similar black blotch on the central part of dorsal fin or also on the anterior part of

body adjacent to dorsal fin. Fin hyaline in mature females, anal and pelvic fins brick red in

mature males during breeding season. It is well known that they are particularly

susceptible to the oxygen content of the water and give a clear index of the state of water

(Srivastava, 2002).

Horabagrus brachysoma (Gunther)

Body is naked, not covered by scales. Mouth is sub terminal, and the jaws

containing viliform bands. Barbels two pairs, maxillary barbels extend posterior to pectoral

fin base and others shorter. Rayed dorsal fin is present with a strong, feebly serrated

spine. The adipose fin is short and the caudal fin is deeply forked. Color in live is greenish

yellow above, a deep round blotch on the shoulder surrounded by a light yellow ring.

Dorsal and anal fin are yellowish at their margins. Caudal fin is yellow with a semi lunar

thick black ring at caudal fin base in many cases (Templeton, 1995). This fish is endemic

to Kerala, which is being well relished as an important table fish.

3.3 Water Analysis

Surface water samples were collected in clean large polythene cans from the

selected five sites of the river Periyar using a country boat. The collected samples were

brought immediately to the laboratory to test the various physical and chemical

parameters. To estimate oxygen content, the samples were fixed in the BOD bottles at the

sites itself soon after collection. Standard procedures (APHA, 1998) were used for the

analysis of the physical and chemical parameters of the water samples. The mean values


of the results were calculated, which were then tabulated and graphically represented. The

results obtained were also compared with the standards (Appendix - I).

3.3.1 Physical parameters

Temperature (oC)

Temperature was measured at the sampling station itself, using centigrade thermometer.

Total Dissolved Solids (mg/l)

It was determined as the residue left after evaporation of the filtered sample.

Total Suspended Solids (mg/l)

Total Suspended Solids include insoluble matter. It was determined as the non-

filterable residue left after filtration of the sample.

3.3.2 Chemical parameters

pH

pH was measured using pH meter.

Total Alkalinity (as CaCO3 mg/l)

This is the measure of the capacity of the water to neutralize a strong acid. The

alkalinity in the waters is generally imparted by the salts of carbonates, bicarbonates,

phosphates, nitrates, borates, silicates etc., together with the hydroxyl ions in free state.

Total alkalinity was estimated by titrating the sample with a strong acid.

Acidity (as CaCO3 mg/l)

Acidity is due to the presence of uncombined carbon dioxide, salts of strong acids

and mineral acids. Acidity was estimated by titrating the sample with a strong base such

as NaOH using methyl orange or phenolphthalein as indicator.

Turbidity (NTU)

Turbidity is an expression of the optical property that causes light to be scattered

and absorbed rather than transmitted with no change in direction or flux level through the


sample. It was measured with the help of Nephelometer (Turbidity meter) and expressed

as Nephelometric Turbidity Units (NTU).

Total Hardness (as CaCO3 mg/l)

EDTA titrimetric method is used for the estimation of hardness. It is generally

caused by the calcium and magnesium ions present in water. Polyvalent ions of some

other metals like strontium, iron, aluminium, zinc and manganese etc. are also capable of

precipitating the soap and thus contributing to the hardness.

Dissolved Oxygen (DO) (mgO2 /l)

Dissolved oxygen was estimated by Winkler’s Iodometric method.

Chemical Oxygen Demand (COD) (mgO2 /l)

This determines the amount of oxygen required for chemical oxidation of organic

matter using a strong chemical oxidant such as Potassium dichromate. It represents

chemically oxidisable pollution load present in water and hence a reliable parameter for

judging the quantum of organic pollution in the aquatic system.

Chloride (mg/l)

For the determination of chloride, Argentometric method was used. Silver nitrate

reacts with chloride to form very slightly soluble white precipitate of AgCl. At the end point

when all the chlorides get precipitated, free silver ions react with chromate to form silver

chromate of reddish brown colour.

Nitrates (mg/l)

Ultraviolet spectrophotometric screening method was used for the determination of

nitrate.

Heavy metals

Aluminium (ppm)

Direct Nitrous Oxide – Acetylene flame method was adopted.


Iron (ppm)

Iron usually exists in natural water both in ferric and ferrous forms. It is converted

into ferrous state by boiling with hydrochloric acid and hydroxylamine. The reduced iron

chelates with 1, 10 - phenonthroline at pH 3.2 to 3.3 to form a complex of orange-red

colour. The intensity of this color is proportional to the concentration of iron and follows

Beer’s law, and therefore, can be determined using spectrophotometer.

Cadmium (ppm)

Anodic stripping voltammetric method was used.

Zinc (ppm)

Zinc was determined by Zincon method.

3.4 Experimental Details

The fishes were collected from river Periyar and brought to the laboratory without

mechanical injury. They were acclimated to laboratory conditions in cement tank for at

least 15 days prior to commencement of the experiment. Then the fishes were moved to

glass aquaria, where they were again acclimated for 7 days. Fishes were fed with Kijaro

floating pellet and groundnut oil cake twice a day. The cement systems and glass aquaria

were cleaned and water replenished once daily, in order to eliminate metabolites and

faecal detritus. Finally, individuals were transferred to experimental glass aquaria.

Measurements of the fishes were taken before the experiment.. The total length of the

fish was noted from the tip of the snout to the end of caudal fin and weight was taken

carefully using weighing balance. Average length, width and weight of the fishes were

showed in the Table 5 and 6.

The control fishes were treated with upstream water and the experimental fishes

were exposed to different concentrations of effluent contaminated water collected from

Site-V (Eloor industrial area) for various durations. The concentrations of effluent

contaminated water used to treat P. sophore were 5%, 10%, 15%, 20%, 25%, 30% and


H. brachysoma 10%, 20%, 30% 40%, 50%, 60%, 70%, 80%, 90%, 100%. Batches of ten

healthy fishes were used for the experiment.

Blood samples were collected from the control and experimental fishes directly

from the heart using disposable plastic syringe. The blood was kept in two vials, one

coated with anticoagulant EDTA for blood count and the other uncoated used for other

parameters. Blood samples were stored in a refrigerator and analyzed within 24hrs. Blood

samples were collected from P. sophore at the interval of 1, 2,4,6 hours of treatment and

H. brachysoma at the interval of 12, 24,48, 72 and 96 hours. The mortality / survival of fish

in the aquaria were recorded.

3.5 Analysis of Haematological parameters

Standard methods were used for the estimation of haematological parameters:

blood smear (Hesser, 1960), total erythrocyte counts (TECs) (Hendricks, 1952) and total

leucocytes counts (TLCs) (Shaw, 1930). Haemoglobin content was measured

spectrophotometrically as per the diagnostic protocol of Boerhinger Manheim GmbH using

the diagnostic kits made by BMK Laboratories. The mean values of the results for the

various parameters were found out, then tabulated and graphically represented.

Blood Cell Count

Red Blood Corpuscles (RBC) were counted by Neubauer double haemocytometer

using Hymen’s and Turk’s solution as diluting fluid described by Dacie and Lewis (1977).

White Blood Corpuscles (WBC) differential count was carried out after staining the smear

with Wright stain.

Haemoglobin

The Hb content of blood can be determined accurately by spectrophotometry. The

Haemoglobin is first converted to methaenohaemoglobin by potassium cyanide. The

optical density of the solution is read in a spectrophotometer at 540 nm or in a calorimeter

using a yellow green filter. A calibration graph is prepared when the procedure is first


started. For the preparation of calibration graph we should have a known concentration of

standard. These standard solutions are obtained commercially (Drabkin et al., 1932). The

wavelength scale and absorbance scale may be calibrated using the mercury emission

line.

Packed Cell Volume (PCV) or Haematocrit value

PCV may be defined as the volume of packed red cells in a given sample of blood

and is expressed in percentage. It was determined using Wintrobe’s method (3000rpm).

Erythrocyte Sedimentation Rate (ESR)

Westergren method was adopted. When anti-coagulated blood is allowed to stand

undisturbed for a period of time the cells tend to sink spontaneously to the bottom,

displacing plasma, which moves to the top. The rate at which the sedimentation takes

place is called ESR.

Mean Cell Haemoglobin (MCH) and Mean Cell Haemoglobin Concentration (MCHC)

These values, calculated from the results of the RBC count, Hb content and PCV

have been widely used in the classification of anaemia. MCH is deduced from the

haemoglobin and red cell count, whilst MCHC is calculated from the measured

haemoglobin and the deduced PCV (Baker and Silverton, 1976).

3.6 Mortality Rate

Observations of the state of the experimental fishes were done at fixed intervals, at

the beginning of the experiment and at 1, 2, 4 and 6 hours in P. sophore and at 12, 24, 48,

72 and 96 hours in the case of H. brachysoma. The number of fishes alive, overturned

and dead in each concentration was recorded. From the data, mortality rates were

deduced.


Morphology of RBC and WBC

Thin blood films were prepared, immediately after collection, on previously cleaned

glass slide. It was then air dried, fixed in methanol and stained using Wright stain

(Chinabut et al., 1991). Photographs of the blood cells were taken using Nikon microscope

(Model - Eclipse E 400) to study the morphology and structure of the blood cells.

Statistical Analysis

The data obtained for the various parameters investigated were statistically

analysed to determine the levels of significance, employing analysis of variance (ANOVA)

following Gomez and Gomez (1986).

39

TABLE - 1

MAIN FEATURES OF RIVER PERIYAR

Origin : Sivagiri Hills, Tamilnadu

Length : 244 km

Direction of flow : Westwards

Catchment area in Kerala : 5284 km2

Average rainfall in Catchment area : 400 cm /year –1

Rate of flow: Minimum : 9.66 m3 Sec-1

Maximum : 1364.66 m3 Sec-1

Maximum Width : 405 m

Utilization

Domestic : 260 Mm3

Irrigational : 450 Mm3

Industrial : 1844 Mm3

Number of impounded Reservoirs : 14

Number of Hydroelectric Schemes : 6

Source: CWRDM (1995)

40

Table 2 Rainfall Data (mm) of river Periyar basin from October 2002 to December 2003

2002 2003

Date Oct. Nov. Dec. Jan. Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Oct. Nov. Dec.

1 3 1.4 17.2 58.3 74 5.4 6.4

2 2.3 8.2 18.6 22.8 2.4 4.2 33 21.2

3 0.5 12.4 25.4 10.5 0.2 17 84 55.6

4 0.1 65.1 12.6 8.2 15.6 35 7.1

5 0.6 42.6 20.2 47

6 20.4 2.1 37.4 8.4 1.6 88 3.2

7 65.7 3.4 5.2 14.1 64 54 13.4

8 5.4 14.2 2.8 50 3.6

9 0.6 52.8 6.2 4.2

10 23.8 0.6 8.2 18.6 5.4

11 0.3 4.4 13.2 0.2 5.8 2.6

12 12 19.4

13 2.1 25 8.9 6.7 6 4.2 1.6 12.4 3.2

14 27.3 39.2 4.2 55 36.4 12.6 0.5

15 2.4 25 4.6 10 64.2 54.2 36

16 46.1 2.4 37 20.1 65.4 3.2 32 3.1

17 21.8 3.6 28.6 47 34.8 7.1 7.4

18 5.6 1.1 14.2 9.2 6.4 10.3 7.4

19 26.8 14.2 29 24.1 4.4 3.5

20 48.2 81 0.3 3.2 26

21 32.4 48 0.7 23.6 46

22 0.3 18.8 89 12.2 105 32

23 65.4 14 36.4 48.4 6.2

24 10 37.1 34.2 10

25 7.3 28.7 5.1 12

26 37.4 5.4 20.5 2.4

27 12.1 34 36.4 1.2

28 6.5 40.3 35 39.3 3.6

29 0.2 4.6 19.6 2.4

30 1.2 125 5.8 20.2

31 3.2 7.4 4.2 Total 214 Nil Nil 29 93.6 108 385 163 534 600 611 185 621 138 Nil

(Source: Rain Gauge Station Keerampara, Periyar River Basin)

41

Table 3 Rainfall Data (mm) of river Periyar basin for the year of 2004

Months Date Jan. Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Oct. Nov. Dec.

1 5.1 1.6 33.4 3.9 26

2 9.4 2.2 4.2 1.4 31

3 11 28. 7 72.4 15.6 39.4

4 9.2 130 50.1 140 26

5 0.6 22.4 101 74.2 39.6

6 28 60.2 4.6 2.3 69.2 18 36

7 2.6 90.5 18.8 5.4 21.5 2.6

8 18.4 11.2 17.2 10.6 1.5 12

9 22.5 47.6 11.5 10.3 5.3

10 30.2 43.7 43.7 0.5 15

11 1.6 48.4 33.2 33.2 19

12 2.1 12 39 8.2 24.5 2.3 2.3 8.2 4

13 5.2 32.8 6.4 30.6 10 3

14 2.4 1.3 28.4 6.1 29.5 8.2 0.5

15 25.4 18.1 36.2 19.7 7.9

16 6.2 10.2 55 46.4 13.5 13 1.5

17 16 32.6 24.2 40.1 17.2 2.6 40

18 24.2 18.7 7.1 4.5

19 25.4 3.2 20.2 20.2 3.8 10

20 18.2 3.9 36.4 3.9

21 10 7 5.6 1.3 16

22 2.4 46.4 6.1 6.9

23 5.2 12.2 37.2 3.9 12

24 24.6 10.4 24 1.2

25 11 26.2 4.2 26.2 3.8 13 14

26 5.4 16.7 9.1 0.5 4.5

27 13 7.2 3.5 8.5 3.7 22 20

28 1.2 1.5 49.6 30 13.7 56 26

29 24.2 30 35.4 27.1 3.4 1.3

30 47 4.7 1.2 31.2 48.3 0.5 17

31 11 4 3.9

Total 24.2 2.1 113 153 630 827 679 498 197 310 Nil

Nil

(Source: Rain Gauge Station Keerampara, Periyar River Basin)

43

Fig. 2 Sketch of effluent outlets into Periyar in Eloor - Edayar industrial region

44

Fig. 3 Eloor – Edayar Industrial Area

45

Table 4 Some Major Industries located in the Eloor - Edayar region along the shores of river Periyar

SI. No.

Name and address of Industry Raw Materials Products

1 Indian Aluminium Co. L t d, (IAC) Udyogamandal (1943)

Aluminium, Pitch, Aluminium, Fluoride, Cryolic

Aluminium rode, Al, ingots Al, Extension Waste Water 4110 x 10 lit /day

2

Travancore Chemicals Manufacturing Co. Ltd (TCMC) Kalamassery (1943)

Copper Scrap, Hel, Sulphuric Acid, Bauxite, Washed Granalite, Caustic soda, Sodium Chloride

Copper Oxychloride, Copper Sulphate, Sodium Aluminate, Aluminium Sulphate, Sodium Chloride, Potassiuum Chloride Waste Water 239.08 lit/day

3

Fertilizers and Chemicals Travancore Ltd (FACT) Udyogamandal, (1947)

Sulphur, Rock phosphate, Naphtha, Hydrochloric acid

Ammonia, Ammonium sulphate, Ammonium phosphate, Ammonium Chloride, Sulphuric acid, Super - phosphate, Sulphur dioxide Water discharge 20658 x 106 lit / day

4 Travancore Cochin Chemicals Ltd, (TCC) (1951)

Sulphur, Soda, Ash, Barium carbonate, Caustic lime, Common salt.

Caustic soda, Sodium sulphide, Sodium hydrosulphate, Liquid chloride, Hel, Waste Water 3504 x 106 lit/day

5 Indian Rare Earths Ltd. (IRE) (1951)

Monozoic sand, Caustic Soda, Hydrochloric acid Chloride, Nitric acid

Trisodium earth, phosphon rare earths oxide, cerium oxide, Rare earths chloride.

6 Hindustan Insecticides Ltd (HIL) Udyogamandal (1951)

Benzene, Alcohol Chlorine

DDT, BHC, Endosulfan, Oleum.

7 Cominco Binani Zinc Ltd (CBZL) Binanipuram (1967)

Zinc, Zinc concentrate. Sulphuric acid, Cadmium

8 Periyar Chemicals Ltd Binanipuram (1965)

Sodium formate, Sulphuric Acid

Formic acid, Sodium sulphate, Waste water 844.96 lit / day

9 United Catalysts India Ltd, (UCI) Binanipuram (1970)

Alumina, Copper, Zinc, Iron scrap, Croplite, Sulphuric Acid, Sodium, Ammonia, Chloride, Carbon dioxide

Catalysts for Fertilizers and Petrochemical Industries Waste Water 125 x 106 lit/day

10 Sree Sakthi Paper Mills Ltd, Edaval

Waste paper, Case boards etc.

Craft paper, Waste Water 2200 m3 / day

11 Cropton Greaves, Athani

Tungsten, Epoxyresin, Aluminium cap.

Bulbs, Waste Water 220 m3 / day

12 Prime Agro products, I D Area, Binanipuram

Rice residue, Cereals, Salt.

Cattle feed Poultry feed

46

SI. No.

Name and address of Industry Raw Materials Products

13 Pigments India, Ltd Edayal Scrap materials Red Oxide

14 Kerala Acids and Chemicals, Edayar

Acid and other Chemicals.

Reagent preparation Waste Water 21230 m3 / day

15 Edayal Rubbers, Edayal Crumb Rubber

Rubber products for other rubber industries as their raw materials

16 Bethel Chemical Industries(P) Ltd , Edayal

Sodium silicate, Sulphuric Acid Waste Water 30 m3 / day

17 Star Refineries, Edayal, Aluva. - Waste Water 15 m3 / day

18 Algar Organic Chemicals, Manjumel Udyogamandal

- Waste Water 15 m3 / day

19 Kanyakumari polymers Edayal, Aluva. - Waste Water 62 m3 / day

20 Alwaye Techno Engg. Pvt. Ltd Edayal


21 Cochin Minerals and Rutiles, Edayal.

- Welding rods

Waste Water 50 m3 / day

22 Kairaly Tanneries (P) Ltd,ID Area, Edayal. Raw skin of animals Processed leather

23 STD Potteries, Aluva Clay, Gypsum Crockeries, Sanitary item

Waste Water 53 m3 / day

24 Auric Brown Crump, Muttom, Aluva Crumb Rubber Processed Rubber products

25 Manswill Chemicals Pvt. Ltd. ID Area, Edayal.


Source: KSPCB, 1985, 1985 A

47

Table 5 Length, Width and Weight of P. sophore used for the Experiment.

2002 2002 2002-03 2003 2003 2003-04 Body (Size)

Pre-monsoon Monsoon Post-monsoon Pre-monsoon Monsoon Post-monsoon

Length (cm) 7.2 - 8.5 6.8 - 8.3 7.5 - 8.8 6.5 - 8.2 6.9 - 8.4 7.1 - 8.6

Width (cm) 2.5 - 3.8 2.2 - 3.3 2.8 - 4 2.1 - 3.5 2.4 - 3.7 2.5 - 3.9

Weight (g) 11.3 - 18.8 9.8 - 17.8 12.8 - 20.1 10.5 - 17.6 10.3 - 16.8 12.4 - 18.5

Table 6 Length, Width and Weight of H. brachysoma used for the Experiment.

2002 2002 2002-03 2003 2003 2003-04 Body (Size)

Pre-monsoon Monsoon Post-monsoon Pre-monsoon Monsoon Post-monsoon

Length (cm) 15.2 - 22 18.8 - 24 14.2 - 23.2 12.6 - 22.4 16.5 - 24.3 13.7 - 23.2

Width (cm) 3.2 - 5.4 4.6 - 6.8 3.7 - 5.8 2.8 - 4.9 3.4 - 5.4 3.3 - 5.2

Weight (g) 35.4 - 80.2 40. - 90.5 32.8 - 84.6 28.7 - 76.5 38.3 - 93.2 33.3 - 86.4

PLATE 1: COLLECTION SITES

I. Bhoothathankettu

II. Paniely Poru III. Kalady IV. Chowara V.

PLATE 2: SITE-V INDUSTRIAL AREA

A. View of river Periyar from Eloor B. Close view of Industries near Periyar C. Industrial area of Eloor near the banks of river Periyar - I D. Industrial area of Eloor near the banks of river Periyar - II E.

PLATE 3: SITE-V INDUSTRIAL AREA (Continued)

E. Point source of effluent - 1

F. Point source of effluent - 2

G. Point source of effluent - 3

H. Colour change of water due to the effluent

PLATE 3

. -

k I.

3' I

( I .

r

. -

PLATE 4: SITE-V INDUSTRIAL AREA (Continued)

I. Spreading of the coloured effluent

J. Colour change of water due to the effluent

K. View of river Periyar 2 km down stream from Eloor

PLATE 4

PLATE 5: EXPERIMENTAL FISHES

A Puntius sophore (Hamilton) B Horabagrus brachysoma (Gunther)

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