Aqua Manual

1
AQUACULTURE
LABORATORY EXPERIMENTAL PROTOCALS
DR.C.V.NARASIMHA MURTHY
M.Sc.,Ph.D.,M.Ed.,
P.G.DIP IN HIGHER EDUCATION,
P.G.DIPLOMA IN JOURNALISM
P.G.DIPLOMA IN PUBLICRELATIONS AND ADVERTISING
PROFICIENCY IN GENETIC ENGENEERING
ASSOCIATE PROFESSOR (CONTRACT)
DEPARTMENT OF ZOOLOGY
&
DR.V.SAILAJA
M.Sc.,Ph.D.,M.Ed.,
ASSSTANT PROFESSOR
DEPARTMENT OF ZOOLOGYV.S.U.P.G.CENTER
KAVALI
2016
Dr.C.V.Narasimha murthy & Dr.V.Salaja .aquaculture practice manual 2016-2017 ; M.Sc. Zoology Final year 3rd semester
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AQUACULTURE LABORATORY EXPERIMENTAL PROTOCALS
INDEX
S.NO. NAME OF THE EXPERIMENT PAGE

NO.
1 DETERMINATION OF pH OF WATER SAMPLES 3
2 ESTIMATION OF SALT CONTENT BY USING SALINITY REFRACTO- 7
METER
3 DETERMINATION OF FREE CARBONDIAXIDE CONTENT IN WATER 9
4 DETERMINATION OF TUBIDITY OF WATER BY SECCHI'S DISC 11
METHOD
5 DETERMINATION OF CARBONATES AND BICARBONATES IN WATER 13
6 DETERMINATION OF ALKALINITY OF WATER SAMPLES 16
7 EXPERIMENT ON DETERMINATION OF CALCIUM HARDNESS 20
8 EXPERIMENT ON DETERMINATION OF RESIDUAL CHLORINE 22
9 DETERMINATION OF AMMONICAL NITROGEN IN WATER 25
10 POND CONSTRUCTION 27
11 SOIL QUALITY ANALYSIS OF POND 36
12 WATER QUALITY ANALYSIS OF POND 39
13 ANALYSIS OF PLANKTON 44
14 BIOMETRY OF FISH 46
15 BIOMETRY OF PRAWN 49
16 STUDY OF GENERAL CHARACTERS AND IDENTIFICATION OF 55
IMPORTANT SHRIMP AND PRAWN.
17 STUDY OF GENERAL CHARACTERS AND IDENTIFICATION OF 60
IMPORTANT FISH
18 INDUCED BREEDING IN FISH 91
19 INDUCED BREEDING IN PRAWN /SHRIMP 94
20 FIELD VISIT TO PRAWN/SHRIMP HATCHERY 97
21 EXPERIMENT ON DETERMINATION OF CHEMICAL OXYGEN 104
DEMAND
22 EXPERIMENT ON DETERMINATION OF BIOCHEMICAL OXYGEN 111
DEMAND
23 DETERMINATION OF POROCITY OF SOIL 125
Dr.C.V.Narasimha murthy .aquaculture practice manual 2016-2017 ; M.Sc. Zoology Final year 3rd semester
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Experiment No.1
Date:
DETERMINATION OF pH OF WATER SAMPLES
AIM: To determine the pH of the given water sample.
INTRODUCTION
The term pH refers to the measure of hydrogen ion concentration in a solution
and defined as the negative log of H+ ions concentration in water and wastewater. The
values of pH to a little less than 7 are termed as acidic and the values of pH. A little above
7 to 14 are termed as basic. When the concentration of H+ and OH- ions are equal then it
is termed as neutral pH.
ENVIRONMENTAL SIGNIFICANCE;
Determination of pH is one of the important objectives in biological treatment of

the wastewater. In anaerobic treatment, if the pH goes below 5 due to excess accumulation
of acids, the process is severely affected. Shifting of pH beyond 5 to 10 upsets the aerobic
treatment of the wastewater. In these circumstances, the pH is generally adjusted by
addition of suitable acid or alkali to optimize the treatment of the wastewater. pH value or
range is of immense importance for any chemical reaction. A chemical shall be highly
effective at a particular pH . Chemical coagulation, disinfection, water softening and
corrosion control are governed by pH adjustment. Dewatering of sludge, oxidation of
cyanides and reduction of hexavalent chromium into trivalent chromium also need a
favorable pH range. It is used in the calculation of carbonate, bicarbonate, C02 corrosion,
stability index and acid base equilibrium. Lower value of pH below 4 will produce sour
taste and higher value above 8.5 a bitter taste. Higher values of pH hasten the scale
formation in water heating apparatus and also reduce the germicidal potential of chlorine.
High PH induces the formation of trihalomethanes, which are causing cancer in human
beings.
PRINCIPLE
The pH electrode used in the pH measurement is a combined glass electrode. It
consists of sensing half cell and reference half cell, together form an electrode system.
The sensing half cell is a thin pH sensitive semi permeable membrane, separating two
solutions, viz., the outer solution, the sample to be analyzed and the internal solution
enclosed inside the glass membrane and has a known pH value. An electrical potential is
developed inside and another electrical potential is developed outside, the difference in
the potential is measured and is given as the pH of the sample.
APPARATUS REQUIRED
1. pH meter 5. Beaker
2. Standard flasks 6. Wash Bottle
3. Magnetic Stirrer 7. Tissue Paper
4. Funnel 8. Forceps
Dr.C.V.Narasimha murthy .aquaculture practice manual 2016-2017; M.Sc. Zoology Final
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CHEMICALS REQUIRED
1. Buffers Solutions of pH 4.01, 7.0 and 9.2
2. Potassium Chloride
3. Distilled Water
PROCEDURE CHART
Switch on the ph meter

(Atleast 30 min before the test) Prepare t i e buffer
solution 4.0, 7.0 and 9.2
4.0 7.G 9.2
Calibrate the pH meter Calibrate the pH meter

to 7.0 using the buffer to 9.2 using the buffer
anb by adjusting the and by adjusting the
calibration knob calibration knob
Calibrate the pH meter

to 4.0 using the buffer
and by adjusting the
calibration knob
SAMPLE HANDLING AND PRESERVATION
Dr.C.V.Narasimha murthy .aquaculture practice manual 2016-2017; M.Sc. Zoology Final year 3rd
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Preservation of sample is not practical. Because biological activity will continue after a sample has
been taken, changes may occur during handling and storage. The characteristics of the water sample
may change. To reduce the change in samples taken for the determination of pH, keep samples at 40
C. Do not allow the samples to freeze. Analysis should begin as soon as possible.
PRECAUTIONS
The following precautions should be observed while performing the experiment:
1. Temperature affects the measurement of pH at two points. The first is caused by the change in
electrode output at different temperatures. This interference can be controlled by the instruments
having temperature compensation or by calibrating the electrode-instrument system at the
temperature of the samples. The second is the change of pH inherent in the sample at different
temperatures. This type of error is sample dependent and cannot be controlled; hence both the pH
and temperature at the time of analysis should be noted.
2. In general, the glass electrode is not subject to solution interferences like color, high salinity,
colloidal matter, oxidants, turbidity or reductents.
3. Oil and grease, if present in the electrode layer, should be removed by gentle wiping or detergent
washing, followed by rinsing with distilled water, because it could impair the electrode response.
4. Before using, allow the electrode to stand in dilute hydrochloric acid solution for at least 2 hours.
5. Electrodes used in the pH meter are highly fragile, hence handle it carefully.
PROCEDURE
Three major steps are involved in the experiment. They are
1. Preparation of reagents
2. Calibrating the Instrument
3. Testing of Sample
PREPARATION OF REAGENTS
1. Buffer Solution of pH 4.0
1. Took 100 ml standard measuring flask and placed a funnel over it.
2. using the forceps carefully transferred one buffer tablet of pH 4.0 to the funnel.
3. Added little amount of distilled water, crushed the tablet and dissolved it.
4. Made up the volume to 100 ml using distilled water.
2. Buffer Solution of pH 7.0
a. Took 100 ml standard measuring flask and place a funnel over it.
b. Using the forceps carefully transferred one buffer tablet of pH 7.0 to the funnel.
c. Added little amount of distilled water, crushed the tablet and dissolved it.
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d. Made up the volume to 100 ml using distilled water. 3.
Buffer Solution of pH 9.2
1. Took 100 ml standard measuring flask and placed a funnel over it.
2. Using the forceps carefully transferred one Buffer tablet of pH 9.2 to the funnel.
3. Added little amount of distilled water, crushed the tablet and dissolved it. Made
4. up the volume to 100 ml using distilled water.
CALIBRATING THE INSTRUMENT

Placed the electrode in the beaker containing the 9.2 stirred buffer and checked for the
reading in the pH meter. If the instrument is not showing pH value of 9.2, by using the calibration
knob adjusted the reading to 9.2. Taken away the electrode from the buffer, washed it with
distilled water and then wiped gently with soft tissue.
Now the electrode was placed in p 7.0 buffer solution and checked for the reading in the
pH meter. If the instrument is not showing pH value of 7.0, using the calibration knob adjusted the
reading to 7.0. later electrode is washed with distilled water and then wiped gently with soft tissue.
TESTING OF SAMPLE
1. In a clean dry 100 ml beaker water sample was taken..

2. Now electrode is placed in the beaker containing the water sample and checked for the
reading in the pH meter. Waited until get a stable reading.
4. The p of the given water sample was 8.84
5. Took the electrode from the water sample, washed it with distilled water and then wiped
gently with soft tissue.
TABLE
S.No Name of the sample pH value

1 A
2 B
3 C
Result
The pH value of sample A is =

Discussion :
From the above experiment we can conclude that sample A is xxxxx(acidic/basic/ neutral) and
sample B XXXXX. In the same way sample C is XXXX.
Water quality in fish ponds is affected by the interactions of several chemical components. Carbon
dioxide, pH, alkalinity and hardness are interrelated and can have profound effects on pond
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productivity. The measure which indicates whether water is acidic or basic is known as pH. More
precisely, pH indicates the hydrogen ion concentration in water and is defined as the negative
logarithm of the molar hydroen ion concentration (-log [H+]). Water is considered acidic when pH is
below 7 and basic when p is above 7. Most pH values encountered fall between 0 and 14. The
recommended pH range for aquaculture is 6.5 to 9.0.Fish and other vertebrates have an average
blood pH of 7.4. Fish blood comes into close contact with water (1- or 2-cell separation) as it passes
through the blood vessels of the gills and skin. A desirable range for pond water pH would be close to
that of fish blood (i.e., 7.0 to 8.0). Fish may become stressed and die if the pH drops below 5 (e.g.,
acidic runoff) or rises above 10 (e.g., low alkalinity combined with intense photosynthesis by dense
algal blooms phytoplankton or filamentous algae). Pond pH varies throughout the day due to
respiration and photosynthesis. After sunset, dissolved oxygen (DO) concentrations decline as
photosynthesis stops and all plants and animals in the pond consume oxygen (respiration).
Experiment No. 2
Date:
ESTIMATION OF SALT CONTENT BY USING SALINITY REFRACTOMETER
Aim: To estimate the salt content of water in field conditions by using salinity Refractometer.
Requirements:
1. Salinity Refractometer
2. Distilled water
3. Beakers
4. Glass rods
5. Filter papers
6. Water samples.
Principle:
The refractive index of water increases with increases of salt in water. By determining the
refractive index of water we can determine the salinity content of water. For determining the
refractive index of saline water salinity Refractometer will be used. By determining the critical
angle we can determine the refractive index. A drop of water i.e. The sample which we are going
to determine the salinity will be placed on the prism of the salinity Refractometer . When light
passes through this prism some amount of light is reflected and some amount of light is
refracted. If the angle of incidence is greater than critical angle then the light do not pass through
prism and it can be seen as darkness If the angle of incidence is lesser than critical angle then the
light does not pass through prism and it can be seen as brightness. The scale on the salinity
Refractometer is designed in such a way that the region of light and dark can be observed very
clearly. Basing on this the salinity is determined.
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illuminator flap with calibration bimetallic

matted surface screw strip
light source
sample measuring shadow boundary

prism and graduation
seen through
the eyepiece
Procedure adopted.
The prism of the salinity Refractometer is cleaned with distilled water and washed with filter
paper. First by placing distilled water on the prism ,the scale of the salinity
Refractomt3er is adjusted to Zero. For determining the salinity of sample a drop of sample is
placed on the prism and shutter is closed . The reading on the scale is recoded where light and
blue Colour is merged. The scale reading is noted in the table.
Precautions:
1. Water droplet should be completely dispersed on the prism

2. No air bubbles should be present in the sample
3. care should be taken in such a way that there should not be dirt or scratches on the prism.
TABLE:
S.No Name of the Salinity (ppt)

sample
1
2
3
4
RESULTS:
1. Salinity of the A sample ppt ppt

2. salinity of B sample
ppt. ppt.
3. salinity of sample collected from
4. salinity of sample collected from
DISCUSSION:
The salinity Refractometer is very useful to determine salinity in field conditions.

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In this experiment it was found that the salinity of A sample is ppt , B sample is ppt. Salinity of
water of Anapagunta is around 0 ppt. Whereas Tambalagunta water is 7 ppt. Salinity has a major
role in growth and development of fish and prawn. For fresh water carps 0- 0.5 ppt , for tiger prawn
15-30 ppt and marine fishes 30-40 ppt are required.
Experiment no. 3
Date:
DETERMINATION OF FREE CARBONDIAXIDE CONTENT IN WATER
Aim: To determination of free carbon dioxide in waste water
INTRODUCTION;
Carbon dioxide is highly soluble in water. The source of free carbon dioxide in water is
seldom that from the air phase because CO2 is a product of aerobic and anaerobic decomposition of
organic matter and it is intimately bound in the complex carbonate equilibrium.
The dissolved carbon dioxide reacts with water to form carbonic acid. C02 + H2O {;} H2CO3
which in turn disassociates to H2CO3 §;} H++ HCO3- such that [H+][HCO3-]/[H2CO3] = 3.5 x
10'7 at 18°, thus at pH 8, the ratio of carbonic acid to bicarbonate ions is only 0.286, but at pH 7 it is
0.86 and at pH 6 it is 2.86. Below pH 4.3 almost all the bicarbonate is converted to carbonic acid.
Carbonic acid can be toxic even at pH values that are not in themselves harmful. Hence, pH is not a
reliable index of dangerous C02 pollution.
Free carbon dioxide in domestic water is of no significance because it appears to have no

direct physiological effect. In fact, soft drinks and beer are highly charged with C02.
The concentrations of dissolved C02 and carbonic acid in water have a marked effect on
fish. In their migration, fish tend to respond to slight gradients of carbon dioxide tension and to
avoid concentrations of 1 to 6 mg/L.
It is doubtful if any fresh water fish can continue to live throughout the year in water with an
average C02 content as high as 12 mg/L. Concentrations of 20 mg/L will quickly prove fatal to the
more sensitive species. For example, the lethal limit for trout has been reported as 45 mg/L.
Free carbon dioxide in excess of 20 mg/L may be harmful to fish in normal fresh water, but
when the dissolved oxygen content drops to 3 to 5 mg/L, lower C02 concentrations may be
detrimental.
The presence of carbon dioxide may at times have beneficial effects for fish because C02 lowers
the pH On the other hand, lowering the pH would increase the toxicity of cyanides. The sensitivity
of fish to carbon dioxide appears to decrease directly with the increase in temperature.
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Reagents
1. Phenolphthalein indicator solution : 500 mg of phenolphthalein is dissolved in 10 ml ethanol

or methanol and made up to 100 ml with distilled water.
2. 0.05 N Sodium hydroxide solution: 2 grams of Sodium hydroxide is dissolved in two liters of
distilled water.
Principle:
Free carbon dioxide is the amount of carbon dioxide dissolved in water and waste water. The
quantity of free carbon dioxide shows the acidic nature of the sample water caused by carbonic acid,
which is formed by reaction of carbon dioxide and water. This can be estimated by titrating against
sodium hydroxide by using phenolphthalein as indicator. Here appearance of pink colour is the end
point
Procedure
1. 50 ml of unfiltered water sample is taken in a measuring cylinder avoiding agitation

2. 5 drops of phenolphthalein indicator, is added to it.
3. Titrated it against 0.05 N sodium hydroxide solution 0.5m1 at a time, stirring the sample
gently but thoroughly by raising and lowering a glass rod.
4. The end point is noted when a definite pink colour persists for 5 min.
Table
S.N Name of Volume of

Sampl
sample volum readings Run (

content
e down.
V) mg/L
Initia Fina
1 1
1
2
3
4
Calculation
Free carbon dioxide as (C02), mg/l =
volume in ml of 0.05 N Sodium hydroxide rundown X 1000. _

sample in ml
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Results
The free carbondioxide content in sample A - mg/L

The free carbondioxide content in sample A - mg/L
The free carbon dioxide content in sample A - mg/L
Discussion :
Carbon dioxide rarely causes direct toxicity to fish. However, high concentrations lower pond
pH and limit the capacity of fish blood to carry oxygen by lowering blood pH at the gills. At a given
dissolved oxygen concentration(e.g., 2 mg/L, milligrams per liter;
same as parts per million, ppm),fish may suffocate when CO levels are high and appear unaffected
when Co is low. Catfish can tolerate 20 to 30 mg/L CO if accumulation is slow and dissolved .
Experiment No.4
Date:
DETERMINATION OF TUBIDITY OF WATER BY SECCHI'S DISC METHOD
Aim: To study turbidity of water samples by Secchi's Disc method

Principle:
Various characters that control the quality of water are taste, smell, colour, amount of dissolved
nutrients, dissolved 02 and C02, pH and different types of plants and animals and their density.
Turbidity of the water body determines the depth upto which light can penetrate and thus affects the
distribution and photosynthesis of phytoplankton and macrophysics. More turbid the water body less
is the thickness of its phobic zone. In polluted water bodies turbidity is due to:
a. Effluents: A water body which receives domestic sewage, run off from adjacent
agricultural fields and liquid wastes from
nearby small and large industries remains
turbid. Heavy rope to lower disk
b. Planktons: A water body may be turbid due

to very high density of phytoplankton and Wooden Disk
zooplanktons, especially when the water 20 m diamete+ Hook screw,
body is rich in nutrients.
Secchi's Disc method
Preparation of Secchi's Disc:

An iron disc of about 6 inches diameter was taken to
which a weight was attached in the centre on one side and
an iron hook on the other side. Tied a plastic rope of
sufficient length to the hook. Divided the upper surface of
the disc into 4 equal
Dr.C.V.Narasimha murthy .aquaculture practice manual
2016-2017; M.Sc. Zoology Final year 3rd semester
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segments and painted two of these white and the other two segments black in such a way that black
and white segments alternate with each other.
PROCEDURE
1. Visited a nearby aquacultural pond.

2. Slowly immersed the Secchi's disc into water vertically holding the rope tightly in the hand
till the black and white segments of the disc just begin to disappear. On reaching to a
particular depth, the disc becomes completely invisible. Mark the length of the rope when the
disc just disappears.(X Cm)
3. Slowly pulled up the disc and find out the length of the rope where the black and white
segments of the disc just reappear(y cm)
4. Found the mean length of the rope by calculating (x+y)/2
Table
S.No. Name of Length the of the Length the of the Average Turbidity
the distance of disk distance of disk Disappearance (in cm)
location disappearance while reappearance while Distance
dipping in to water dipping in to water
X cm Y cm (X+Y)/2
1
2
3
Results
1. The turbidity of water in Pond A
2. The turbidity of water in Pond B
3. The turbidity of water in Pond C
Discussion
From this experiment it can be concluded that the turbidity is variable in different ponds.
Turbidity is a measure of the amount of suspended material in the water and is a direct assessment of
the penetration of light through the water. Turbidity is caused by living organisms such as algae
(minute plants) and zooplankton (small animals), and by nonliving components such as suspended
clay or silt. The lower the turbidity reading the murkier the water and therefore the more shallow the
light penetration.
A certain amount of turbidity is required in dams and ponds. In very clear water freshwater, crayfish
tend to hide during the daylight hours rather than feed. There is also the risk of
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increased predation from birds etc. However, too high a turbidity can also cause problems. For
example, if a high turbidity is due to clay particles in the water then the light penetration is
reduced. As a result of this, photosynthesis by the algae in the water is reduced and limited to the top
layer of water. This can result in a reduction of the amount of oxygen in the water and, in
particular, deoxygenating of the bottom water layer. The pond may also become thermally
stratified.
On the other hand, if the turbidity is caused by algal blooms in the pond, then this also
affects the oxygen levels in the water. For example, if the bloom is too large, then large amounts of
oxygen are produced during the day by photosynthesis. During the nighttime, the plants respire and
consume oxygen which, under certain conditions, also leads to depletion in the oxygen levels and
therefore possible loss of stock.
Experiment No. 5
Date:
DETERMINATION OF CARBONATES AND BICARBONATES
Aim: To determine the carbonate and bicarbonate content of water.
Introduction:
Alkalinity is important for fish and aquatic life because it protects or buffers against rapid pH
changes. Higher alkalinity levels in surface waters will buffer acid rain and other acid wastes and
prevent pH changes that are harmful to aquatic life. Large amount of alkalinity imparts bitter taste in
water. The principal objection of alkaline water is the reactions that can occur between alkalinity and
certain citations in waters. The resultant precipitate can corrode pipes and other accessories of water
distribution systems. As waters containing excess caustic (hydroxide) alkalinity are not to be
discharged into natural water bodies or sewers.
Alkalinity as carbonate and bicarbonate of saline water is very important in tertiary
recovery processes for recovering petroleum. Alkaline water offers better wetting to the formation
rock and improve oil release. As an additional benefit, ions that provide alkalinity absorb on rock
surfaces occupying adsorption sites and decrease the loss of recovery chemical by adsorption. The
alkalinity value is necessary in the calculation of carbonate scaling tendencies of saline waters. The
alkalinity acts as a pH buffer in coagulation and lime-soda softening of water. In wastewater
treatment, alkalinity is an important parameter in determining the amenability of wastes to the
treatment process and control of processes such as anaerobic digestion, where bicarbonate
alkalinity, total alkalinity, and any fraction contributed by volatile acid salts become
considerations.
Principle:
Carbonate and bicarbonate ions in the sample can be determined by titrating it wit against
standard sulphuric acid (xzsoa) using phenolphthalein and methyl orange as indicators. Addition of
phenolphthalein gives pink red colour in the presence of carbonates and titration with sulphuric acid
converts these carbonates into bicarbonates and decolorize s the red colour as shown
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below:
2 Na2CO3 + H2SO4 NaHCO3 + Na2SO4
Thus the carbonates neutralization is only half way. These carbonates along with the
already present ones are then determined by continuing the titration using methyl orange indicator
which gives yellow colour in presence of bicarbonates. On complete neutralization of bicarbonates
the yellow colour will change to red.
2 NaHCO3 + H2SO4 1'2SO4 + 2 H2O+ 2 CO2
Obviously the bicarbonate titer value will be less if carbonates were not present.
(Absence of pink colour). In such a situation, either the same aliquot is used for bicarbonate
titration or a fresh sample is analyzed for this. If carbonates are present and neutralized, the
volume of H2SO4 used in the first phase (carbonate titration) is to be doubled to get the actual
volume needed for complete neutralization of the carbonates.
Reagents:
1. Saturated H2S04 (0.01N): Carefully added 2.8 ml of conc. H2SO4 to one liter volumetric
flask and diluted to one liter with distilled water, the strength will be approximately 0.1N
H2S04. Diluted 100 ml of this solution to 1 liter to obtain 0.01N H2SO4. Standardized it
against primary standard, Na2CO3
2. Standard Na2CO3 (0.01N): Dissolved 5.3 gm of A.R. Grade Na2CO3 in one liter
volumetric flask with distilled water, the strength will be 0.1N Na2CO3. Dilute 100 ml of
this solution to get 0.01N. this is used for standardization of 0.01N H2SO4.
3. Phenolphthalein (0.25%): Dissolved 25 mg of pure Phenolphthalein powder in 100 ml of

60% ethyl alcohol.
4. Methyl Orange (0.50%): Dissolved 0.5 gm of dry methyl orange powder in 100 ml of
95.0% ethyl alcohol.
Procedure:
1. Transferred 25 ml of water sample into a 150 ml conical flask. Added 2-3 drops of
Phenolphthalein.
2. If pink red colour appears, titrated it against standard H2SO4 till colour disappears. The burette
reading (volume used) is designated as Y ml.
3. To this colorless solution or in original sample (25 ml) added 2- 3 drops of methyl orange. This
will develop the yellow colour.
4. Again Citrate with standard H2SO4 till colour changes from yellow to rosy red. Recorded the
volume of H2SO4 as Z ml. This volume corresponds to initial carbonate changed to
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bicarbonates plus initial bicarbonates present in irrigation water.

5. Run a blank (25 ml distilled water) and subtracted from the titer value to avoid error due to any
impurity of chemicals
Calculations: Carbonates (mill equivalents / liter)

2 (Y - Bl x Normality of H2SO4 x 1000
Sample in ml.
Bicarbonates (mill equivalents / liter)
(Z - B) - 2 (Y- B) x Normality of H2SO4 x 1000
Sample in ml.
Here Y is the burette reading (ml of H2SO4) after phenolphthalein is neutralized and Z is the final
burette reading (total volume of H2SO4) after methyl orange.
S Name of Sam
. the ple Burette readings Volume Volume of
N Sample volu
of sulphuric
o me
sulphuri acid run
c acid down
run (Z)with
down phenolpht
(Y) with halein and
phenolp methyl
hthalein orange
Initial Middle Final
1 Blank 25
ml
2
3
4
Results
S.No Name of the Total Carbonate Content Total Bicarbonate Content
Sample
1
2
3
4
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Discussion:
Alkalinity is the capacity of water to neutralize acids without an increase in pH. This parameter is a
measure of the bases, bicarbonates (HCO3 -), carbonates (CO3-) and, in rare
instances, hydroxide (OH-). Total alkalinity is the sum of the carbonate and bicarbonate
alkalinities. Some waters may contain only bicarbonate alkalinity and no carbonate alkalinity.
The carbonate buffering system is important to the fish farmer regardless of the production
method used. In pond production, where photosynthesis is the primary natural source of oxygen,
carbonates and bicarbonates are storage area for surplus carbon dioxide. By storing carbon dioxide
in the buffering system, it is never a limiting factor that could reduce photosynthesis, and in turn,
reduce oxygen production. Also, by storing carbon dioxide, the buffering system prevents wide daily
pH fluctuations.
Without a buffering system, free carbon dioxide will form large amounts of a weak acid
(carbonic acid) that may potentially decrease the night-time pH level to 4.5. During peak periods of
photosynthesis, most of the free carbon dioxide will be consumed by the phytoplankton and, as a
result, drive the pH levels above 10. As discussed, fish grow within a narrow range of pH values and
either of the above extremes will be lethal to them.
In recalculating systems where photosynthesis is practically non-existent, a good buffering

capacity can prevent excessive buildups of carbon dioxide and lethal decreases in pH. It is
recommended that the fish farmer maintain total alkalinity values of at least 20 ppm for catfish
production. Higher alkalinities of at least 80-100 ppm are suggested for hybrid striped bass. For
water supplies that have naturally low alkalinities, agriculture lime can be added to increase the
buffering capacity of the water.
References
Singh, Dhyan, Chhonkar, P.K. and Pande R.N., 1999, Assessment of Irrigation Water Quality in
"Soil, Plant, Water Analysis" - A methods manual, Indian Agricultural Research Institute, Indian
Council of Agricultural Research, New Delhi, 3 : 76-78.
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Experiment No.6
Date:
DETERMINATION OF ALKALINITY OF WATER SAMPLES
Aim: To determine the Alkalinity of water samples
Introduction:
Alkalinity is important for fish and aquatic life because it protects or buffers against rapid pH
changes. Higher alkalinity levels in surface waters will buffer acid rain and other acid wastes and
prevent pH changes that are harmful to aquatic life. Large amount of alkalinity imparts bitter taste in
water. The principal objection of alkaline water is the reactions that can occur between alkalinity and
certain citations in waters. The resultant precipitate can corrode pipes and other accessories of water
distribution systems. As waters containing excess caustic (hydroxide) alkalinity are not to be
discharged into natural water bodies or sewers.
Alkalinity as carbonate and bicarbonate of saline water is very important in tertiary
recovery processes for recovering petroleum. Alkaline water offers better wetting to the formation
rock and improve oil release. As an additional benefit, ions that provide alkalinity absorb on rock
surfaces occupying adsorption sites and decrease the loss of recovery chemical by adsorption. The
alkalinity value is necessary in the calculation of carbonate scaling tendencies of saline waters. The
alkalinity acts as a pH buffer in coagulation and lime-soda softening of water. In wastewater
treatment, alkalinity is an important parameter in determining the amenability of wastes to the
treatment process and control of processes such as anaerobic digestion, where bicarbonate
alkalinity, total alkalinity, and any fraction contributed by volatile acid salts become
considerations.
Principle:
Carbonate and bicarbonate ions in the sample can be determined by titrating it wit against
standard sulphuric acid (H2S04) using phenolphthalein and methyl orange as indicators. Addition of
phenolphthalein gives pink red colour in the presence of carbonates and titration with sulphuric acid
converts these carbonates into bicarbonates and decolorize s the red colour as shown below:
2 Na2CO3 + H2SO4 ! N a H C O 3 + Na2SO4 s
Thus the carbonates neutralization is only half way. These carbonates along with the
already present ones are then determined by continuing the titration using methyl orange indicator
which gives yellow colour in presence of bicarbonates. On complete neutralization of bicarbonates
the yellow colour will change to red.
2 NaHCO3 + H2SO4 N~22SO4 + 2 H2O + 2 C02
Obviously the bicarbonate titer value will be less if carbonates were not present.
(Absence of pink colour). In such a situation, either the same aliquot is used for bicarbonate
titration or a fresh sample is analyzed for this. If carbonates are present and neutralized, the
volume of H2SO4 used in the first phase (carbonate titration) is to be doubled
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18
to get the actual volume needed for complete neutralization of the carbonates.
Reagents:
1. N/50 H2 SO a : Carefully add 0.7 ml of co nc . H2SO4 in to a bottle and dilute to a 500 ml
with distilled water. From this stock solution took one ml and diluted to 1000 ml. Then the
strength will be approximately n/50 H2SO4.
2. Phenolphthalein indicator : Dissolve 1 gm of pure Phenolphthalein powder in 10 ml of ethyl

alcohol and made up to 100 ml with distilled water
Procedure:
1. Transferred 25 ml of water sample into a 100 ml conical flask. Added 2-3 drops of
Phenolphthalein.
2. If pink red colour appears, titrate it against standard H2SO4 till colour disappears. The
burette reading (volume used) is designated as Y ml.
3. To this colorless solution or in original sample (25 ml) add 2- 3 drops of methyl orange.
This will develop the yellow colour.
4. Again titrate with standard H2SO4 till colour changes from yellow to rosy red. Record the
volume of H2SO4 as Z ml. This volume corresponds to initial carbonate changed to
bicarbonates plus initial bicarbonates present in irrigation water.
5. Run a blank (25 ml distilled water) and subtract from the titer value to avoid error due to any
impurity of chemicals
Calculations:
Phenolphthalein Alkalinity = Y ml X 1000 ml
of sample
Total alkalinity = Z ml X 1000 ml of sample
S. Nam Samp Volume of Volume of

No e of le Burette readings sulphuric sulphuric
the volu acid run acid run
samp me down (Y) down
le with (Z)with
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19
phenolphtha phenolphtha
lein lein and
methyl
orange
Initi Midd Fin

al le al
1
2
3
4
Results
S.No Name of the Phenolphthalein Alkalinity Total alkalinity
Sample
1
2
3
4
Discussions
Alkalinity and hardness are both important components of water quality. However, these two
aspects of water chemistry are commonly confused. The confusion relates to the term used to report
these measures, ppm CaCO3 (same as mg/L). Total alkalinity indicates the quantity of base present
in water -- bicarbonates, carbonates, phosphates, hydroxides, etc. Hardness represents the overall
concentration of divalent salts (calcium, magnesium, iron, etc.) but does not identify which of these
elements is/are the source of hardness. It is important to recognize the difference between hardness
and total alkalinity when farming aquatic animals.
The determination of whether water is acid, neutral or base is defined by pH. However,
alkalinity measures the total amount of base present and indicates a pond's ability to resist large pH
changes, or the "buffering capacity." The most important components of alkalinity are carbonates
and bicarbonates. The total alkalinity concentration should be no lower than 20 mg/L CaCO3 in
production ponds. Pond pH can swing widely during the day, measuring from 6 to 10, when
alkalinity concentrations are below this level. Large daily changes in pH can cause stress, poor
growth and even death of the farmed animals. Most aquatic organisms can live in a broad range of
alkalinity concentrations. The desired total alkalinity level for most aquaculture species lies between
50-150 mg/L CaCO3, but no less than 20 mg/L
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References
Mushy J.D and J.D. Mushy 2006: Fundamentals of freshwater Biology. Narendra Publishing
House. New Delhi. Pp172-173
Singh, Dhyan, Chhonkar, P.K. and Pande R.N., 1999, Assessment of Irrigation Water Quality in
"Soil, Plant, Water Analysis" - A methods manual, Indian Agricultural Research Institute, Indian
Council of Agricultural Research, New Delhi, 3 : 76-78.
Experiment No. 7
Date:
EXPERIMENT ON DETERMINATION OF CALCIUM HARDNESS
AIM: To determine the hardness of water
INTRODUCTION
Water hardness is an expression for the sum of the calcium and magnesium cat-ions
concentration in a water sample. Calcium is usually found in highest concentrations in natural
water. The presence of calcium in water results from deposits of lime stone, gypsum etc. Calcium is
one of the principal cations involved in water hardness. Calcium hardness is the estimation of
hardness due to calcium in water. These cations form insoluble salts with soap and decrease the
cleaning effectiveness of soap. They also form hard water deposits in hot water heaters. The
calcium content may range from zero to several hundred ppm.
ENVIRONMENTAL SIGNIFICANCE
The relative amounts of Calcium hardness, Carbonate and non-Carbonate hardness present in
water are the factors while determining the most economical type of softening process.
Determination of hardness serves as a basis for routine control of softening processes. Hard water
typically contains high concentrations of Ca and Mg cations, which interfere with the use of the
water for many applications
PRINCIPLE
The quantity of calcium in water will be determined by titrating the water sample with a
standard Ethylene Diamine Tetra Acetic acid (EDTA) of known volume and concentration. Based on
the stoichiometry of the reactions and the number of moles of EDTA required to reach the end
point, the concentration of calcium content in water is calculated. An indicator, ammonium
purpurate which combines only with calcium is used. The indicator imparts a pink color to the
solution while there are calcium and magnesium ions that have not complexed with EDTA. Once
the endpoint has been reached and there is no more uncomplexed Ca or Mg, the solution will turn to
purple color. No hint of pink color will be left.
Materials Required
1. Burette
2. Pipettes with elongated tips
3. Conical flask
4. 250 ml Graduated Cylinder
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5. Standard Flask
6. Beakers
7. Wash bottle
CHEMICALS REQUIRED
1. Ammonium Chloride
2. Ammonium Hydroxide
3. EDTA
4. Erichrome Black T
5. Ethanol
1. Standard EDTA Solution (0.02 M) : Weighed 3.723g of EDTA sodium salt and dissolved in
1000 ml distilled water..
2. Ammonia Buffer: for 11.4 ml of ammonium hydroxide , 1.35 ml of ammonium chloride is

added and dissolved in 20 ml distilled water
3. Erichrome Black T solution : 0.5 grams of Erichrome Black t is dissolved in l Oml ethanol
and made upto 100 ml with distilled water.
PROCEDURE:
50 ml of sample water was taken into a conical flask and for that one ml of Ammonical buffer is
added and titrated it against 0.02 M EDTA solution using Erichrome Black T as indicator till the
red colour become blue colour.
CALCULATION
Total hardness as Ca C03(mg/L) = Volume of EDTA X 1000
Volume of Sample
TABLE
S.No Sample Volume of Total Hardness
Name
of the Volume EDTA run as mg
Burette down CaCO3/L
sample readings
Initial Final
1
2
3
4
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Results
DISCUSSION
Calcium and magnesium ions are important contributors to water hardness. When water is heated,
they break down and precipitate out of solution, forming scale. Maximum limits have been
established. Magnesium concentrations more than 100 mg/L may have a laxative effect on some
people.
Experiment No.8
Date:
EXPERIMENT ON DETERMINATION OF RESIDUAL CHLORINE
Aim: To determine residual chlorine in the given water sample
INTRODUCTION
Chlorination is primarily adopted to destroy or deactivate disease-producing microorganisms
in the public water supplies and polluted rivers. Chlorine is usually added to water in gaseous form
or as sodium or calcium hypochlorite. It has been practiced over several years. When chlorine is
added to water, some of the chlorine reacts first with organic materials and metals in the water and is
not available for disinfection (this is called the chlorine demand of the water).
The remaining chlorine concentration after the chlorine demand is accounted for is called
total chlorine. Total chlorine is further divided into: 1) the amount of chlorine that has reacted with
nitrates and is unavailable for disinfection which is called combined chlorine and, 2) the free
chlorine, which is the chlorine available to inactivate disease-causing organisms, and thus a measure
to determine the portability of water The word "residual" means "remainder" or "that which is left",
and as the name suggests the chlorine residual test is used to measure the amount of chlorine
remaining in the water at the time the test is made. The chlorine residual is usually tested in finished
water which is ready to be released into the distribution system, although operators must also ensure
that there is adequate residual at the extreme ends of the distribution system. Although the pros and
cons of disinfection with chlorine have been extensively debated, it remains the most widely used
chemical for disinfection of water. Excess Chlorination may produce adverse effects. Potentially
carcinogenic chloroorganic compounds such as chloroform may be formed. To fulfill the primary
purpose of chlorination and to minimize any adverse effects, it is essential that proper testing
procedures be used. Several methods for measurement of total residual chlorine are available
including iodometric methods, amperometric titration methods, and N,Ndiethyl-pphenylenediamine
(DPD) methods. In this module we are going to learn Iodometric method of residual chlorine
determination.
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Chlorine residuals determination is used to control chlorination of domestic and industrial

wastewaters. Active chlorine (free and combined) should be determined at each stage in the
treatment process of drinking water and in the water mains in order to guarantee bacteriologically
impeccable water. Chlorine determination is important to avoid bad odour and change in the taste of
water. It is determined in the swimming pools to avoid ill effects due to excess chlorination.
Determination of chlorine residual in water distribution is useful to find the source of contamination
or leakage points, so as to supply wholesome water to the consumer. Thus, the main purpose for the
chlorination of water supplies and polluted waters serves primarily to destroy or deactivate
disease-producing micro-organisms.
PRINCIPLE
The starch-iodide titration method, one of the oldest methods for determining chlorine, is very
non-specific for oxidants and generally is used for total chlorine testing at levels above 1 mg/L C12.
Chlorine will liberate free iodine from potassium iodide (KI) solutions at pH 8 or less. The liberated
iodine is titrated with a standard solution of sodium thiosulphate (Na2S2O3) with starch as the
indicator. This method is based on reaction with thiosulfate solution The end point of the titration is
indicated by the disappearance of the blue-colored, starch-iodide complex.
APPARATUS REQUIRED
1. Burette & Burette Stand

2. Porcelain Tile
4. Pipette Bulb
5. Wash Bottle
6. 250 ml Graduated Cylinder
7. Conical Flask (Erlenmeyer flask)
CHEMICALS REQUIRED
1. Acetic Acid, Conc. (glacial)
2. Potassium Iodide, KI, crystals
3. Sodium thiosulphate
4. Starch indicator
5. Distilled or Deionized Water
PREPARATION OF REAGENTS :
1. Sodium Thiosulphate solution (0.O1N) Weighed approximately 2.482 g of sodium

thiosulphate . Transferred to the beaker and dissolved it in boiled distilled water.
Transferred it to the standard flask and make it up to 1000 ml.
2. Glacial Acetic acid
3. Potassium iodide
4. 1% starch solution: dissolved 500 mg of starch in 50 ml distilled water and boiled
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PROCEDURE
1. 200 ml of sample is taken into a conical flask.
2. Added 5 ml Acetic acid. to acidify the sample. It is used to reduce the pH between 3 and 4 in
the conical flask.
3. Added 1 g Potassium Iodide (KI) measured using the spatula and dissolved it by
thoroughly mixing it with stirring rod.
4. Added 1 ml of starch solution and continue the titration until the blue color disappears.
Titrated the solution with standard Na2S2O3 solution until the yellow color of liberated
Iodine is almost faded out. (Pale yellow color)
5. Noted down the burette reading (to know the volume of sodium thiosulphate added).
CALCULATION
Residual (mg/L) = Volume of hypo rundown X normality of hypo X1000X35.5

Chlorine Volume of Sample
TABLE
S.No Name of Sample Volume of Residual
the volume Hypo run chlorine
Burette
sample down content
readings
Initial Final
1
2
3
4
DISCUSSION In the given sample Residual chlorine measured to be = mg/L. It is more than
permissible amount. Active chlorine should be present at each stage of water treatment and
distribution. The residual chlorine at the consumers end should be 0.2 mg/l. Presence of excessive
chlorine gives bad odour and taste and is harmful also. It may lead to cancer, skin and eye
irritation. To avoid the excess chlorination, water sample is to be boiled before the domestic use.
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25
Experiment no. 9
Date:
DETERMINATION OF AMMONICAL NITROGEN IN WATER
Aim: To estimate the Ammonical nitrogen content in water samples
Introduction
Nitrogen compounds are of interest to wastewater treatment plant operators because of the
importance of nitrogen in the life cycles of plants and animals. Nitrogen is a nutrient and occurs
in many forms including ammonia, organic, nitrate and nitrite each of which may be tested for in a
variety of ways. Raw wastewater nitrogen is normally present in the organic nitrogen and ammonia
forms, with small quantities of the nitrite and nitrate forms. Depending on the amount of
nitrification which occurs within the plant, the effluent may contain either ammonia or nitrate
nitrogen. Under normal circumstances, the nitrite form of nitrogen will not be present in large
quantities due to its rapid oxidation or conversion to nitrate.
The presence of large concentrations of ammonia in a stream or lake can create a large
oxygen demand. This demand is caused by the conversion of ammonia to nitrate. High
concentrations of nitrate in wastewater treatment plant effluent can cause algae to grow in large
quantities. Dead and decaying algae can cause oxygen depletion problems which in turn can kill
fish and other aquatic organisms in streams. For this reason, testing for nitrogen in the plant
effluent is critical.
Method: Nessler's Method.
The addition of Nessler reagent to a sample of distillate will produce a color which ranges
from pale yellow to brown depending upon the amount of ammonia present. The pale yellow color
will be present if the ammonia nitrogen level in a 50 ml sample is 20 to 50 micrograms. The
wavelength at which the measurement is made is dependent on the concentration level expected.
EQUIPMENT
1. Spectrophotometer with light path of 1 cm or more or Filter photometer with light path of 1
cm and violet filter with maximum transmittance 400 to 425 nm
2. pH meter
3. Beakers
4. Conical flask
5. Cuvets, matched set
REAGENTS
1. Stock Am m oniu m solu tion ( 1,000 m g /L)

Dissolved 3.819 grams of anhydrous ammonium chloride Cl), (NH4dri ed at 100°C for 1 hour,
in ammonia free water and diluted to a liter.
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2. Standard Ammonium solution (100 mg/L) Dilute d 10.0 ml of
stock solution to 1 L with ammonia free water.
3. Nessler reagent
Dissolved 100 g of Mercury (II) iodide (HgI2) and 70 g of Potassium iodide (KI) in a small
amount of ammonia free water. Caution: Mercuric iodide is toxic. Avoid ingestion. Added this
mixture slowly, with stirring, to a cool solution of 160 g of Sodium hydroxide (NaOH) dissolved
in 500 ml of ammonia free water. Diluted to a liter. Stored this solution in a rubber stoppered
Pyrex bottle in the dark. This reagent may remain stable for up to 1 year.
The reagent should be checked to make sure it yields the characteristic color with 0.1 mg
NH3-N/L within 10 minutes after addition and does not produce a precipitate with small
amounts of ammonia within 2 hours.
PROCEDURE
50 ml of sample is taken into a conical flask. 2 ml Nessler reagent is added to it. Allow 10
minutes for color development. If ammonia nitrogen is extremely low, allowed 30 minute for
colour development. Measured percent transmittance of the sample using a distilled water blank
as reference (%T = 100) at 440 nano meter wavelength.
CALCULATION
Amount of Nitrogen = O.D. of Sample ______ X Amount present standard X 1000 O.D.
of Standard Volume of sample taken
Table
S.No. Name of the Optical density Quantity of Ammonical Nitrogen

Sample
1
2
3
DiscussionWhen feed is eaten by fish it is metabolized into the energy, nutrients, and proteins used for
survival and growth. Ammonia is the principal waste product excreted by fish.
Of all the water quality parameters which affect fish, ammonia is the most important after oxygen,
especially in intensive systems. In small amounts, ammonia causes stress and gill damage. Fish exposed
to low levels of ammonia over time are more susceptible to bacterial infections, have poor growth and
will not tolerate routine handling as well as they should. Ammonia is a killer when present in higher
concentrations. In water, ammonia occurs in two forms, which together are called the Total Ammonia
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Nitrogen, or T . TAN is composed of toxic an-ionized ammonia (NH3) and nontoxic ionized ammonia
(NH4+). Water temperature and pH will effect which form of ammonia is predominant at any given time in
an aquatic system. As ammonia is excreted, it is converted to nitrite (N02-) which is also toxic to fish. This
change from ammonia to nitrite is accomplished by bacteria called Nitrosamines. Another group of
bacteria, called Nitrobacteria, convert nitrite to nitrate (N03+) which is not toxic to fish. Nitrate is used by
plants, including algae, for food. This constant change from ammonia to nitrite to nitrate is called the
nitrogen cycle. In ponds this process takes place in the surface layers of the mud, but tanks or aquaria have
to be provided with biofilters where bacteria can live and flourish.
Experiment No.10
Date:
POND CONSTRUCTION
Aim: To study the construction of pond
Introduction:
Before constructing the pond, land is surveyed to fmd out determine its topography. Marking the area of
proposed pond is the first step in the construction of a fish/prawn pond.
The natural slope where the main wall is to be built should be ascertained. The main wall should be marked
off at the lower end of the pond, where the slope is the greatest.
Designing
The first step while designing fish ponds should be to study the soil type, topography and water
supply. In designing the fish farm, it should be decided as to where and how many nursery, rearing and
stocking ponds are to be constructed. In case of a fish farm constructed solely for the purpose of seed
production, only nursery and rearing ponds may be constructed, with a nominal area for the brood stock
ponds. In case of grow-out farm, more stocking ponds will be constructed to produce table size fish after
stocking fingerlings.
For a composite fish farm all three types of ponds are required and their number should be based on
the intended stocking density.
Fish ponds should be at least one surface acre in size. Ponds smaller than one acre seldom support a
satisfactory fish population over many years. They usually require much more intensive fish management and
may not justify the costs.
It is important to know the exact size, maximum depth, average depth, and water volume of the
pond. This information becomes useful in calculating the amount of herbicide needed for weed control and
the number of fish fingerlings needed for stocking.
Different lands of pond
Freshwater fish ponds differ according to their source of water, the way in which water can be
drained from the pond, the material and method used for construction and the method of use for fish
farming. Their characteristics are usually defined by the features of the landscape in which they are built.
Ponds can be described as follows.
According to the water source
1. Ponds can be fed by groundwater:
(a) Spring-water ponds are supplied from a spring either in the pond or very close to it. The water
supply may vary throughout the year but the quality of the water is usually constant.
(b) Seepage ponds are supplied from the water-table by seepage into the pond. The water level in the
pond will vary with the level of the water-table.
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2. Rain-fed ponds:
Rain-fed ponds are supplied from rainfall and surface runoff. No water is supplied during the dry season.
These ponds are often small depressions in impermeable soil, with a dike built at the lower side to retain
more water.
3. Ponds can be fed from a water body such as a stream, a lake, a reservoir or an irrigation canal.
These may be fed directly (e.g. barrage ponds), by water running straight out from the water body to the
ponds, or indirectly (e.g. diversion ponds), by water entering a channel from which controlled amounts can
be fed to the ponds.
4. Pump fed ponds are normally higher than the water level and can be supplied from a well, spring, lake,
reservoir or irrigation canal, by pumping.
According to the means of drainage
1. Undrainable ponds cannot be drained by gravity. They are generally fed by groundwater and/or surface
runoff, and their water level may vary seasonally. Such ponds have two main origins.
2. Drainable ponds are set higher than the level to which the water is drained and can easily be drained by
gravity*. They are generally fed by surface water such as runoff*, a spring or stream, or are pump-fed.
3. Pump-drained ponds may be drainable by gravity to a certain level, and then the water has to be pumped
out. Other ponds, similar to undrainable ponds, must be pumped out completely. These ponds are only used
where groundwater does not seep back in to any extent.
According to the construction materials
1. Earthen ponds are entirely constructed from soil materials. They are the most common, and you will
learn primarily about these ponds in this manual.
2. Walled ponds are usually surrounded by blocks, brick or concrete walls. Sometimes wooden planking or
corrugated metal is used.
3. Lined ponds are earthen ponds lined with an impervious material such as a plastic or rubber sheet.
According to the construction method
1. Dug-out ponds are constructed by excavating soil from an area to form a hole which is then filled with
water. They are usually undrainable and fed by rainfall, surface runoff or groundwater.
2. Embankment ponds are formed without excavation by building one or more dikes above ground level to
impound water. They are usually drainable and fed by gravity flow of water or by pumping.
3. Cut and-fill ponds are built by a mix of excavation and embankment on sloping ground. They are
usually drainable, and water, which is impounded within the dikes, is fed by gravity or by pumping.
According to the use of the pond
1. Spawning ponds for the production of eggs and small fry;
2. Nursery ponds for the production of larger juveniles;
3. Brood ponds for broodstocks rearing;
4. Storage ponds for holding fish temporarily, often prior to marketing;
5. Fattening ponds, for the production of food fish;
6. Integrated ponds which have crops, animals or other fish ponds around them to supply waste
materials to the pond as feed or fertilizer;
Three basic pond types
Ponds can be conveniently grouped into three basic types depending on the way the pond fits in with the
features of the local landscape.
SUNKEN POND:
1. The pond floor is generally below the level of the surrounding land.
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29
2. The pond is directly fed by groundwater, rainfall and/or surface runoff It can be but is not normally
supplemented by pumping.
3. The sunken pond is undrainable or only partially drainable, having been built either as a DUG-OUT
POND or to make use of an EXISTING HOLLOW or DEPRESSION in the ground, sometimes with
ADDITIONAL EMBANKMENTS to increase depth.
BARRAGE POND:
1. They are created in the bottom of a valley by building a DAM across the lower end of the valley. They
may be built in a series down the valley.
2. The barrage pond is drainage through the old river bed.
3. If large floods are present, the excess water is normally diverted around one side of the pond to keep the
level in the pond constant. A DIVERSION CANAL is built for this purpose; the pond water supply is then
controlled through a structure called the WATER INTAKE.
4. Directly fed from a nearby spring, stream or reservoir, the water enters the pond at a point called the
INLET and it flows out at a point called the outlet.
5. To protect the dike from floods, a SPILLWAY should be built.
DIVERSION POND:
1. The diversion pond is fed indirectly by gravity or by pumping through a diversion canal (which becomes the
MAN FEEDER CANAL), from a spring, stream, lake or reservoir. The water flow is controlled through a
water intake. There is an inlet and an outlet for each pond.
2. The diversion pond can be constructed:
either on sloping ground as a cut-and-fill pond;
or on flat ground as a four-dike embankment pond sometimes called a PADDY POND.
3. It is usually drainable through a drainage canal.
The walls should always be at least 30 cm higher than the water level for a small pond, and at least 50 cm
The pond bottom should usually have a slope of 2-5%.

Parts of a 14s i pone! {
higher for a larger pond.
The most important feature is to have the pond bottom slope such that the pond can be drained.
If the pond site has a natural slope, the dyke or main wall should be constructed at the low level side.
30
semester
31
CRsi
When the pond walls are constructed, the excavated soil can be placed on the top and planted with grass.
This fertile top soil will root grass easily and this will help keep the walls from eroding.
The pond bottom must be cleared by removing small rocks, roots, and stumps to prevent the nets from
getting caught and torn during harvesting.
If grass is found in the pond bottom, it need not be removed, because after filling up the pond with water
the grass will die and add nutrients to the water.
When the stakes have been established for construction of dykes, about 2' top soil should be
removed as it consists of large amounts of roots and other organic material.
The core trench is cut immediately after the removal of the top soil. If the soil is porous, the seepage
problem may arise at a later stage. It would be essential to provide a clay core in order to prevent seepage.
A soil which is a mixture of sand and clay is best.
Pure clay soil will crack and leak.
If pure clay is to be used, it must be mixed with other soil before it can be used.
Turf, humus or peaty soils should not be used.
All stones, wood pieces and other material which may rot or weaken the wall must be removed before
building begins.
Construction of dyke
Construction of earthen dyke is always economical.
Soil obtained from digging can
be used to
prepare the
earthen dyke.
32
semester
33
Construction of dyke
The filling of earth should be done in layers not exceeding 20 cm in height and consolidate each
layer by watering and ramming.
The earth work for the dykes should be thoroughly compacted so that even minor seepage can be checked. If
the fish farmer is economically sound, he can go for stone pitched dykes. The dykes of a pond should be
strong enough to withstand weather action. In big ponds erosion of dykes is a problem which requires regular
attention. Brick or stone pitching may be provided to arrest erosion of dykes. Earthen dykes can be protected
from erosions with bamboo piling. Holes should be closed immediately with stiff clay mixed with lime and
cementing material and should be compacted properly. By using concrete blocks, stones or bricks the earthen
dykes will be protected more permanently from crab or rat holes. Side slopes of embankments depend upon
the nature of material used for construction. The slopes should be flatter than the angle. Soil with a lot of clay
in it can have a greater slope on the outside wall than on the inside wall. A typical embankment is built with
an outside slope of 1:1 and an inside slope of 1:2. A slope of 1:2 means that for every increase in 2m width
there is a change of 1 m in height. Once the embankment is constructed, it is better to plant grass on it. The
grass roots help to hold the wall together and prevent erosion of the soil.
Crass Goaa I I Q a c a uter cz, er
P e t
,- ~. tares--r ±
Drainage system
A drainage system is used to empty the pond.
It consists of the outlet system for letting water out of the pond and the drainage ditches which carry the
water away from the pond.
semester
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Drainage of the pond water

The best and easiest way to have a good drainage system is to build the pond in a place which
provides a good slope.
The drainage system must be built before the pond embankment because some drainage devices go through the
walls.
One of the easiest ways to drain the pond is to place a bamboo or plastic pipe through the base of the wall
into the middle of the pond.
The end of the pipe, which is inside the pond, should have a screen over it to keep fish from entering the
pipe. The other end of the pipe is plugged with wood or clay. To drain the pond during harvest time, the
plug is pulled out.
Other methods of draining the ponds are the siphon and the pump.
Drainage of the pond

Sluice
The sluice can be a screened gate in a water channel going into the pond or drainage gate leading
water out of the pond.
The sluice can be made of wood, cement and brick. It can be made up of one or two wooden gates
which are removed to empty or fill the pond.
A sluice also has a screen gate to keep unwanted fish from entering at the inlet and pond fish from
leaving at the outlet.
Water inlet
All the ponds, except for those filled directly by a spring or by rainwater, need water inlets. During the
construction of inlets, filters should be used in the channel so that the unwanted fish or other materials do
not enter into the pond and the water is clean.
A water inlet can be as simple as a bamboo pipe of good diameter running from a water source through the
wall into the pond.
The inlet pipe should be placed above the water level.
A wire screen makes a good filter.
The horizontal screen is very effective.
A nylon mesh bag makes a good filter and can be fixed to the inlet pipe.
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A sand and gravel filter is also used, but it requires a small tank at the water inlet, it is more effective and
economical.
If the water is muddy, or has plenty of leaves or grass in it, the wire screen is better.
If the water source is free from organic material, the mesh bag will work.
If the water contains unwanted fish and more organic matter, sand and gravel filters are best.
Sealing the pond bottom
Leaking Ponds
One of the most common farm pond problems is heavy water loss through leakage. The ability of the pond to
retain water depends largely on the characteristics of the soil at the pond site. Most leaky pond problems can
and should be prevented by cautious site selection. Before building a pond, be sure to test the capacity of the
soil to hold water. Soils with a high clay content will minimize seepage since clay particles tend to
swell when wet and, thereby, provide a good bottom seal.
If the soil has more clay in it, no special sealing is needed.
If the bottom is sandy, it should be sealed to hold the water. To seal the bottom a clay core lining is built
over the pond bottom.
Another method of sealing the pond bottom is with cement blocks, but it is expensive.
The most commonly used pond sealant is betonies clay. Mennonite is most effective on sandy soils that
contain insufficient amounts of clay. For best results, betonies should be spread evenly over the dry pond
bottom at a rate of 50 lbs/100 ft (20,000 lbs/acre) mixed with the existing soil, moistened, and then
compacted with a roller.
Sealing with flexible plastic sheeting of polyethylene, or plastic or vinyl, or butyl or rubber sheet liner at
least 2 mm thick is another method of sealing.
METHOD:
Few ponds were surveyed in the Ramayapatnam village of Gudlur Mandal of Praksam district.
Data collection
Date of survey = 6-8-16

Location = adjacent to Buckingham canal,
Type of soil = sandy cum loamy
Extent = 1.5 acre
Water source = Ground water and Buckingham canal
Purpose of pond = prawn rearing
Pond measurements =
Area = 65340 sq feet. =

Length 400 feet = 163
Width feet = 8 feet = 3
Height of bond feet
Berm length = drainable pond
Drain Type = dug out pond =
barrage pond
Slope =2-3%
Inner slope =1:2
Out slope = 1: 1.5
Dr.C.V.Narasimha
semester
murthy .aquaculture practice manual 2016-2017; M.Sc. Zoology Final year 3rd
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Dyke depth = 8 feet

Water depth = 5 ft
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` Cr ass
G r oe s umac c r r Dar e r
Drainage system
A drainage system is used to empty the pond.
It consists of the outlet system for letting water out of the pond and the drainage ditches which carry the
water away from the pond.
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Experiment No 11.
Date:
SOIL QUALITY ANALYSIS OF POND

AIM: To determine the soil quality of aquaculture pond
INTRODUCTION
Aquaculture ponds are normally built of soils. Properties of soils should be considered in selecting a
site, designing earthwork, and specifying construction methods to provide a water-tight pond with stable
levels and bottom slopes. Interactions between soil and water that influence water quality in ponds must not
be ignored, because poor soil condition in ponds can impair survival and growth of aquaculture species. For
example, acidic soils can cause low pH and total alkalinity in ponds, and unless lime is applied, ponds may be
unsuitable for aquaculture. A large amount of information is needed for proper planning, design and
construction of aquaculture ponds. Sometimes lack of attention to soil properties result in aquaculture ponds,
which cannot be used to their full potential.
The productivity of a pond depends upon its soil -water characteristics. Generally, fish shrimp
production is low in ponds located in agriculturally poor productive soils and high in those placed in fertile
soils. A satisfactory pond soil is the one in which mineralization of organic matter takes place rapidly and
nutrients are absorbed, held and released slowly over a long period. Further, in bottom soil, a series of
chemical and biochemical reactions take place resulting in either the release of nutrients from soil to water or
absorption of nutrients from water by the soil and microbial population. This process governs the growth and
population of the micro and macro food organisms in the fish/shrimp ponds. To understand the complete
pond ecosystem, it is essential to study the characteristics of pond soil and water to increase the productivity
of the ponds in general and thereby augmenting fish and shrimp production.
Shrimp farms have been constructed on variety of coastal lands -intertidal fallow land, dry and
saline fallow land, unproductive and marginal agricultural land, and to a lesser extent in wetlands like
marshes and mangroves. Coastal areas of the country have vast resources of saline affected fallow lands,
which cannot be used for any productive purpose. Presently, about 10.0 % of the resources available are
being utilized for shrimp aquaculture. Shrimp aquaculture requires natural resources like land, water, and
biological resources like seed and feed
Material
Dr.C.V.Narasimha murthy .aquaculture practice manual 2 0 1 6 - 2 0 1 7 ; M.Sc. Zoology Final year 3 r d

semester
39
1. ELICO Water and soil quality analyzer ( Field Kit)
ELICO PE-138, Microprocessor based Multi Parameter Water

Quality Analyzer. Measures directly pH, EMF, Temperature,
Electrolyte Conductivity (EC), Total Dissolved Solids (TDS),
Dissolve Oxygen (DO) & Salinity, Colorimetric, Abs./%T and
Turbidity in Water Sample one at a time.
SALIENT FEATURES
1. Light weight, Portable and Easy to carry on field
2. Unique DO measurement in Air saturated waters using
Conventional Polarographic Probe.
3. Programmable Calibration upto 3 points for pH with
Memory Back-up.
4. Useful for Potentiometric & Redox measurements.
5. Automatic / Manual Temperature Compensation.
6. Accurate & Highly Stable.
7. Elegantly styled into a Briefcase for operational
convenience in Field
8. Operates on Rechargeable Battery & Mains Supply.
Method:
10 grams of soil was taken from the pond floor and dissolved in 100 ml distilled water and
analyzed the following parameters by using different probes.
Texture:
Texture refers to the relative proportions of particles of various sizes such as sand, silt and clay
in the soil. The percentages of sand, silt and clay in a soil could be determined in a soil in the field, soil
texture could be estimated by the following methods
Feel method:
In this method, the soil is moistened with water and rubbed between the thumb and fingers. The
way the wet soil "slicks out" gives a good idea of the clay content. The sand particles are gritty, the silt
has a floury or talcum - powder fell when dry and is only moderately plastic and sticky when wet.
Accuracy of this method depends largely on experience.
Results.
S.No. Parameter Result
1 Texture Sandy -clay

2 PH 6.9
3 Electrolyte conductivity 45 mS
5 Salinity 12 ppt
6 Total dissolved solids 70 ppt
7 Turbidity 45 NTU
Dr.C.V.Narasimha murthy .aquaculture practice manual 2 0 1 6 - 2 0 1 7 ; M.Sc. Zoology Final year 3 r d
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By using different probes that are available in the water and soil analyzer kit various parameters were
determined.
Discussion
Soil texture
The nature and its properties of the parent material determine the soil texture. An ideal pond
soil should not be too sandy to allow leaching of the nutrients or should not be too clayey to keep all
the nutrients absorbed on to it.
Cation exchange capacity
It is the total quality of cations which a soil can absorb by cation exchange is termed as c e c
of the soil. Expressed as mille equivalents / 100 gm soil (meq / 100 gm of soil ). Higher c e c
indicates more concentration of easily exchangeable. The cations are called exchangeable bases.
Ca++, Mg++, Na+ and K+ ions.
Water holding capacity
Capacity of soil to hold water. Import soil character. Clay-silt soils are best for fish ponds. Soil
porosity, particle density, bulk density. Import physical qualities of pond soils. which help to
aeration, filtration, percolation, adsorption of nutrients etc?
Acid sulphate soils
Acid sulphate soils from mine spills and coastal mangroves contain high levels phyrite (fe82
1-6%). Sediments containing pyrites on oxidation results in sulfuric acid. Sulfuric acid reduces the ph
of water when pond is filled. Acid sulphate soils usually originate is pond dykes. Soil acidity
The ideal range of for soil ph is 6.0-8.0. Water passing over acid soil tends to be a acidic with
low alkalinity and hardness. High concentration of metal ions particularly aluminum (A1+++) and
iron (F e + ) to soil acidity. Acid ponds do not respond well to fertilization.
Bottom soil oxidation
Aeration and water circulation are beneficial in improving bottom soil oxygenation but the
surface layer of soil may still become anaerobic in intensive fish culture ponds due to settlement of
suspended particulate matter.
Soil reaction (pH)
The ph of the soil is one of the most important factors for maintaining pond productivity since
it controls most of the chemical reaction in the pond environment. Near neutral or slightly alkaline ph
is considered to be ideal for fish production. If the ph is too low (strongly acidic). This can reduce the
availability of key nutrients in the water and lower the pond productivity.
Organic carbon content
Organic carbon acts as the source of energy for bacteria and other microbes that release
nutrients through various biochemical processes. Pond soils with less than 0.5% organic carbon is
considered unproductive while those on the range of 0.5-1.5% and 1.5-2.5% to have medium and
high productivity, respectively organic carbon content of more than 2.5% may not be suitable for fish
pond.
Carbon to nitrogen ratio
The c: n ratio of soil influences the activity of soil microbes. This in turn affects the rate of
release of nutrients from decomposing organic matter. The rate of break/down (minerals ratio) is very
fast, and slow at c:n ratio in the range of less than 10, 10-20 and more 20 respectively. In general, soil
c:n ratios between 10 and 15 are considered favorable for aquaculture and ratio of 20:1 or narrower
gives good results.
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Nutrients
Nitrogen, phosphorous and potassium are the major nutrients required by phytoplankton pond
soil with 30 ppm, 30-60 ppm, 60-120 ppm and more than 120 ppm available phosphate (p2o5) are high
productivity. Respectively ponds with less than 250 ppm available soil nitrogen are considered to have
low productivity while concentration is the range 250 to 500 ppm and above 500 ppm are considered to
be medium and highly productive respectively. Generally relatively small amounts of potassium are
needed in fish ponds.
The best method for preventing soils and water quality problems is aquaculture ponds is to
select a site with good soil and an adequate supply of high quality water. If this is done, liming,
fertilization and aeration can prevent most soil and water quality imbalances.
Proper pond management viz. Liming, fertilization, aeration, water exchange and bottom soils
drying and oxidation are the keys to improve soil and water quality in ponds.
Experiment No.12
Date:
WATER QUALITY ANALYSIS OF POND

AIM: To determine the water quality of aquaculture pond
INTRODUCTION
Water quality is a critical factor when culturing any aquatic organism. Optimal water quality varies by
species and must be monitored to ensure growth and survival. The quality of the water in the production
systems can significantly affect the organism's health and the costs associated with getting a product to the
market. Water quality parameters that are commonly monitored in the aquaculture industry include
temperature, dissolved oxygen, pH, alkalinity, hardness, ammonia, and nitrites. Depending on the culture
system, carbon dioxide, chlorides, and salinity may also be monitored. Some parameters such as alkalinity
and hardness are fairly stable, but others like dissolved oxygen and pH fluctuate daily. It is important to
establish a standardized water quality testing protocol for your particular situation. Know the tolerance range
for your culture species, establish critical levels, and be prepared to act if a problem occurs. The chart
below indicates the water quality preferences for some commonly cultured species. For more information
about a particular parameter, click one of the links below. If you need assistance in this area, contact your
local Cooperative Extension office.
Material
2. ELICO Water and soil quality analyzer ( Field
Kit)
ELICO PE-138, Microprocessor based Multi Parameter Water

Quality Analyzer. Measures directly pH, EMF, Temperature,
Electrolyte Conductivity (EC), Total Dissolved Solids (TDS),
Dissolve Oxygen (DO) & Salinity, Colorimetric, Abs./%T and
Turbidity in Water Sample one at a time.
Dr.C.V.Narasimha murthy .aquaculture practice manual

2016-2017; M.Sc. Zoology Final year 3rd semester
42
SALIENT FEATURES
Light weight, Portable and Easy to carry on field
Unique DO measurement in Air saturated waters using Conventional Polarographic Probe.
Programmable Calibration upto 3 points for pH with Memory Back-up.
Useful for Potentiometric & Redox measurements.
Automatic / Manual Temperature Compensation.
Accurate & Highly Stable.
Elegantly styled into a Briefcase for operational convenience in Field
Operates on Rechargeable Battery & Mains Supply.
Method:
By using the different probes that are present in the water quality analyzer at the Ramayapaqtnam village in
an Aquacultural pond different water quality parameters were tested.
Results. The data is presented in the table.
S.No. Parameter Result
1 Dissolved oxygen 5PPM

2 pH 6.9
3 Electrolyte conductivity 45 mS
5 Salinity 12 ppt
6 Total dissolved solids 70 ppt
7 Turbidity 45 NTU
Discussion
Water quality is the most important factor affecting prawn/shrimp health and performance in
aquaculture production systems. Good water quality refers to what the prawn/shrimp wants and not
what we think the prawn/shrimp wants. This means that we must understand the water quality
requirements of the prawn/shrimp under culture very well. Prawn/shrimp live and are totally
dependent on the water they live in for all their needs.
Different prawn/shrimp species have different and specific range of water quality aspects
(temperature, pH, oxygen concentration, salinity, hardness, etc.) within which they can survive,
grow and reproduce.
Within these tolerance limits, each species has its own optimum range, that is, the range
within which it performs best. It is therefore very important for prawn/shrimp producers to ensure
that the physical and chemical conditions of the water remain, as much as possible, within the
optimum range of the prawn/shrimp under culture all the time. Outside these optimum ranges,
prawn/shrimp will exhibit poor growth, erratic behavior, and disease symptoms or parasite
infestations. Under extreme cases, or where the poor conditions remain for prolonged periods of
time, prawn/shrimp mortality may occur. Pond water contains two major groups of substances:
1. Suspended particles made of non-living particles and very small plants and animals, the
plankton.
2. Dissolved substances made of gases, minerals and organic compounds.
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The composition of pond water changes continuously, depending on climatic and seasonal
changes, and on how a pond is used. It is the aim of good management to control the composition to
yield the best conditions for the prawn/shrimp. For producers to be able to maintain ideal pond water
quality conditions, they must understand the physical and chemical components contributing to good
or bad water quality.
Physical Aspects of Water Quality
Temperature
Prawn/shrimp are "cold-blooded" and therefore assume the temperature of the water they live
in. Water temperature is therefore the most important physical factor for prawn/shrimp survival and
growth. Body temperature, and thus the water temperature, has an effect on level of activity,
behavior, feeding, growth, and reproduction of the prawn/shrimp. Each species has its tolerance
limits and optimum range. When water temperatures are outside the optimum range, prawn/shrimp
body temperature will either be too high or too low and prawn/shrimp growth will be affected or the
prawn/shrimp will even die.
Turbidity
Fine solid particles suspended lead to a turbidity. Turbid Water can be said to be "cloudy".
Turbidity can result from suspended solids (clay) or plankton. Clay turbidity in pond water (muddy
water) can be harmful to prawn/shrimp and limit pond productivity. Clay turbidity in pond can be
controlled.
Soil pH and Acidity

Pond water may be acidic, alkaline or neutral. Depending on this, water will react in
different ways with substances dissolved in it. It will also affect in different ways the plants and
animals living in the water. The measure of the alkalinity or acidity of water is expressed by its pH
value. The pH value ranges from 0 to 14, with pH 7 indicating that the water is neutral. Values
smaller than 7 indicate acidity and greater than 7, alkalinity. Prawn/shrimp production can be
greatly affected by excessively low or high pH.
Extreme pH values can even kill your prawn/shrimp. The growth of natural food organisms
may also be greatly reduced. The critical pH values vary according to the prawn/shrimp species, the
size of individual prawn/shrimp and other environmental conditions. For example, prawn/shrimp are
more susceptible to extreme pH during their reproductive seasons, and eggs and juveniles are more
sensitive than adults. Waters ranging in pH from 6.5 to 8.5 (at sunrise) are generally the most suitable
for pond prawn/shrimp production. Most cultured prawn/shrimp will die in waters with pH below 4.5
and 10 or above. Prawn/shrimp reproduction and general performance can be greatly affected at pH
below 6.5 and above 8.5.
How to correct the pH of your pond water

Pond water with pH unfavorable for prawn/shrimp production can be corrected by:
If the pH is below 6.5 (at sunrise), use lime and alkaline fertilizers
If the pH is above 8.5 at sunrise, you can use acid fertilizers
Ensuring that soil pH and acidity are within acceptable limits is a necessary part of managing the
alkalinity, hardness, and pH of the water, which were discussed above. The key is to keep soil pH at
6.5 or above, which will usually maintain water pH, hardness, and alkalinity at desirable levels.
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How to keep soil pH at the right level

1. Drying the pond for at least two weeks after each harvest before refilling and restocking.
2. Applying lime (preferably agricultural limestone) to the pond after each harvest. Normally
lime should be applied to the pond bottom before it is refilled, but if necessary, it can be
applied to the water surface after filling the pond. Only recommended liming materials and
application rates should be used.
3. Pond water pH varies over the course of a 24-hour day. This variation is related to the light
intensity which is important in photosynthetic activity of phytoplankton.
4. pH is lowest at sunrise and as photosynthesis increases as the light intensity increases, more
and more carbon dioxide is removed from the water by the plants causing the pH to increase
5. A peak value is reached in late afternoon.
6. As the light intensity starts decreasing, which reduces photosynthesis less and less carbon
dioxide is removed from the water; as respiration adds more carbon dioxide to the water, pH
starts to decrease.
7. At sunset, photosynthesis stops, but respiration continues for the rest of the night. More and
more carbon dioxide is produced, and pH keeps decreasing until sunrise, when it reaches its
minimum.
Dissolved oxygen in prawn/shrimp ponds

The most important gas dissolved in water is oxygen. Dissolved oxygen (DO) is
essential for respiration and decomposition.
Dissolved oxygen in water comes from atmospheric oxygen and photosynthesis. The
atmospheric oxygen diffuses and dissolves into the water. But the diffusion and its
subsequent dissolves into water is a slow process. The major source of dissolved oxygen in
ponds is photosynthesis. However this process depends on the amount of light available to
the aquatic plants in water (Phytoplankton). Therefore:
1. Oxygen production decreases during cloudy days

2. It stops at night
3. It decreases in increase in water depth the rate of the decrease depending on the water
turbidity
4. How to measure Dissolved Oxygen (DO)
5. DO can be measured by chemical or by electrical methods. Chemical methods rely on
the use of kits which can be bought from shops dealing with laboratory equipment.
They contain chemicals and equipment necessary to determine the DO content with
sufficient accuracy for pond management purposes.
6. Electrical methods use an oxygen meter, this too can be bought from laboratory
equipment shops but it is expensive. Using this equipment, DO can be measured
directly from the pond at any depth. DO and water temperature should be measured at
the same time so as to be able to relate the DO to the temperature. DO is expressed as
mg of oxygen/liter of water (mg/1).
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7. Dissolved oxygen (DO) requirements commonly farmed prawn/shrimps
Fluctuating oxygen level
1. From sunrise to sunset

2.
3. Photosynthesis increases the DO level
4. DO production is higher on clear sky days than on cloudy days
5. The higher the phytop lankton population, the higher the DO production.
6. At night
7. Photosynthesis does not take place
Respiration and decomposition which are the main activities taking place, reduces the DO
8. content until sunrise
9. The higher the plankton population and dead matter, the faster the DO will fall There
may be very little oxygen left by morning and prawn/shrimp may suffocate if corrective
measures are not taken. In over fertilized ponds, where there is very high plankton
density and high turbidity, the DO content of the bottom water may become anoxic
(without oxygen) even during the day. The prawn/shrimp will concentrate at the surface
of the pond to survive. This will be much worse at night.
Where DO test equipments are not available, signs indicating reduced DO in pond water include:
1. Prawn/shrimp not feeding well or even stopping feeding
2. Prawn/shrimp coming to the water surface to breathe from the better oxygenated surface
water
3. The DO content of pond water can be increased in several ways:
4. Through design and management
5. Through structures that cause water to splash e.g. by use of cascades along the inlet canal
and raised inlet pipes before the water gets into the ponds
6. By use of mechanical aerators for the emergency aeration of pond water
7. A simple way to ensure a good supply of atmospheric oxygen to prawn/shrimp ponds is in
the design of the pond. The ponds should be designed such that they take maximum
advantage of the winds. The ponds should be designed so that the lengths are parallel to the
direction of the prevailing winds.
8. Proper pond management can also improve the DO content of the water. The following
measures can be taken before any emergency happens:
9. Flashing the pond by removing the less oxygenated bottom water and replacing it with
better oxygenated water
10. Use of water aerators e.g. mushroom blowers and paddle wheels
Alkalinity and Hardness

It is desirable to maintain both alkalinity and hardness at 40 - 70 mg Calcium carbonate per
liter. This can be done by:
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1. Where water is 'soft' or acidic and soils are acid, apply lime (agricultural limestone) to the
pond soil at recommended rates before to filling the pond
2. Lime may also be added after filling by spreading it uniformly over the water surface.
3. In areas where soils are alkaline and hardness and alkalinity are high, application of lime is
not required.
4. Note that proper management of hardness and alkalinity will usually eliminate the need to
worry about pH.
Ammonia
Un-ionized ammonia (NH3) concentrations in pond water should be kept below 0.5 mg/1
Concentrations of this form of ammonia, which is toxic to prawn/shrimp, are influenced by DO,
pH, and alkalinity; therefore it is important to manage this by:
1. Maintain water alkalinity at 40 mg Calcium carbonate per liter or above
2. Keeping pH near neutral, and at least below 9.0
3. Keeping DO concentrations high
Toxic Materials
Substances toxic to prawn/shrimp and other organisms (herbicides, insecticides, and other chemicals)
should be kept out of the ponds. Ponds should be protected by:
1. Not using insecticides, herbicides, or other chemicals (except for recommended inorganic
fertilizers) in or near your pond
2. Keeping agricultural runoff from the ponds
3. Avoiding spraying agricultural crops near ponds on windy days
Experiment No.13
Date:
ANALYSIS OF PLANKTON
Aim: To analyze living organisms in water samples
Principle:
The productivity and the trophic status of a water body is determined by assessing the number
and type of organisms (micro as well as macro) present in the water body. Water body with very high
density of phytoplankton per unit area is a productive water body. Such water bodies are usually
turbid and have high amounts of nutrients and dissolved oxygen. These water bodies support fairly
large number of organisms of different trophic levels. This is in contrast to non-productive water
bodies, which have very low density of organisms per unit area, fairly transparent waters with low
mineral concentration and dissolved oxygen and also fewer trophic levels. The status of health of a
water body can be determined by analyzing water samples for the number and type of organisms
present in it at a given time. Such assays also help us to find out whether a water body is polluted as
some of the organisms are strong indicators of water pollution.
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Requirement:
Water samples from different water bodies (lake, pond, river etc.), beakers, a few vials or
small test tubes, slides and cover slips, watch glasses, dropper, compound microscope and 5% FAA
(Formalin Aceto Alcohol5:5:90:Formalin: Acetic acid: Ethanol) as preservative
Procedure
1. a liter of water sample is collected from nearby water body (pond lake, reservoir , river
etc).
2. 5 ml of FAA is added to fix and preserve the living organisms present in each sample at the
place of collect ion.
3. Labeled each water sample to indicate the site from which the water sample has been
collected.
4. Left the water samples undisturbed for 48-72 hours.
5. Decanted the clear water, leaving concentrated sediment at the bottom.
6. Transferred the sediment into a vial or a small test tube. Corked and labeled each vial for
future use.
7. With the help of a dropper, transferred a few drops of sediment liquid from a vial into a
watch glass. Diluted the sediment with water if the sediment is highly concentrated.
8. Identified the collected plankton under microscopes
Results
There are xxx number of organism present in the collected samples. Their diagrams and
characteristic features were studied.
Discussion
In Aquaculture ponds, the microscopic plankton serves as live food for shrimp, prawn and
fish. Ecologically, plankton plays a significant role in increasing the phosphorus, potassium and
other nutrients. It also adds organic matter to the pond soil which may improve soil structure,
aeration, soil moisture-holding capacity and water infiltration.
Humic substances in the Pond are excellent natural and organic way to provide a
concentrated dose of essential critical nutrients, vitamins and trace elements not only to stabilize
the plankton blooms in aquaculture ponds but also to improve water quality. The organic matter
(humic substance), present in the Pond , is formed through the chemical and biological
humification of plant and animal matter and through the biological activities of microorganisms
and is having high carbon and nitrogen contents.
The humic contents of Pond i.e., Humic acid and Fulvic acid will activates the aquaculture
pond ecosystem through physical, chemical and biological reactions at general pond pH values.
It strengthen s the metabolic processes in all living things i.e. plankton, live feeds of
aquaculture organisms and aquaculture organisms.
The one of the important property of Pond is the ability to flocculate the organic ions by
ionic, complex, chelate and polar adsorption. This will make the pond waters free from turbidity,
hence keeps the stable growth of plankton growth and enhances the free diffusion of oxygen from
atmosphere into the pond water .It also strengthen the immune system and increase the animal
resistance against most important diseases as well as increasing the live weight of the culture
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animal. Humic substances by their phagocytosis activity inhibits the pathogenic bacterial growth and
growth of moulds, thus decreasing levels of disease incidences and mycotoxins. The humic contents
having the ability to neutralize the negative effects of pesticide residuals and toxins in the ponds.
The probiotics organism of Pond can stable the plankton blooms in aquaculture ponds by
enhancing the availability of nitrogen and phosphorous. Nitrogen is vital for plankton growth. It is
abundant in the soil organic matter (SOM), but not in a form that plankton can use. Nitrogen fixing
bacteria convert SOM's nitrogen to an inorganic form that is useable by plants. In addition, they
provide residual nitrogen, in the soil, which can reduce or eliminate nitrogen fertilizer requirements.
Nitrogen-fixing bacteria of Pond are also capable of combining free nitrogen of the water or air with
oxygen by other species of bacteria that live freely in the soil (no symbiotic nitrogen-fixation). The major
conversion of N2 into ammonia and hence into proteins, is achieved by microorganisms in the process
called nitrogen fixation (or denitrogen fixation). Phosphorous-Solubilising bacteria solubalized
phosphorous for plankton and friendly bacteria from insoluble mineral sources. Pond formulated,
through Natural Systems Approach, to restore a productive system in distressed and depleted aquaculture
ponds.
Experiment No.14
Date:
BIOMETRY OF FISH
Aim: To study the meristamatic characters of fish
Introduction:
Morphometric and Meristic Characters
Measuring the linear dimensions of the whole or part of a fish is probably the most widely used
technique in fisheries biology studies. Morphometric measurements are• any standard measurements that can
be taken on a fish such as Standard Length, Snout Length, length• of largest fin ray of the dorsal fin, depth
of the caudal peduncle and so on. Since these measurements change as the fish grows, these are usually
expressed as ratios to Standard Length. Such ratios are only useful if comparisons are made between
samples of fish of approximately the same size and sex, since the growth of a fish is not always proportional
in all directions and sexual dimorphism is also noticed among fishes. Thus Morphometric measurements
while vital for describing fish species may be of limited usefulness.
Material,
1. Foot scale
2. Thread
3. Divider
4. Dissection microscope
5. Fish
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Method:
With the help of foot scale various measurements were taken as shown in the results.
Body Measurements or the identification of fish's body measurement and fin formula are most important.
With the help of the above the fish can be identified up to species level easily. The following are the different
measurements used for fish identification.
Fish measurements
1. Total length (A-J): It is measured from tip of the snout to end of caudal fin.
2. Standard length (A-H): It is measured from tip of the snout to base of the caudal fin.
3. Head length (A-D): It is measured from tip of the mouth to end of operculum.
4. Snout length (A B): It is measured from tip of the snout to the anterior margin of the eye.
5. Predorsal length (A-E): It is the distance between tips of the snout to origin of the dorsal fm.
6. Preceptor length (A-D): It is the distance between tips of the snout to origin of pectoral fin.
7. Pre pelvic length: It is the distance between tips of the snout to origin of pelvic fin.
8. Pre anal length (A-G): It is the distance between tips of the snout to origin of anal fin.
9. Length of caudal peduncle (G H): It is the distance measured from the posterior base of the anal fin up to
origin of caudal fin.
l3 c ll EF
10. Height of the caudal peduncle: It is measured vertically through the body at caudal peduncles
narrowest part.
11. Height of the body: It is measured vertically through the body at its deepest part.
12. Diameter of Eye: It is measured from one margin of the orbit to other.
13. Inter orbital length: It is distance between two orbits on dorsal surface.
14. Fin measurements: The length to pectoral fm, pelvic and caudal fin is measured long their longest
fin ray.
15. Profile of the body: It gives the outline of the body of fish, along its dorsal and ventral surfaces.
16. Barbles: The number of barbules ranges from 1-4 pairs. These are named according to their position
as nasal, rostral, maxillary and mandibular.
17. Branchiostegal rays: These are slender bony rods found on inner surface of operculum.
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18. Lateral line: This line is found on both the lateral sides of the body. It is a longitudinal row of
perforations of sense organs. It may be complete, incomplete or interrupted.
19. Scales: The scales are supposed to be identifying card of fish. These are also useful in identification of fish
up to species level. The scales are counted along the Lateral line and their number is written after the
abbreviated as L1 .In case the lateral line is absent, the scales are counted along throw where the lateral lines
might has been located and their number is written after the abbreviation as L.r. the transverse rows of scales are
counted from the anterior base of dorsal fin to the ventral line and their number is mentioned after the
abbreviation as L.tr. In this case the scales above and below the lateral line are separated by an oblique (I)
stroke. Pre dorsal scales are counted from anterior extremity to the origin of dorsal fm.
20. Fin formula: Fin formula is constructed after counting the fin rays of pectoral (P), Pelvic (V), Dorsal
(D), ANAL (A), and caudal(C) fins and Lateral line scales. This formula provides scientific information to
con firm the actual identity of a particular fish. The number after the abbreviation of fm denotes number of
fin rays. An oblique (I) stroke indicates the spiny and soft rays of fin.
Results
S.No. Parameter Value
1 Body weight gr
2 Body volume ml
3 Type of scales
4 Total length. It is measured from tip of the snout to end of caudal fin
5 Standard length: It is measured from tip of the snout to base of the caudal fin.
6 Head length: It is measured from tip of the mouth to end of operculum.
7 Snout length: It is measured from tip of the snout to the anterior margin of the eye.
8 Predorsal length : It is the distance between tips of the snout to origin of the dorsal
fin.
9 Preceptor length It is the distance between tips of the snout to origin of pectoral
fin.
10 Pre pelvic length It is the distance between tips of the snout to origin of pelvic fin.
11 Pre anal length : It is the distance between tips of the snout to origin of anal fin.
12 Length of caudal peduncle: It is the distance measured from the posterior base of
the anal fm up to origin of caudal fin.
13 Height of the caudal peduncle : It is measured vertically through the body at
caudal peduncles narrowest part.
14 Height of the body : It is measured vertically through the body at its deepest part.
15 Diameter of Eye It is measured from one margin of the orbit to other.
16 Inter orbital length: It is distance between two orbits on dorsal surface.
17 Fin measurements: The length to pectoral fm, pelvic and caudal fin is measured
long their longest fm ray.
18 Types of fms
19 Barbles: The number of barbules.
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51
20 Branchiostegal rays: These are slender bony rods found on inner surface of
operculum.
Fin Formula: D. 16; P l. 17; P2. 9; A. 8

Discussion;
From the data we can conclude that the fish we have studied is the mrigal carp (Cirrhinus cirrhosa,
also Cirrhinus mrigala, also known as the mrigal and the white carp, is a species of ray-finned fish in the
carp family. Native to streams and rivers in India, the only surviving wild population is in the Cauvery River,
leading to its IUCN rating as vulnerable. It is widely aqua farmed and introduced populations exist outside its
native range It reaches a maximum length of 1 m (3.3 ft).
Mrigal is popular as a food fish and an important aquaculture freshwater species throughout South
Asia. It is widely farmed as a component of a polyculture system of three Indian major carps, along with
Rohu Labeo and the catla. The introduction to aquaculture across India started in the early 1940s and in the
1950s and in the 1960s to other Asian countries. The mrigal carp fails to breed naturally in ponds, thus
induced breeding is done.
The Indian carps are considered as a delicacy compared to other exotic carp species also cultured in
Asia, and sell for higher prices
Mrigal is the bentho-pelagic and potamodromous plankton feeder. It inhabits fast flowing streams
and rivers, but can tolerate high levels of salinity. Spawning occurs in marginal areas of the water bodies
with a depth of 50 to 100 centimeters (20 to 39 in) over a sand or clay substrate. A 6-kilogram (13 lb)
female can lay a million eggs. This fish has a rapid growth rate; by the age of two individuals can reach a
length of 60 centimeters (24 in) and can weigh as much as 2 kilograms (4.4 lb).
Experiment No.15.
Date:
BIOMETRY OF PRAWN
Aim: To study the morphology of prawn/shrimp
Introduction
Penaeid shrimps are important resources for worldwide fisheries and aquaculture. Morphological
variation studies within and among species penaeid shrimps in nature began over 35 years ago for purposes of
fishery management. The results having been taken from these studies can be used for the purpose of
aquaculture. The study of the morphological characteristics of shrimp species and the analysis of
morphological relationships between broodstocks can be also a useful tool for their management because
identification and discrimination of broodstocks are essential to successful rearing programs.
Penaeid shrimps differ in a variety of morphological characteristics that are the expression of genetic
differences among them. There are, of course numerous studies of the morphological differences among
species which can be used for taxonomic distinctions. These studies generally focus on the structures of
genitalia, appendages, rostra and sculpturing of the carapace. Studies of the variation among the species
morphological characters related to size, shape or other commercial characters are quite limited. Dimensions
collected from photographs of shrimp in either dorsal or lateral view either dead or alive have been used in
studies. Morphometric characters have been successfully used for taxonomic inferences.
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PALAEMON
1. Length cm
2. Wight gr
3. Volume ml
4. Number body divisions = 2
5. Name of the body divisions = 1. Cephalothrax
2. Abdomen
6. Segments in cephalothorax = 13
7. No of segments in cephalic region of cephalothorax =5
8. No of segments in thorax region of cephalothorax =8
9. No of segments in abdomen region =6
x swimmerets
pore ippads (pfeopods)
10. Location of mouth = The slit-like mouth opens mid ventrally at the anterior end of
the
Cephalothorax.
11. Location of anus = Anus is a longitudinal aperture present at the base of the Telson,
Ventrally.
12. Location of female pores = paired female genital apertures open on the inner
surface of
coax of the third pair of walking legs in the female.
13. Location of male genital pores = The paired male genital apertures are present on
the
Inner surface of coax of the fifth pair of walking legs in the
male.
14. Location of the stat cysts= they are on the basal segment of each antennules.
Dr.C.V.Narasimha murthy .aquaculture practice manual 2016-2017 ; M.Sc. Zoology Final year 3rd
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53
Cephalic appendages :
15. Antennules: The protopodite is 3 segmented, with basal structure precoxa.coxa and
basis. In the precoxa a sta t o cys t is present. On the basis 2 long, many segmented whip
like feelers are present. They are tactile sense organs. They are not homologous to expedite
and endopodite. The outer feeler is further divided into an inner smaller branch and outer
larger branch.
B
A) 4TT N .'LkL
ANTENNA
C) MAND11 L5
D) I MAXILLA 1 } P R E coxft ) co , 3 } A . 5 5 4 ! E : w - F c I T E
5} EN C-PO t TE & ) i a r.SDM P RO ES .S • l { o L . V Z D E S S
E) 11 MAXILLA
16. Antenna:
a) The protopodite shows coax and basis.
b) Endopodite is long feeler like structure, which is a tactile sense organ.
c) The exopodite is plate like and it is called Squama.
d) It works as a balancer during swimming.
e) At the base of the coax renal opening is present.
f) Antenna is sensory, excretory and balancing in function.
17. Mandibles:
a) They are present on either side of the mouth.
b) The basal part of coax is divided into two parts, it shows a mandibular and
incisor process.
c) The mandibular process shows 5 or 6 dental plates.
d) The incisor process shows 3 teeth. On the outer margin of the head a mandibular
palp is present, which represents the basis and endopodite.
e) The exopodite is absent. The mandibles are masticatory in function.
18. I Maxilla or Maxilla :
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54
a) The protopodite is 2 lobed. They are called Gnathobases.

b) The endopodite is slender. Exopodite is absent.
19. II Maxillae:
a) The protopodite is fait and is divided into 4 lobes.
b) Endopodite is small and unsegmented.
c) The exopodite is broad, and plate like structure.
d) It is called Scaphognathite or baler. It is useful to bring in water into the
bronchial region.
e) It is helpful for respiration and manipulation of food.
Thoracic appendages :
a) They are 8 pairs.
b) The first 3 pairs are Maxillipeds.
c) The remaining 5 are walking legs.
20. I Maxillipeds:
a) They are thin and leaf like.
b) Protopodite is 2 segmented. The endopodite is short.
c) Exopodite is present.
d) It is bibbed. Epipodite is respiratory in function.
e) It is present on the outer side of coax.
A ) m A x I ._ L I + e D E 1 1 ,
1A4rLLIP55
) C0Str1 2 ) t,SIS E?40•PO3ITe 4 ; hDOPODITE
C) III Mrl7CIUr eDe
) CHFL T E LECS
5 ) E ? I O D P T E fir) L~ILL 1) r S C H I L / .
) .w r , E l - ~ l T
]r~. ups ) erg its ~c) P R C ? G 2 ' J . . s L} AsCrrLlls
21. II Maxillipeds:
a) It has 2 segmented protopodite.
b) Coxa bears a conical epipodite and a gill Endopodite is 5 segmented.
c) The five segments are isocheim, merus, carpus, propodus and dactylus. Exopodite
is long and unsegmented.
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22. III Maxillipeds :

a) It looks like a walking leg. It has 3 segmented endopodite.
b) The basal segment corresponds to isocheim and merus.
c) The apical segment is fused and corresponds to propodus and dactylus.
d) The middle one is carpus.
23. Walking legs:
a) They are 5 pairs.
b) The first 2 pairs are chelate and the other 3 pairs are non chelate.
c) They are useful for walking.
d) The typical walking leg has a two jointed protopodite and 5 jointed endopodite.
e) The protopodite has two segments, coax and basis.
f) The endopodite has isocheim, merus, carpus, propodus and dactylus.
g) In the first and second pairs of legs the propodus is prolonged beyond its articulation
with dactylus and it looks like a chelae or pincer.
h) Such legs are called chelate legs. They catch the food and push it into the mouth.
i) The second chelate, leg in male is larger and powerful than in females.
j) The 3rd. 4th and 5th walking legs are non chelate.
k) In female the 3rd walking leg bears a female reproductive opening on the inner side of
coax
1) In the male the genital opening is present on the arthroidal membrane between the
thorax and 5th walking leg.
24. Abdominal Appendages:
1) Abdomen bears six pairs of appendages.
2) Each appendage is biramous. These are called pleopods or swimmerts.
3) The protopodite has coax and basis. The basis bears two flat leaf like exo and endopodite.
4) From the inner margin of the endopodite a small appendix intema arises
C cO,
.4SaS
G XOPObITU
OPOITU
A PE
T1 r A
1tiP6 N I X
m A S O L.IHA
1) In the females during breeding season the appendix intema of opposite appendages unite
and carry eggs.
2) In the first pair of abdominal appendages the appendix intema is absent.
3) The second abdominal appendages of male shows appendix masculine also.
semester
4) The sixth pair of abdominal appendages will be called Uropods or tail feet.
5) They are large and lay one on either side of the Telson.
6) The two uropods and Telson together form a broad tailfin.
7) It helps the Prawn to take a backward spring in water.
8) In a uropod the coax and basis fuse together to form a triangular sympod. It helps the
Prawn to take a backward spring in water.
9) In a uropod the coax and basis fuse together to form a triangular sympod. It bears exo and
endopodites.
Discussion:
Morphometric studies of the prawn are useful in identification, classification and rearing.
Experiment No.16
Date:
I. STUDY OF GENERAL CHARACTERS AND IDENTIFICATION OF

IMPORTANT SHRIMP/PRAWN.
PENAEUS MONODON
semester
55 Antenna
DorsomTdian ,4Ddoman ! g-Hants (1), tap half c a M

"
a!ina 4 o ) "Lir. u n , not m hair caliatE "pIE'uron
orStjlate€ai
SmIcus ( avlty)
Penaeus monodon:
1. This called as tiger prawn.
2. On ventral side of the body white grayish colour cross strips are present.
3. On ventral side of rostrum 1 to 3 teeth are present.
4. On dorsal side of the rostrum 7-8 dorsal teeth are present.
5. On cephalo thorax hepatic and antennary spines are present.
PENAEUS INDICUS:
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ADROSTRAL CARINA
-FPIGASTRIC SPINE
GASTR0-0RBITAL CARINA -T-IE?ATTC SPINE
HEPATIC CARINA
AHFCENNAT. FLAGELLUM
1. This is called white prawn.

2. It is white in colour.
3. Second ventral plural segment is extended upto first segment.
4. Dendrobranch type of gills is present.
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MACHROBRACHIUM ROSENBERGI
Macrobrachium rosenbergil:
1. This is called as scampi
2. Rostrum is sward shaped with 13 to 14 dorsal spines.
3. Second chelate leg is lengthy in males.
4. Sexual dimorphism is present.
5. Second ventral pleura are extended over to third segment.
6. Phyllobrach types of gills are present.
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Macrobrachium Malcomsoni
1. This is a fresh water prawn.

2. Second pair of legs in males are lengthier than body
3. second ventral pleura are extended over to third segment.
4. Phyllobranch type of gills are present
5. Second waling leg is having complete carapace
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1. Penaeus vannamei grows to a maximum length of 230 millimeters (9.1 in),

2. Carapaces length of 90 mm
3. The rostrum is moderately long, with 7-10 teeth on the dorsal side and 2-4 teeth on
the ventral side.
4. External structures of the genital system are the major dimorphic features. The male
has two pairs of modified abdominal appendages on the first and second abdominal
segments (the petasma and appendix masculina) that deliver sperm to the female's
external receptacle (the thelcum) located between the bases of the fifth walking legs.
5. The petasma, appendix masculine and thelcum are located on the ventral surface
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Experiment No.17
Date:
STUDY OF GENERAL CHARACTERS AND IDENTIFICATION OF

IMPORTANT CARPS
MAJOR CARPS
1. LABO ROHITA
2. CATLA CATLA
3. CIRRHINUS MIRGALA
MINOR CARPS
4. LABEO CALABASU
5. LABEO BATA
6. LABEO FIMBRINATUS
7. PUNTIS CARNATICAS
MURREL FISHES
8. CHANNA PUNCTATUS
9. CHANNA STRITUS
10. CHANNA MARULIUS
CAT FISHES
11. CLARIUS MACROCEPHALUS
12. HETEROPNEUSTES FOSSILIS
13. ANABAS TESTUDINIS
14. ETROPAS SURATENENSES
15. WALLAGO ATTU
16. MYSTUS OAR
17. MYSTUS SENGHALA
18. PANGASIUS UPIENSIS
19. SILONIA SILONIA
EXOTIC FISHES
20. HYPOPHTHALMYCHTHYS MOLITRICUS
21. CTENOPHARYNGODON DELLA
22. CYPRINUS CARPIO
23. OSPHRONEMUS GORAMY
24. TILAPIA MOSSAMBICUS
COLD WATER FISHES
25. SALMO TRUTTA FARIO
26. SALMO GAIRDINERI
27. TOR TOR
28. TOR PUTITORA
29. TOR KHUDRRI
30. TINCA TINCA
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1. LABEO ROHITA
Phylum : Chordata
Class : Osteichthyes
Subclass : Actinopterygii
Order :Ostreophysis
Family : Cypriniformes
Genus : Labeo
Species : :rohita
1. It grows upto 90 cm.

2. Head is clear and ends with stout
3. Ventral side is round
4. Dorsal side is having light dark colour
5. Thin lips are [present
6. Dorsal, ventral caudal & anal fins are present
D16(3/13);P17; V9; A7(2/5);C19;LI4o41;Ltr6 Y2- 79/2-8 B.1pairs
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2.
CATLA CATLA
Phylum : Chordata
Class : Osteichthyes
Order :Ostreophysis
Genus : Catla
Species : Catla
1. Dorsal fin is bigger and anal fin is small

2. No upper lip. Lower lip is stout
3. Spiracle is present on the dorsal side
4. Caudal fin is having spine
D 3/15 16 P
1819( - ); 9 i V 9 ; A8(3/5);C19;L143;Ltr7 %4 61/7 - B.1pairs
66
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3. CIRRAHNAS MRIGALA
Phylum : Chordata
Class :Osteichthyes
Order :Ostreophysis
Genus : Cirrhinus
Species : mrigala
1. Small head with soft body is present

2. At the anterior end small round shaped mouth is present.
3. Dorsal fin is made up of 12-15 fin rays.
4. Fins are orange colored.
5. Maxillary barbules are clear.
6. Dorsal side is black and rest of the body is grayish colour.
D16(3/13;P18i V9; A8(2/6);C15iL1X;Ltr6 %-/ 61/2- B.1pairs
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4.. LABEO CALBASU
Phylum : Chordata
Class :Osteichthyes
Order :Ostreophysis
Genus : Labeo
Species : :calbasu
1. The body is bluish green in colour with small head and folded lips.
2. Head is long conical with a stout
3. The snout consists of four black colored long barbs.
4. It is Culturable in pond
5. . It reaches to a size of 1 m and 1 .5 to 2 kg. in weight
Fin formula
D17(3/14);P19; V 9 ; A7(2/5);C19;LI41iLtr7 %2-8 B.2pairs.

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5. LABEO BATA:
Phylum : Chordata
Class :Osteichthyes
Order :Ostreophysis
Genus : Labeo
Species : :bata
1. The body is elongate and 160 cm.

2. Its dorsal profile is more convex than the ventral.
3. The snout slightly projects beyond the mouth, often studded with pores (Talwar and
Jhingran, 1991).
4. A pair of small maxillary barbells is hidden inside the labial fold.
5. There is no cartilaginous support to the lips.
6. The dorsal originates midway between the snout tip and the anterior base of anal.
7. Pelvic originate slightly nearer to the snout tip than to the caudal base (Rahman,
1989).
8. It is bluish or darkish on upper half, silvery below, and the operculum is light
orange.
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6. LABEO FIMBRIATUS:
Phylum : Chordata
Class :Osteichthyes
Order :Ostreophysis
Genus : Labeo
Species ::Fimbriatus
1. It has folded lips and lives in deep water zone.

2. It grows to a maximum size of 90 cm and 450 g. in weight.
3. Red spots are present on the scales of middle row.
4. Dorsal fin is made up of 12-15 fin rays.
5. Fins are orange colored.
6. Maxillary barbules are clear.
7. Dorsal side is black and rest of the body is grayish colour.
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7 Puntius carnaticus
Phylum: Chordata Class:

Actinopterygii Order:
Cypriniformes
Family: Cyprinidae
Genus: Hypselobarbus Species: H. carnaticus
1. Puntius is a genus of cyprinid fishes known as the spotted barbs for the
predominant pattern, though many have vertical black bands instead.
2. The maximum size for an adult of this genus is less than 25 cm (9.8 in),
typically 7-15 cm (2.8-5.9 in), and achieve around 5 cm (2.0 in) adult length.
3. In appearance they may resemble miniature carp and are often brightly colored
or patterned.
4. It is also called as Karnataka carp
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8 CHENNA PUNCTATA
Phylum : Chordata
Class :Osteichthyes
Order :Ophiocephali
Family
Genus : Chenna
Species : punctata
After Meeker, 1878
1. Head is snakehead shaped.

2. Body is lengthy and semicircular
3. Spines are not present.
4. Single dorsal fin is present.
5. Anal fin is smaller than dorsal fin
6. Caudal fin is round shaped.
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9 CHENNA STRAITUS
Phylum : Chordata
Class :Osteichthyes
Order :Ophiocephali
Family
Genus : Chenna
Species : striates
dorsal fin trunk
pelvic fin scales
Fig. 154. Channa striates.

2. Dark striations are present on the body
3. Body is lengthy and semicircular
6. Anal fm is smaller than dorsal fin
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10 CHENNA MARULIAS
BULLSEYE SNAKEHEAD OR GREAT SNAKEHEAD

Phylum Chordata
Class :Osteichthyes
Order Ophiocephali
Family
Genus : Chenna marulias
Species

2. Six dark large spots are present on the body
3. A spot is present on the caudal fin
6. Anal fin is smaller than dorsal fin
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11. CLARIES MACROCEPHALUS
Phylum : Chordata
Class :Osteichthyes
Order :Ostreophysis
Family : claridae
Genus : claries
Species :cephalus
1. This is a cat fish.

2. Body is dorso ventrally compressed.
3. 4 pairs of barbules are present.
4. Dorsal fin is lengthy and spine less.
5. No dentition in mouth.
6. No scales are present on the body.
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12. HETEROPNEUSTES FOSSILIS

CAT FISH
Phylum: Chordata
Class: Actinopterygii
Order: Siluriformes
Family: Heteropneustidae
Genus: Heteropneustes
Species: H. fossilis
1. Body is divisible into head, trunk, and tail

2. Mouth is transverse
3. Four pairs of barbules are present
4. Fins are dorsal, pectoral, ventral, anal and caudal.
5. Teeth are present jaws
6. Long anal fin is present
Fin formula : D6;P 1/7;v:6; A62-66;c;19
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13 . ANABAS TESTUDINEUS
Phylum : Chordata
Class :Osteichthyes
Order : percophormi
Family : anabantidae
Genus : Anabas
Species : testudineus
Climbing perch, Ana has testudineus
1. Mouth is terminal
2. Body shape is fusiform or normal
3. No of scales on the later line are 36.
4. Dorsal fin rays are 16-20
5. Caudal fin is truncate
6. Scales large and regularly arranged, ciliate.
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14 . ETROPLUS SURATENSIS
PERAL SPOT

Perciformes Family:
Cichlidae Genus: Etroplus
Species: E. suratensis
1. The adult is oval in shape

2. Snout is short
3. Fish is gray-green in color with dark barring and a dark spot at the base of the
pectoral fin.
4. It commonly reaches 20 centimeters (7.9 in) in length, and the maximum length is
twice that.
5. This species lives in brackish water habitat types, such as river deltas. It eats mainly
aquatic plants,
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15. WLLAGO ATTU
Phylum : Chordata
Class : Teleostei
Subclass : Cypriniformes
Order :Ostreophysis
Family ailuridae
Genus : Wallgo
Species : attu
1. This is called as fresh water shark.

2. Body is dorsally compressed.
3. At the anterior [part of the body bog mouth is present up to eyes.
4. Big head small body and lengthy tail is present.
5. Two pairs of barbules are present.
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16. MYSTUS OAR
Chordata
Phylum: Actinopterygii
Class: Perciformes
Order: Cichlidae
Family: Etroplus
Genus:
Species: E. suratensis
longer
upper jaw
F / maxillary
elongated
pectoral fin maxillary barbel
pelvic fin
1. Fish measures about 17 -27 cm

2. Snout is broad and spatulate
3. Upper jaw is longer than the lower jaw
4. Four pairs of maxillary barbules are present
5. Caudal fin is deeply forked
6. A large round black spot is present on the inferior part of dorsal fin
Fin formula: D.1/7/10. V.6;A.13(3/10);C17
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17 . MYSTUS SEENGHALA
Kingdom: Animalia
Phylum: Chordata
Order: Siluriformes
Family: Bagridae
Genus: Mystus
F i g . 145. A11 s t c ls s e e u g l i a l a .
1. Fish measures about 15 -30 cm

2. Snout is spatulate
3. Teeth on the palate in the form of continuous crescent
4. Upper jaw is longer than the lower jaw
5. Four pairs of maxillary barbules are present
6. Caudal fin is deeply forked
7. A large round black spot is present on the inferior part of pelvic fin
Fin formula: D.1/7/10. P. 1/9;V.6;A.11-12(3/8-9);C19-21.
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18 PANGASIUS UPENSIS

Siluriformes Family:
Pangasiidae Genus:
Pang asius
1. Fish measures about 90-95 cm

2. Upper jaw longer than lower jaw
3. Broad anal fm is present
4. Two pairs of barbules are present
5. Mandabularis is as long as head.
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semester
19. SAILONIA SILONIDA
Kingdom: Animalia
Phylum: Chordata
Order: Siluriformes
Family: Schilbeidae
Genus:
Silonia
Species:
S. silondia
1. Body elongated and deeply compressed.

2. Mouth terminal and lower jaw little longer.
3. Snout broad and rounded.
4. Eyes with narrow adipose lids. Barbels 2 pairs. Dorsal spine comparatively weak
than pectoral spine and both spines finely serrated posterior.
5. The body color is yellowish-green on back, silvery purple on flanks and abdomen,
golden tinge present on both sides of head. Caudal, anal and pelvic bases yellowish.
Fin formula:
D1. I/7; D. 0; P. I/11-13; V. 6; A. 40-46 (4/35-44); C. 17
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20 . Hypophthalmicthys molitirx
Silver carp:
Phylum- Chordata
Class-Osteichthyes
Order- Cypriniformes
Family- Cyprinidae
Genus-
Hypophthalmichthys
1. Body is stout and compressed, with a sharp keel from throat to vent.
2. Head rather small, post operculum with radiated strides; snout blunt, obtusely
rounded anteriorly.
3. Mouth is terminal, lower jaw slightly longer than upper.
4. Dorsal fin is short, in serrated slightly behind pelvic fins, scales small.
5. Colour silvery-white with blood red spots on body especially on caudal peduncle.
Fins are dark.
Fin formula:
D. iii 7; A. i-iii 12-14 P i 17; V i 7 (Talwar and Jhingran, 1991) D 3/7; P11/17; P21/7; A 2-3/12-14
(Rahman and Ruma, 2007)
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21. CTENOPHARYNGODON IDELLA Phylum: Chordata
Order: Cypriniformes
Family: Cyprinidae
Subfamily: Leuciscinae
Genus: Ctenopharyngodon
Species: C. idella
1. Grass carp have elongated, chubby, torpedo-shaped body forms.

2. The terminal mouth is slightly oblique with non-fleshy, firm lips, and no barbels.
3. The complete lateral line contains 40 to 42 scales.
4. Broad, ridged, pharyngeal teeth are arranged in a 2, 44, 2 formula. The dorsal fin
has eight to 10 soft rays, and the anal fin is set closer to the tail than most cyprinids.
5. Body color is dark olive, shading to brownish-yellow on the sides, with a white
belly and large, slightly outlined scales.
6. The grass carp grows very rapidly. Young fish stocked in the spring at 20 cm (7.9
in) will reach over 45 cm (18 in) by fall. The average length is about 60-100 cm
(23.5-39.5 in). The maximum length is 1.4 m (4.6 ft) and they grow 40 kg (881b).
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22.CYPRINUS CARPOI
Kingdom: Animalia
Phylum: Chordata
Family: Cyprinidae
Genus: Cyprinus
Species: C. carpio
1. 2 pairs of barbells; dorsal fm with 15-20'A branched rays;

2. caudal fm deeply emarginated
3. Pharyngeal teeth 1, 1, 3:3, 1,1, robust, molar-like with crown flattened or somewhat
furrowed. Scales large and thick. 'Wild carp ' is generally distinguished by it's less stocky
build with height of body 1:3.2-4.8 in standard length.
4. Very variable in form, proportions, squamation, development of fms, and color.
5. Caudal fin with 3 spines and 17-19 rays (Ref. 2196). Last simple anal ray bony and
serrated posterior; 4 Barbels; 17-20 branched dorsal rays; body grey to bronze
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23.OSPHRONEMUS GORAMY
Phylum: Chordata
Order: Perciformes
Family: Osphronemidae
Genus: Osphronemus
Species: O. Goram y
1. With 8-10 complete dark vertical bars in juvenile color phase;

2. Adults without vertical bars or sexual dichromatic,
3. Both sexes drab;
4. Transverse scale rows usually 6.1.12; dorsal fin spines usually 12-13 (rarely 11 or
14); soft-rayed portion of anal fin greatly enlarged, its distal margin parallel to
distal margin of caudal fin;
5. Caudal fin rounded or obtusely rounded, not truncate or emarginated
6. Pelvic fins with first soft ray prolonged into a thread-like tentacle reaching
posterior to or beyond hind margin of caudal fin.
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24. Tilapia mossambica
Sarotherodon mossambicus
Phylum: Chordata
Order: Perciformes
Family: Cichlidae
Subfamily: Pseudocrenilabrinae
Tribe: Tilapia
Genus:
Oreochromis
0
41,1
I I I F+ 1 i ii1 4 4 4 s I l•
1. Body compressed; caudal peduncle longer than deep. Scales cycloid.

2. A knob-like protuberance present behind upper jaw on dorsal surface of snout.
3. Upper jaw length shows sexual dimorphism, and mouth of male larger than that of
female.
4. First gill arch with 20 to 22 gillrakers. Lateral line interrupted. Spinous and soft
ray parts of dorsal fin continuous.
5. Dorsal fin with 15 to 18 spines and 10 to 13 soft rays.
6. Anal fin with 3 spines and 9-10 rays. Caudal fin truncated.
7. Colour in spawning season, pectoral, dorsal and caudal fins becoming reddish;
colour male shows much brighter orange tail than female.
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25. SALMO TRUTTA
Phylum: Chordata
Class:
Actinopterygii
Order:
Salmoniformes
Family: Salmonidae
Genus: Salmo
Species: S. trutta
1. Trout have fins entirely without spines, and all of them have a small adipose fin
along the back, near the tail.
2. The pelvic fins sit well back on the body, on each side of the anus.
3. The swim bladder is connected to the esophagus, allowing for gulping or rapid
expulsion of air, a condition known as physostome.
4. Unlike many other physostome fish, the trout do not use their bladder as an
auxiliary device for oxygen uptake, relying solely on their gills.
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26.
SALMO GIRDINERI
Phylum: Chordata
Order: Salmoniformes
Family: Salmonidae
Genus: Oncorhynchus
Species: O. mykiss
f '!s'!1 tt IF 4 q '" r r :
,w ‘ V
y , a t t * aY.~ Hr is !'w . , w a
hili $ Aa 44iK«...:~=
. t o 0A k to* 4 * - .4
*
+n
'
a M rl r' 7 t' 9 *4.0. r' rfI~ Ys
A i i, N
1 1fIA 1o w '~ + »
te ,~' M~ M a r a 0• ,. *a.
•,v cA ,O
1. The rainbow trout can be distinguished by its pattern of dark spots on a light
background. See also similar species information.
2. Mouth and snout: Terminal, large and slightly oblique, with numerous small to
medium-sized teeth on the upper and lower jaws. No Barbels. Body patterning,
color, and scales: Spots on side, with an obvious broad red to pink lateral stripe in
inland populations; juveniles also with oval dark vertical bars ("parr marks").
3. Inland populations are densely spotted with a background of dark olive or green on
the back, silver, bronze or olive on the sides, and cream below, with a broad, red,
rose, pink, or purple lateral stripe.
4. Dorsal and tail fins tan, olive, or gray; both usually spotted throughout. Pectoral,
pelvic, and anal fins tan, olive, or gray, usually without spots. Adipose fin spotted
but not edged in red or orange.
5. Body shape and size: Body fusiform; oval in cross section. Inland typically 300-400
mm (12-16 in) TL, maximum about 550 mm (22 in). Great Lakes typically 500-700
mm (20-28 in) TL; maximum about 800 mm (32 in).
6. Tail, dorsal and other fins: Slightly forked to square tail. Single dorsal fin with no
spines and 10-12 principal rays. Pelvic fins abdominal with axillary process.
94
A
dipose fin present. Anal fin with 8-12 principal rays.
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27 . TOR TOR

Cypriniformes Family:
Cyprinidae Genus: Tor
Species: T. tor
1. The body is stout, elongated and compressed.

2. The ventral profile is more arched than dorsal profile. Head is comparatively
smaller and pointed. Mouth small and inferior.
3. Eyes visible from below the head. Lips are thick and fleshy, running at angles of
mouth. Barbels two pairs and scales large.
Lateral line scales 25-26 (Rahman, 1989) and 22-27 (Talwar and Jhingran, 1991).
4. A scaly sheath is present on the base of dorsal and the last unbranched ray of dorsal
fin is comparatively strong, smooth and osseous.
5. Caudal fin is deeply forked. Color of the dorsal side is dark grey.
6. Golden to pinkish on flanks and abdomen is silvery with slightly golden tinge.
7. Below the eyes is light yellow. Dorsal, pectoral and anal fins are reddish yellow in
color.
Fin formula
D. 12 (3/9); Pi. 16-17; P2. 9; A. 7 (2/5) (Rahman, 1989)
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28. TOR PUTITORA

Species: T. putitora
1. the body is elongated and both profiles (dorsal and ventral) nearly straight and
somewhat compressed.
2. Mouth small and upper jaw slightly longer than that of lower jaw.
3. Lips thick and fleshy. Barbels two pairs.
4. Last unbanked ray of dorsal fin is comparatively strong, smooth and osseous.
5. Pelvic fins contain a scaly appendage. Caudal deeply forked. 25-28 (Talwar and
Jhingran, 308); 25-26 (Rahman, 1989) scales on lateral line.
6. Color of the side is greenish silvery. Belly silvery to white.
7. Scale golden with dark base and formed of minute black spot. Anal, pelvic and
pectoral fins reddish yellow in color.
Fin formula:
D. 2/9; P1. 15; P2. 9; A. 2/5 (Rahman, 1989 and 2005)
Maximum lengths and weights: 23-26 cm (Rahman, 1989), 270 cm (Talwar and
Jhingran, 209). 54 kg fish was recorded from Cauvery river; 24.4 kg from Teesta and
Riyang rivers in northern Bengal (Rahman, 1989).
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29. TOR KHUDREE

Species: T. khudree
1. Colour silvery or greenish along the upper half of the body, becoming silvery shot
with gold on the sides and beneath. Lower fins reddish yellow.
2. Length of the head is 4 to 5 inches and the widest point of the body is at 4.3 to
5.5 inches from the snout.
3. The eyes are at 6.25 to 7.5 inches behind the snout in moderate sized specimens but
as much as 3.5 inches smaller specimens.
4. The lips are thick, with an uninterrupted fold across the lower jaw,
5. . The maxillary pair of barbels is longer than the rostral, and extending to below the
last third of the eye.
6. Fins the dorsal arises opposite the ventral, and is three fourths as high as the body;
its last undivided ray is smooth, osseous, strong, and of varying length and
thickness.
7. the spine is very much stronger and as long as the head excluding the snout.
8. Pectoral as long as the head excluding the snout ; it reaches the ventral, which is
little shorter. Anal laid flat does not reach the base of the caudal, which is deeply
forked.
9. Lateral line complete, 2 to 2.5 rows of scales between it and the base of the ventral
fin ; 9 rows before the dorsal.
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30. TINCA TINCA

DOCTOR FISH
Phylum: Chordata
Family: Cyprinidae
Genus: Tinca
Species: T. tinca
1. Tench have a stocky, carp-like shape and olive-green skin, darker above and almost
golden below.
2. The caudal fin is square in shape.
3. The other fins are distinctly rounded in shape.
4. The mouth is rather narrow and provided at each corner with a very small barbel.
5. Maximum size is 70 cm, though most specimens are much smaller.
6. A record fish caught in 2001 in England had a weight of 15 lb 3 oz (6.89 kg).
7. The eyes are small and red-orange in colour.
8. Sexual dimorphism is weak, limited to the adult females having a more convex
ventral profile when compared with males.
9. Males may also possess a very thick and flattened outer ray to the ventral fins.
10. Males are generally smaller than females, but can be recognized by having more
curved lower fins and noticeable muscles around the base of the fins generally
absent in female.
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EXPERIMENT NO.18
DATE:
INDUCED BREEDING IN FISH
Aim ; To learn the technique of induced breeding in fish
Introduction
Induced breeding is a technique whereby ripe fish breeders are stimulated by
pituitary hormone or any other synthetic hormone introduction to breed in captive
condition. The stimulation promotes timely release of sperms and eggs.
The technique of induced breeding was first evolved in Argentina after producing
pituitary extract by Houssay 1930 where viviparous fish was injected with the hormone to
make premature birth. In the year of 1934, Brazilians were succeeded in induced breeding by
pituitary extract. This technique was also followed in America and in Russia. In India first
attempt of induced breeding was made by an in 1937 on Cirrhinus mrigala. Later in 1955 Dr.
Hiralal Choudhuri applied this technique in minor carps (Esomus danricus, Pseudeotropius
atherinoides). Ramaswamy and Sunderaraj first induced to breed Clarias batrachus &
Heteropneustes fossilis. The first successful induced breeding on major carps was done by
Dr. Hiralal Choudhuri 1957- Cirrhinus mrigala, C. reba, & Labeo rohita. Parameswaran &
Alikuni successfully bred the exotic Chinese carps -
Hypophthalmichthys molitrix & Ctenopharyngodon idella in 1963.
Technique of Induced breeding:
Preparation of Pituitary Extract

For preparation of gland extract the glands were removed carefully from freshly
killed fish called donor fish. For best result the donor fish was fully ripe and mature.
Common carp is the best donor fish, because it breeds throughout the year and the
individuals are available in all parts of the world. The pituitary glands of such species are
relatively large. The gland was collected prior to spawning. However the gland doesn't show
species specificity and any carp species can be used as donor. However the glands of relative
or closely related species show best result.
Selection of Brooders:
Proper selection of are the key of success in case of induced breeding. The breeders
should be healthy, fully ripe and of medium sized. They should preferably come into the age
group ranging from 2 - 4yrs and have the weight of 1 - 5kgs. Large sized breeders are
avoided for difficulty in handling. For ripe male and female carps, it can be easily identified.
The male shows roughness on pectoral fins when belly pressed milt freely oozes out. The
ripe female shows relatively smooth pectoral fins and operculum. The eggs are released
when the belly is pressed smoothly in female.
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The belly of ripe female is generally soft and round or budged. The vent is swollen,
protruding and pinkish in colour. It is wiser to practice to keep ready adequate stock of
potential brooders. For this a few months before breeding season potential breeders are kept
away under care, and fed on supplementary feed (rice bran and oil cake mixture).
Injection to the breeders:
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The pituitary extract is administered into the body of breeders by means of hypodermic
syringe either infra muscular or infra peritoneal. To ensure a higher percentage of
fertilisation during induced spawning it is necessary that there is synchronisation between
ovulation and milt shading. This difficult to achieve with a set of breeders having one male
and one female. Therefore the common practice is to use a set consisting of one female and
two males. Determination of correct dosage of pituitary extract to be given to the breeders is
very important though a difficult matter. Dosage depends upon the size and state of maturity
of the recipient (breeders) as well as upon the state of maturity of the donor for the glands. It
has been found that the potency of the gland is influenced by the size, the age, the sex, the
state of sexual maturity of the donor fish as also the size of the gland itself. Great difficulty is
encountered because it is not easy matter to ascertain the state of maturity of fish from
external examination. Usually the female is given a preliminary dose of 2-3mg/kg of body
wt. The preliminary dose is not given to the male. After an interval of time about 6 s a
second dose of 5 - 8mg are given per kg of body wt of female. The male was given then the
first dose of injection with female @ 2-3mg/kg of body wt. The dose may be depending
upon the maturity of fish, age, sex and also the environmental
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conditions.
For infra muscular injection the fish is laid on its side while held in hand net and the
needle is inserted either in the caudal peduncle or in the shoulder. For infra peritoneal the
injections are given in the bases of paired pectoral fins. But it is avoided because less expert
hand can puncture heard of the fish.
DISCUSSION
Reproduction in fishes is regulated by external environmental factors that trigger

internal mechanisms. The final event of the reproductive cycle, the release of eggs and
sperm resulting in spawning, can be controlled by either placing the fish in an appropriate
environ-
Meant or by changing the fish's internal regulating factors with injected hormones or other
substances. The internal mechanisms that regulate spawning are similar for most fishes. The
external environmental factors that control reproduction, however, vary considerably among
species. For this reason, more is known about the internal regulatory mechanism of fish
reproduction than the specific environmental requirements for spawning each species.
Environmental factors that have been shown to play a significant role in the reproductive
cycle.
The internal mechanism that regulates the process of reproduction in fish is the
brain-hypothalamus pituitary-gonad chai.Hormone induced spawning techniques influence
this sequential mechanism at several levels, by either promoting or inhibiting the Process.
The primary substances used for hormone-induced spawning have been: (1) pituitary
extracts and (2) purified gonadotropin to stimulate the ovaries and testes;
Experiment No 19
Date:
INDUCED BREEDING IN PRAWN/SHRIMP
Aim: To artificially induce breeding in Prawn by eye stalk ablation technique.
Introduction
Eyestalk ablation is the removal of one (unilateral) or both (bilateral) eyestalks

from a crustacean. It is routinely practiced on female shrimps (or prawns) in almost every
marine shrimp maturation or reproduction facility in the world, both research and
commercial. The aim of ablation under these circumstances is to stimulate the female
shrimp to develop mature ovaries and spawn.
Most captive conditions for shrimp cause inhibitions in females that prevent them
from developing mature ovaries. Even in conditions where a given species will develop
ovaries and spawn in captivity, use of eyestalk ablation increases total egg production and
increases the percentage of females in a given population that will participate in
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reproduction. Once females have been subjected to eyestalk ablation, complete ovarian
development often ensues within as little as 3 to 10 days.
The most commonly accepted theory of why eye ablation reduces this inhibition is that a
gonad inhibitory hormone (GIH) is produced in the neuro secretory complexes in the
eyestalk. This hormone occurs in nature in the non-breeding season and is absent or present
only in low concentrations during the breeding season. The reluctance of most shrimp to
routinely develop mature ovaries in captivity is a function of elevated levels of GIH, and
eyestalk ablation lowers the high haemolymph titer of GIH. The effect of eyestalk ablation is
not on a single hormone such as GIH, but rather affects several physiological processes.
Besides the GIH evidence, another hypothesis suggests that eyestalk ablation also reduces
light intensity and thereby induces ovarian maturation. In the banana prawn
(Fenneropenaeus merguiensis, syn. Penaeus merguiensis), dim light favours ovarian
maturation and spawning. The exact mechanism of eyestalk ablation on the ovarian
maturation is not conclusive. It has been reported that in the tiger prawn (Penaeus monodon),
the eyestalks fully regenerate in less than 6 months.
Principle
Reproduction, in crustaceans has been hypothesized to be controlled by dual
endocrine factors - a gonad-inhibiting hormone (GIH) from the X organ-sinus gland
complex and a gonad-stimulating hormone (GSH) secreted by the brain/ thoracic ganglion of
maturing females, the actions of which are antagonistic to each other (Adiyodi and Adiyodi,
1970). In the majority of malacostracan crustaceans, eyestalk is the pivotal organ for housing
various neuropeptides responsible in regulating maturation. Eyestalk ablation reduces the
titer of GIH in females causing accelerated ovarian growth. Thus, endocrine manipulation
to induce gonadal maturation has so far been synonymous with unilateral eyestalk ablation
and has a far- reaching impact on crustacean aquaculture. Eyestalk ablated shrimps respond
to their operation with a rapid and unstoppable gonadal development thus augmenting total
egg production in a given time.
Material:
1. Scissors,
2. Forceps
3. Blade
4. Tiger prawn (Penaeus monodon)
Methods:
Techniques used for eyestalk ablation include:
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1. Pinched the eyestalk, usually half to two-thirds down the eyestalk. This method
may leave an open wound.
2. Slitting one eye with a razor blade, then crushing the eyestalk, with thumb and
index fingernail, beginning one-half to two-thirds down the eyestalk and moving
distally until the contents of eyes have been removed. This method, sometimes
called nucleation, leaves behind the transparent exoskeleton so that clotting of
haemolymph, and closure of the wound, may occur more rapidly.
3. Cauterizing through the eyestalk with either an electrocautery device or an
instrument such as a red-hot wire or forceps. If performed correctly, this method
closes the wound and allows scar tissue to form more readily. A variation of this
technique is to use scissors or a sharp blade to sever the eyestalk, and then to
cauterize the wound.
4. Ligation by tying off the eyestalk tightly with surgical or other thread. This method
also has the advantage of immediate wound closure.
5. prawns treated with lignocaine (a local anesthetic in mammals) prior to eyestalk
ablation show less rubbing, flicking and sheltering than those not given the
anesthetic.
PROCEDURE
In the present experiment bilateral eye stalk ablation was made to paeneus
monodon by making cut at the base of eyestalk.
DISCUSSION:
There are several direct and indirect effects of eye ablation in female shrimps, including;
1. Increases total egg production by producing more frequent spawning, but not larger
spawns
2. moult cycle duration will be shorter
3. Increases mortality.
4. Deteriorates female condition
5. In some instances, produces lower hatch rate of eggs
6. Leads to changes in ovarian colour
7. Increases energetic demands
8. Leads to eventual loss in egg quality
References
1. Bray, W.A.; Lawrence, A.L. (1992). Reproduction on Penaeus species in captivity.

In: Fast A.W. and Lester L.J. (Eds). Marine Shrimp Culture: Principles and
Practices. Developments In Aquaculture And Fisheries Science 23 (Elsevier, The
Netherlands). pp. 93-170
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105
2. Hoang T, Lee SY, Keenan CP, Marsden GE (2002). "Ovarian maturation of the
banana prawn, Penaeus merguiensis de Man under different light intensities.".
Aquaculture 208: 159-168.
3. Desai, U.M.; Achuthankutty, C.T. (2000). "Complete regeneration of ablated
eyestalk in penaeid prawn, Penaeus monodon." (PDF). Current Science 79 (11):
1602-1603.
4. Diarte-Plata, G., Sainz-Hernandez, J.C., Aguinaga-Cruz, J.A., Fierro-Coronado, J.A.,
Polanco-Torres, A. and Puente-Palazuelos, C. (2012). "Eyestalk ablation procedures
to minimize pain in the freshwater prawn Macrobrachium americanum". Applied
Animal Behaviour Science 140 (3): 172-178.
EXPERIMENT NO.20
DATE:
VISIT TO PRAWN HATCHERY
AIM: To study the structure and functions of the prawn/shrimp hatcheries.
INTRODUCTION
Shrimp is a valuable aquatic food resource high in protein and commands good
export markets/ It has become the main target commodity for aqua farming in recent years.
Traditionally, shrimp fry are trapped and held in ponds and later collected by shrimp
gatherers for stocking in grow-out ponds. With increasing demand for shrimp, supply of
wild fry for the increasing number of shrimp farms has become insufficient and
inconsistent. The breakthrough in the completion of the life cycle of commercially
important shrimps in captivity, such as the tiger shrimp (Penaeus monodon), has greatly
enhanced mass production of shrimp fry under hatchery conditions. The excellent growth
performance of these hatchery-bred fry in grow-out ponds strongly shows that the shrimp
hatchery can answer the industry needs for ample supply of shrimp fry for farming.
Method
A field trip was made to Ramayapatnam village of Gudlur Mandal of Prakasm

district on 6-8-2016. We visited the Kings hatchery that is present on the bank of
Buckingham canal near Bay of Bengal. The equipotent and facilities that are there were
noted. The process of various operations in the farm on that were observed.
Report
Location:
This penaeid hatchery is located near the sea shore where clean water can be
pumped easily and economically. They are free from pollution that is away sources of
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Agricultural, Biological and Industrial wastes.
Climate:
This hatchery is e located in areas relatively dry from November to April and wet
during rest of the year. This type of climate will help in providing optimum temperature
(28 - 30 C)
Sea water quality: Sea water for hatchery is having the salinity range from 30 ppt to 35
ppt,..
Power supply:
Continuous power supply is present during the entire larval period for running Air
blower/aerator, pumps, lights and other domestic equipment used in the hatchery. They
have a stand by generator in case of power failure. That is 10 KVA3 phase generator
operated by 16 H.P. diesel motor.
Fresh water supply:

Continuous freshwater supply is there in the hatchery for lowering salinity when
acclimating post larvae, to reduce the salinity from 35 ppt to 30 ppt, for washing and for
the uses in the hatchery.
Transportation facilities:
The hatchery is connected with food transportation facilities with railway or road
for seed lifting and to transport of materials and other required things for hatchery.
Hatchery facilities and equipment:
This Prawn hatchery is having complete facilities and necessary equipment for successful
operation. It is having suitable tanks for larval and post larval rearing, Algal or
phytoplankton and Zooplankton culture, air and sea water supply systems.
TANKS
Tanks for rearing of larvae and post larvae: Larval tank capacity varies from 1 to 20 tones.
For economical operation, a larval rearing tank should have a water capacity of 3-5 tons,
while a post larval (nursery) rearing tank should held 6-10 tons, both at 1 m depth, one 3
ton larval tank can hold from 1,50,000 to 3,00,000 nauplii obtained from a single spawner.
Algal culture (or) phytoplankton tanks: Small and shallow tanks of not more than 1 ton
capacity and about 0.5 m deep are in use for algal or phytoplankton culture. This is because
adequate light is necessary for faster algal growth the tanks may also be mad of bamboo and
plastic materials.
Aeration supply:
semester
Aeration is there to provide oxygen in the culture water and to keep larvae and food
in suspension. It is supplied by an electric blower/a compressor, or a portable aerator.
Sea Water Supply:

Using a single suction line lay a few feet above the sea bed. The intake pipe opening
is fitted with screen to prevent fish and other unwanted organisms from being sucked in.
Pumped water directed to the hatchery where it is thoroughly filtered before use. This filtered
sea water is free from turbidity, debris and other undesirable marine organism. This filtered
water is pumped to the overhead tank. These overhead tanks serve as a desiltation tank PVC
or brass pipes used for the distribution of sea water from the overhead storage tank.
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Building: A concrete building for office and dwelling is present. Asbestos sheds are there
for hatchery.
Hatchery equipment: The basic equipment that are there in a hatchery are:
1. Refractometer/Hydrometer: Required for monitoring the salinity from time to time

in larval tanks and in the water storage tanks.
2. Thermometer: Thermometer is required for monitoring the temperature of the water

in the larval tanks.
3. Heamocytometer: Heamocytometer is required for counting algal cells.
4. Refrigerator: For storing stock cultures ofAlgal and other feeds for larval and post
larval stages.
5. Microscope: Required for monitoring the conditions of larval and feed density
6. Air Diffusers: Air diffuser stones are used for serration in the larval tanks.
7. Scoop nets: Used for scooping larval of fry from tanks or from harvesting
8. Harvest box: to use for harvest of fry (post larvae).

9. Drainers: Drainers various types and mesh sizes for draining the water, during the
water exchange.
10. Glass beaker: Beaker or any other transparent container of 200 ml to 1 it. for
counting, feeding and checking of the condition of larvae.
Method of brood stock selection:

Brood stock supply: It is ideal for hatcheries to be near the source of wild sprawners
and brood stock. Wild sprawners (mother Shrimps) is usually caught with a shrimp trawl,
trawling time is usually kept limited, so as to avoid stress on mother prawns. Only sprawners
with late maturing or mature ovaries are selected for hatchery. If wild sprawners are scarce,
the brood stock can be developed at the hatchery site by eye-stalk ablation techniques.
In the selection marine prawn brood stock, only gravid female (female prawn
carrying full of ripped eggs) will be selected, since mating already take place in the sea. The
brood stock selected by technicians from the wild catch, for which the following desirable
characteristics. The prawn should have a clear outline of the ovary when observed from the
back. The matured ovary can be seen as three knobs in the anterior part of the abdomen. The
ovary should extend until distal part of the abdomen. An ovary which does not reach the
distal part of the abdomen is considered to have spawned partially. The colour of the ovary is
in dark green/light yellow/light green, which differs from prawn to prawn. The spaawning
success of the middle size group (20 cm in length and 80 Gms in weight) is very good. 70
prawns of brood stock are put in 200m tank for spawning.
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Egg collection and cleaning:

After spawning the eggs have to be taken out from the spawning tank. At the same
time the eggs also cleaned by washing thoroughly, to remove the rose colour substance.
This rose cleaning material comes along with eggs during spawning. If proper cleaning is
not attended very soon the larvae get infection. The broodstocks may be checked after
spawning, dead females and sprawners should be removed. The aeration in the spawner
tank should be adjusted so as to weakly bubble the water. The egg collection time usually
occurred in nights though it depends on the spawning condition of the brood stock. The
number estimation after sucking up the eggs using an air hose, put this method needs
judgments by experience.
Hatching & identification of larvae:

Fertilized eggs are hatched into oval shape free swimming larvae known as Nauplius. One
middle size female releases 3, 00,000 eggs and percentage of hatching is estimated 80%.
Identification of larvae stages:

Prawn larvae moults repeatedly and metamorphose in the following manner.
1. The first larval stage, Nauplius stage undergo five mountings to reach Nauplii VI
Stage (NI to B VI) of course in some species it is different.
2. Zoea stage which second larval stage after completing Nauplius, has three sub
stages, Zoea Ito Zoea III, third stage
3. Mysis also has three sub stages, Mysis Ito III, and the last stage post larvae stage in
which there is no sub stages.
4. For the cycle from hatching to reach post larvae stage, it takes about 12 to 20 days.
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110
Difficulty in back-washing.
5,Mn ,SWaa
WM
2w u
Detail of layout, cross-sectional of 40-tons concrete nursery
TON FIBERGLASS TANK UTILIZED FOR ALGAE CULTURE
semester
111
A gravity water f i l ter .
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112
sum
p
PlT
Water storage and filtration tank.
Experiment No 21
Date:
EXPERIMENT ON DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND
Aim: To determine biochemical oxygen demand in the given water sample
INTRODUCTION
The biochemical oxygen demand determination is a chemical procedure for
semester
113
determining the amount of dissolved oxygen needed by aerobic organisms in a water body
to break the organic materials present in the given water sample at certain
temperature over a specific period of time.
BOD of water or polluted water is the amount of oxygen required for the
biological decomposition of dissolved organic matter to occur under standard condition at a
standardized time and temperature. Usually, the time is taken as 5 days and the
temperature is 20°C.
The test measures the molecular oxygen utilized during a specified incubation
period for the biochemical degradation of organic material (carbonaceous demand) and the
oxygen used to oxidize inorganic material such as sulfides and ferrous ion. It also
may measure the amount of oxygen used to oxidize reduced forms of nitrogen (nitrogenous
demand).
ENVIRONMENTAL
SIGNIFICANCE
BOD is the principle test to give an idea of the biodegradability of any sample and
strength of the waste. Hence the amount of pollution can be easily measured by it.
Efficiency of any treatment plant can be judged by considering influent BOD and the
effluent BOD and so also the organic loading on the unit. Application of the test to organic
waste discharges allows calculation of the effect of the discharges on the oxygen resources
of the receiving water. Data from BOD tests are used for the development of engineering
criteria for the design of wastewater treatment plants.
Ordinary domestic sewage may have a BOD of 200 mg/L. Any effluent to be discharged
into natural bodies of water should have BOD less than 30 mg/L.
This is important parameter to assess the pollution of surface waters and ground waters
where contamination occurred due to disposal of domestic and dustrial effluents.
The determination of BOD is used in studies to measure the self purification capacity of
streams and serves regulatory authorities as a means of checking on the quality of effluents
discharged to stream waters.
The determination of the BOD of wastes is useful in the design of treatment facilities.
It is the only parameter, to give an idea of the biodegradability of any sample and self
purification capacity of rivers and streams.
The BOD test is among the most important method in sanitary analysis to detenmine the
polluting power, or strength of sewage, industrial wastes or polluted water.
It serves as a measure of the amount of clean diluting water required for the successful
disposal of sewage by dilution.
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PRINCPLE
The sample is filled in an airtight bottle and incubated at specific temperature for 5
days. The dissolved oxygen (DO) content of the sample is determined before and after five
days of incubation at 20°C and the BOD is calculated from the difference between initial
and fmal M .
The initial DO is determined shortly after the dilution is made; all oxygen uptake
occurring after this measurement is included in the BOD measurement.
MATERIALS
REQUIRED
1. BOD Incubator
2. Burette & Burette stand
3. 300 ml glass stopper BOD bottles
4. 500 ml conical flask
6. Pipette bulb
7. 250 ml graduated cylinders
8. Wash bottle
CHEMICALS REQUIRED
1. Calcium Chloride
2. Magnesium Sulphate
3. Ferric Chloride
4. Di Potassium Hydrogen Phosphate
5. Potassium Di Hydrogen Phosphate
6. Di sodium hydrogen phosphate
7. Ammonium Chloride
8. Manganous sulphate
9. Potassium hydroxide
10. Potassium iodide
11. concentrated sulfuric acid
12. Starch indicator
13. Sodium thiosulphate
14. Distilled or Deionized
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PREPARATION OF REAGENT
1. Monogamous Sulphate Solution:
Dissolved 50 grams of Manganese Sulphate in !00 ml of distilled water.
2. Alkaline Iodide Solution:
Dissolved 50 grams of potassium hydroxide and 25 grams of potassium iodide in !00 ml
of distilled water.
3. 0.025 N sodium thiosulphate solution: Dissolved 6.21 grams of sodium
thiosulphate in !00 ml of distilled water.
4. Starch solution : one gram of soluble starch is dissolved in 100 ml of distilled water
and boiled for 5
minutes. Add few drops of formaldehyde as preservative.
5. Concentrated sulphuric acid.
6. Phosphate Buffer :
8.5 grams of Potassium dihydrogen phosphate , 21 grams of Di Potassium hydro
genphosphate, 33.4 grams of sodium dihydrogen phosphate and 1.7 grams of
Ammonium chloride is dissolved in 500 ml distilled water and that solution is made
Upto one liter
7. Calcium Chloride solution
Weighed accurately 27.5 g of anhydrous calcium chloride and dissolved it in distilled
Water and made up to a liter with distilled water.
8. Magnesium sulph ate solut ion:
Weighed accurately 22.5 g of magnesium sulphate and dissolved it in 1000 ml distilled
water.
9. Ferric Chloride solution
Weighed accurately 0.25 g Ferric chloride and dissolve it in 1000 ml distilled
water.
10. Sodium sulphite solution;
Dissolve d 1.575 grams of sodium sulphite in one liter of distilled water
11. Glucose glutamic acid solution: 150 mg of glucose and 150 mg of glutamic acid
are dissolved in one liter distilled water.
Procedure
Dilution Water
High quality organic free water is used for dilutionpurposes.
For the test we have taken one liter of organic free aerated distilled water, hence added one
ml each of the nutrients.
1. Added 1 ml calcium chloride solution

2. Added lmL magnesium sulphate solution
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116
3. Added 1 ml ferric chloride solution and

4. Added 1 ml phosphate buffer solution
5. This is the standard dilution water. Preparation of dilution water was done 3 days
before initiating BOD test to ensure that the BOD of the dilution water is less than
0.2 mg/L.
TESTING OF SAMPLE
1. Four 300 ml glass stoppered BOD bottles were taken (two for the sample and two for the
blank).
2. Added 10 ml of the sample to each of the two BOD bottles and the fill the remaining
quantity with the dilution water. i.e., we have diluted the sample 30 times.
3. The remaining two BOD bottles were kept as blank, to these bottles added dilution water
alone.
4. After the addition immediately placed the glass stopper over the BOD bottles and noted
down the numbers of the bottle for identification.
5. Now preserved one blank solution bottle and one sample solution bottle in a BOD incubator
at 20°C for five days.
6. The other two bottles (one blank and one sample) were analyzed immediately.
7. Avoided any kind of bubbling and trapping of air bubbles. - No bubbles!
8. Added 2mL of manganese sulfate to the BOD bottle by inserting the calibrated pipette just
below the surface of the liquid.
9. Added 2 ml of alkali-iodide reagent in the sane manner.
10. Allowed it to settle for sufficient time in order to react completely with oxygen.
11. When this flock has settled to the bottom, shaked the contents thoroughly by turning it
upside down.
12. Added 2 ml of concentrated sulfuric acid via a pipette held just above the surface of the
sample.
13. Carefully stopperd and inverted several times to dissolve the floc.
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117
14. 50 ml of this solution was taken into a conical flask.
15. Rinsed the burette with sodium thiosulphate and then fill it with sodium
thiosulphate. Fixed the burette to the stand.
16. Added 1 ml of starch solution. and continued the titration until the blue color
disappears to colorless.
17. Noted down the volume of sodium thiosulphate solution added , which gives the
D.O. in mg/L. repeated the titration for concordant values.
18. After five days, took out the bottles from the BOD incubator and analyzed the
sample and the blank for DO.
19. Added 2mL of manganese sulfate to the BOD bottle by inserting the calibrated
pipette just below the surface of the liquid.
20. Added 2 ml of alkali-iodide reagent in the same manner.
21. Allowed it to settle for sufficient time in order to react completely with oxygen.
22. When this floc has settled to the bottom, shaken the contents thoroughly by turning it
upside down.
23. Added 2 ml of concentrated sulfuric acid .

24. Carefully stoppered and inverted several times to dissolve the floc.
25. Titration started immediately after the transfer of the contents to Erlenmeyer flask.
26. Rinse the burette with sodium thiosulphate and then fill it with sodium
thiosulphate. Fixed the burette to the stand.
27. Measure out 50 ml of the solution from the bottle into an Erlenmeyer flask.
28. Added 1 ml of starch solution and continue the titration until the blue color
disappears to colorless.
29. Noted down the volume of sodium thiosulphate solution added, which gives the
D.O. in mg/L. Repeat the titration for concordant values.
CAL CULAT ION

For determining the Biochemical Oxygen Demand in the given water sample, the readings
should be tabulated.
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TABLE
Burette Volume of
Volume of (mu) Titrant Dissolved
Trial
Sample R di (ml) Oxygen
No.y
(ml) Initial Final (Na2S2O3 (mg/L)
cnLitinn iic l\
Blank
1.
2.
Blank
1.
2.
Burette Solution: Sodium Thiosulphate

Pipette Solution: Sample
Indicator: Starch
End point : Disappearance of blue color
Specimen Calculation:
Initial DO of the diluted sample, (DO)

= 7.9
DO at the end of 5 days for the diluted sample,(
D5)= 3.2 mu Blank correction = CO - C5, (BC )
= 0.2
Initial DO of the blank, (CO)
= 8.2
DO at the end of 5 days for the blank, C5
= 8.0
Biochemical Oxygen Demand= {DO- D5 - BC} x Volume of the diluted sample

Volume of sample taken
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Biochemical Oxygen Demand (mg/L) = (7 . 9 - 3 . 2 - 0 . 2 )x

200/10
4.5x200/10
9 0 mg/L
INTERPRETATION OF RESULTS
The BOD of the given sample of water =.
DISCUSSION
On the basis of the BOD values, the characteristics of the water and the biological activity of
the incubated micro flora can be determined. Effluent with high
BOD levels is discharged into a stream or river; it will accelerate bacterial growth in the river
and consume the oxygen levels in the river. The oxygen may diminish to
levels that are lethal for most fish and many aquatic insects. As the river re-aerates due to
atmospheric mixing and as algal photosynthesis adds oxygen to the water, the oxygen
levels will slowly increase downstream. The biological capacity of a sewage treatment plant
can be tested by comparing the BOD value of a known control solution with
the BOD derived from the treatment plant.
BOD detects only the destructible proportion of organic substances and as a general
principle is therefore lower than the COD value, which also includes inorganic materials
and those materials which cannot be biologically, oxidized.
Experiment No.22
Date:
EXPERIMENT ON DETERMINATION OF CHEMICAL OXYGEN DEMAND
Aim : To determine chemical oxygen demand in the given water sample
INTRODUCTION
The chemical oxygen demand (COD) test is commonly used to indirectly measure the
amount of organic compounds in water. Most applications of COD determine the amount of
organic pollutants found in surface water (e.g. lakes and rivers), making COD a useful
measure of water quality. It is expressed in milligrams per liter (mg/L), which indicates the
mass of oxygen consumed per liter of solution.COD is the measurement of the amount of
oxygen in water consumed for chemical oxidation of pollutants. COD determines the
quantity of oxygen required to oxidize the organic matter in water or waste water sample,
under specific conditions of oxidizing agent, temperature, and time. This method covers the
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determination of COD in ground and surface waters, domestic and industrial wastewaters.
The applicable range is 3-900 mg/L.
COD values are particularly important in the surveys designed to determine and control the
losses to sewer systems. The ratio of BOD to COD is useful to assess the amenability of
waste for biological treatment. Ratio of BOD to COD greater than or equal to 0.8 indicates
that wastewater highly polluted and amenable to the biological treatment. It is useful to assess
strength of wastes, which contain toxins and biologically resistant organic substances.
COD can be related to TOC, however, does not account for oxidation state of the organic
matter. BOD value is always lower than COD value. For domestic and some industrial
wastewater, COD value is about 2.5 times BOD value.
PRINCIPLE
The organic matter present in sample gets oxidized completely by potassium

dichromate (K2Cr2O7) in the presence of sulphuric acid (H2S04), silver sulphate (AgSO4)
and mercury sulphate (HgSO4) to produce C02 and H2O. The sample is refluxed with a
known amount of potassium dichromate (K2Cr2O7) in the sulphuric acid medium and the
excess potassium dichromate (K2Cr2O7) is determined by titration against ferrous
ammonium sulphate, using ferroin as an indicator. The dichromate consumed by the sample is
equivalent to the amount of 02 required to oxidize the organic matter.
MATERIALS REQUIRED APPARATUS REQUIRED
COD Digester
Burette & Burette stand
COD Vials with stand
250 ml conical flask (Erlenmeyer flask)
Pipettes
Pipette bulb
Tissue papers
Wash Bottle
CHEMICALS REQUIRED
Potassium dichromate
Sulfuric acid
Ferrous ammonium sulphate
Silver sulphate
Mercury sulphate
Ferroin indicator
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Organic free distilled water
Standard Potassium Dichromate Reagent - Digestion Solution

Weighed accurately 4.913 g of potassium dichromate .and t 33.3g of mercuric sulphate and
dissolved in 167 ml of concentrated sulphuric acid . This solution is made upto a liter with
distilled water.
Sulphuric Acid Reagent - Catalyst Solution
Weighed accurately 5.5 g silver sulphate crystals to a dry clean 1000 ml beaker. To this
carefully added about 500 ml of concentrated sulphuric acid and allowed to stand for 24
hours (so that the silver sulphate crystals dissolve completely).
Standard Ferrous Ammonium Sulphate solution

Weighed accurately 39.2g of ferrous ammonium sulphate crystals and dissolved it in
distilled water and made upto a liter with distilled water.
SAMPLE HANDLING AND PRESERVATION
1. Samples are collected in glass bottles. Use of plastic containers is permitted if it is

known that there is no organic contaminants present in it.
2. Biologically active samples should be tested as soon as possible. Samples containing

settleable material should be well mixed, preferably homogenized, to permit removal of
representative aliquots.
3. Samples should be preserved with sulphuric acid to a pH < 2 and maintained at 40 C

until analysis.
4. Do not allow the samples to freeze.
PRECAUTIONS
1. The following precautions should be observed while performing the experiment:

2. Chlorides are quantitatively oxidized by dichromate and represent a positive
interference. Mercuric sulfate is added to the digestion tubes to complex the
chlorides so that it does not interfere in the determination.
3. Nitrites also interfere in the determination of COD and hence during the
determination of samples with high concentration of nitrites, 120mg of sulphuric
acid is added to the potassium dichromate solution.
4. Traces of organic material either from the glassware or atmosphere may cause a
positive error. Extreme care should be exercised to avoid inclusion of organic
materials in the distilled water used for reagent preparation or sample dilution.
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PROCEDURE TESTING
OF SAMPLE
1. Took three COD vials with stopper (two for the sample and one for the blank).
2. Added 2.5 ml of the sample to each of the two COD vials and the remaining COD
vial is for blank; to this COD vial added distilled water.
3. Added 1.5 ml of potassium dichromate reagent - digestion solution to each of the

three COD vials.
4. Added 3.5 ml of sulphuric acid reagent - catalyst solution in the same manner.
CAUTION: COD vials are hot now.
1. Caped tubes tightly. Switch on the COD Digester and fix the temperature at 150° C
and set the time at 2 hours.
2. Placed the COD vials into a block digester at 150°C and heated for two hours.
3. The digester automatically switches off. Then remove the vials and allow it to cool
to the room temperature.
4. Transfer the contents of the blank vial to conical flask.
5. Added few drops of ferroin indicator. The solution becomes bluish green in colour.
6. Titrated it with the ferrous ammonium sulphate taken in the burette.

7. End point of the titration is the appearance of the reddish brown colour.
8. Noted down the volume of ferrous ammonium sulphate solution added for the blank
(A) is 14.1 ml.
9. Transferred the contents of the sample vial to conical flask.
10. Added few drops of ferroin indicator. The solution becomes green in colour.
11. Titrate it with the ferrous ammonium sulphate taken in the burette.
12. End point of the titration is the appearance of the reddish brown colour.
13. Noted down the volume of ferrous ammonium sulphate solution added for the
sample (B) is 13.2 ml.
CALCULATION
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123
For determining the Chemical Oxygen Demand in the given water sample, the readings
should be tabulated.
TABLE
Volume of Burette Reading (ml)

Sample (ml) Volume of 0.1 N
S1 No. Sample FAS (ml)
Initial Final
2.
3.
Burette Solution: Ferrous Ammonium Sulphate

Pipette Solution: Sample
Indicator: Ferroin Indicator
End point: Appearance of reddish brown color
1. For the blank titration the volume of sample taken is 2.5 ml.
2. Ferrous Ammonium Sulphate is taken in the burette.
3. The obtain reading is 14.1 ml. Similarly for sample one the volume of sample taken
is 2.5 ml.
4. Ferrous Ammonium Sulphate is taken in the burette
5. The initial reading is 0 ml and the final reading is 13.2 ml. Volume of Ferrous
Ammonium Sulphate consumed to get the end point is 13.2 mL.
6. For sample two the initial reading is 0 ml and the final reading is 13.2 ml.
7. The volume of Ferrous Ammonium Sulphate consumed to get the end point is 13.2
ml.
8. For sample 1 and 2 the reading as same so we can go for the calculations.
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Volume of Burette Reading (ml) Volume of 0.1 N

S1 No. Sample Sample (ml) Initial Final FAS (ml)
4. Blank 2.5 0 14.1 14.1
5. 2.5 0 13.2 13.2

Sample 1
6. Sample 2 2.5 0 13.2 13.2
Specimen Calculation:
Volume of Ferrous Ammonium sulphate for blank (A) = 14.1 ml

Volume of Ferrous Ammonium sulphate for Sample (B) = 13.2 ml
Normality of Ferrous Ammonium sulphate N = 0.1 N
Volume of Sample V = 2.5 ml
Chemical Oxygen Demand =
(A - B * N * 8 * 1000) Volume of sample taken
To convert the sample size from ml to L, multiply the result by 1,000 miL to convert the sample
size from ml to L.
Residual Chlorine ( m g ) = (14.1 - 13.2) x 0.1 x 8 x 1000/2.5 =

288 mg/L
INTERPRETATION OF RESULTS
The COD of the given sample of water = 288 mg/L.
DISCUSSION
Chemical oxygen demand does not differentiate between biologically available and inert
organic matter, and it is a measure of the total quantity of oxygen required to oxidize all organic
material into carbon dioxide and water. COD values are always greater than BOD values. For
domestic and some industrial wastewater COD is about 2.5 times BOD.
125
DETERMINATION OF POROSITY OF SOIL

AIM: To determine the bulk density and porosity of soil
INTRODUCTION
Soil porosity" refers to the amount of pore, or open space between soil particles. Pore
spaces may be formed due to the movement of roots, worms, and insects; expanding gases
trapped within these spaces by groundwater; and/or the dissolution of the soil parent material.
Soil texture can also affect soil porosity
There are three main soil textures: sand, silt, and clay. Sand particles have diameters
between .05 and 2.0 mm (visible to the naked eye) and are gritty to the touch. Silt is smooth and
slippery to the touch when wet and individual particles are between .002 and .05 mm in size
(much smaller than those of sand). Clay is less than .002 mm in size and is sticky when wet. The
differences in the size and shape of sand, silt, and clay influence the way the soil particles fit
together, and thus their porosity.
Soil porosity is important in aquaculture for many reasons. A primary reason is that soil pores
contain the groundwater that is medium for any animals. Another important aspect of soil
porosity concerns the oxygen found within these pore spaces. All animals need oxygen for
respiration, so a well-aerated soil is important for growing crops. Compaction by construction
equipment or our feet can decrease soil porosity and negatively impact the ability of soil to
provide oxygen and water.
Bulk density of mineral soils commonly ranges from 1.1 to 1.5 g/cm3 in surface horizons. It
increases with depth and tends to be high in sands and compacted pan horizons, and tends to be
low in soils with abundant organic matter. Tillage operations loosen soils and temporarily lower
bulk density, while compaction processes raise bulk density. High bulk densities correspond to
low porosity Natural soil-forming processes that increase aggregation reduce bulk density, but
excessive tillage and raindrop impact on bare soil destroy aggregation and increase bulk density.
. It differs from bulk density because the volume used does not include pore spaces.
Particle density = oven-dry soil weight / volume of soil solids
Particle density represents the average density of all the minerals composing the soil. For most
soils, this value is very near 2.65 g/cm3 because quartz has a density of 2.65 g/cm3 and quartz is
usually the dominant mineral. Particle density varies little between minerals and has little
practical significance except in the calculation of pore space.
PRINCIPLE:
Porosity can be calculated if bulk density and particle density are known. Bulk density is
soil mass divided by unit volume. In its natural state, a soil's volume includes solids and pores;
therefore, a sample must be taken without compaction or crumbling to correctly determine bulk
density.
Porosity is that portion of the soil volume occupied by pore spaces. This property does not have
126
to be measured directly since it can be calculated using values determined for bulk density and
particle density. Finding the ratio of bulk density to particle density and multiplying by 100
calculates the percent solid space, so subtracting it from 100 gives the % of soil volume that is
pore space.
REQUIRTEMENT
1. Measuring cylinders
2. Balance
3. Hot air oven
PROCEDURE
1. Collect sample from three different areas
2. Measure the volume of soil With measuring cylinder
3. Weigh the soil sample and record weight.
4. Dry the soil sample in Hot air oven
5. Again weigh and record dry weight of soil sample.
RESULTS
S.NO. Parameter VALUE
1 Volume of soil sample ( ml)
2 Wet weight of the soil (g)
3 Dry weight of the sample (g)
4 Bulk density
5 Particle density
CALCULATIONS
6 Porosity (%) 1. Bulk density = Oven dry soil weight

/ volume of soil solids and pores
2. Particle density= volumetric mass of the solid soil
3. % solid space = (bulk density / particle density) x 100
4. % porosity = 100 - (% solid space)
5. Porosity of Soil =
MODAL E CALCULATION OF POROSITY:
A 260 cm3 cylindrical container was used to collect an undisturbed soil sample. The container
and soil weighed 413 g when dried. When empty the container weighed 75 g. What is the bulk
density and porosity of the soil?
A. To determine bulk density:
Sample Volume = 260 cm3; Sample Weight = 413 - 75 = 338 g; Bulk density = 338 g/260 cm3=
1.3 g /cm3
B. To determine porosity:
127
Bulk density = 1.3 g /cm3;

Particle density = 2.65 g /cm3;
Porosity = 100 - (1.3/2.65 x 100) = 51%
CONCLUSION
Soil water and air occupy voids in the soil, called pore spaces. The pore system in soil
provides the conduits for air and water exchange and houses roots and microbes. Soil porosity is
the amount of pore volume (%age of pore space). A medium textured, well-aggregated soil
contains about 50% pore space and is in good condition for plant growth when the pores hold an
equal distribution of air and water. Pore size affects pore activity. Big pores, macropores,
facilitate free-water drainage, aeration, evaporation, and gas exchange. Mesopores, medium-size
pores, are essential to capillary water distribution, and micropores provide water storage sites.
Macropores are most prevalent in sandy soils and well-aggregated soils, but can be converted to
micropores by compaction. Medium-textured soils have an abundance of mesopores. Clays
promote aggregation but can also be readily compacted. Clays also increase water storage by
providing an abundance of micropores. Thus, texture and structure, plus the level of induced
compaction, are the main properties governing amount and type of pore space in the soil. Organic
matter affects porosity through its enhancement of soil aggregation.
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1

AQUACULTURE LABORATORY EXPERIMENTAL PROTOCALS

S.NO. NAME OF THE EXPERIMENT PAGE

DETERMINATION OF pH OF WATER SAMPLES

AIM: To determine the pH of the given water sample.

Determination of pH is one of the important objectives in biological treatment of

Switch on the ph meter

4.0 7.G 9.2

Calibrate the pH meter Calibrate the pH meter

Calibrate the pH meter

SAMPLE HANDLING AND PRESERVATION

The following precautions should be observed while performing the experiment:

1. Buffer Solution of pH 4.0

2. Buffer Solution of pH 7.0

d. Made up the volume to 100 ml using distilled water. 3.

Buffer Solution of pH 9.2

CALIBRATING THE INSTRUMENT

1. In a clean dry 100 ml beaker water sample was taken..

S.No Name of the sample pH value

The pH value of sample A is =

ESTIMATION OF SALT CONTENT BY USING SALINITY REFRACTOMETER

illuminator flap with calibration bimetallic

sample measuring shadow boundary

1. Water droplet should be completely dispersed on the prism

S.No Name of the Salinity (ppt)

1. Salinity of the A sample ppt ppt

The salinity Refractometer is very useful to determine salinity in field conditions.

DETERMINATION OF FREE CARBONDIAXIDE CONTENT IN WATER

Aim: To determination of free carbon dioxide in waste water

Free carbon dioxide in domestic water is of no significance because it appears to have no

1. Phenolphthalein indicator solution : 500 mg of phenolphthalein is dissolved in 10 ml ethanol

1. 50 ml of unfiltered water sample is taken in a measuring cylinder avoiding agitation

S.N Name of Volume of

sample volum readings Run (

volume in ml of 0.05 N Sodium hydroxide rundown X 1000. _

The free carbondioxide content in sample A - mg/L

DETERMINATION OF TUBIDITY OF WATER BY SECCHI'S DISC METHOD

Aim: To study turbidity of water samples by Secchi's Disc method

b. Planktons: A water body may be turbid due

body is rich in nutrients.

Secchi's Disc method

Preparation of Secchi's Disc:

1. Visited a nearby aquacultural pond.

DETERMINATION OF CARBONATES AND BICARBONATES

Aim: To determine the carbonate and bicarbonate content of water.

2 NaHCO3 + H2SO4 1'2SO4 + 2 H2O+ 2 CO2

3. Phenolphthalein (0.25%): Dissolved 25 mg of pure Phenolphthalein powder in 100 ml of

bicarbonates plus initial bicarbonates present in irrigation water.

Calculations: Carbonates (mill equivalents / liter)

In recalculating systems where photosynthesis is practically non-existent, a good buffering

DETERMINATION OF ALKALINITY OF WATER SAMPLES

Aim: To determine the Alkalinity of water samples

2 NaHCO3 + H2SO4 N~22SO4 + 2 H2O + 2 C02

2. Phenolphthalein indicator : Dissolve 1 gm of pure Phenolphthalein powder in 10 ml of ethyl

Total alkalinity = Z ml X 1000 ml of sample

S. Nam Samp Volume of Volume of

Initi Midd Fin

EXPERIMENT ON DETERMINATION OF CALCIUM HARDNESS

AIM: To determine the hardness of water

2. Ammonia Buffer: for 11.4 ml of ammonium hydroxide , 1.35 ml of ammonium chloride is

EXPERIMENT ON DETERMINATION OF RESIDUAL CHLORINE

Aim: To determine residual chlorine in the given water sample

Chlorine residuals determination is used to control chlorination of domestic and industrial

1. Burette & Burette Stand