«xM

or;ii AGE,
UACUL

I
Z MANAGEMENT
ORGANISED BY

DVANCED STUDIES IN MARI

UNE FISHERIES RESEARCH INSTITU
C O U N C I L OF AGRICULTURAL RESEARCH

HIN FROM
vm
l
 THE CENTRE OF ADVANCED STUDIES IN MARICULTURE was
started in 1979 at the Central Marine Fisheries Research Institute,
Cochin. This is one of the Sub-projects of the ICAR/UNDP
project on ' Post-graduate Agricultural Education and Research'.
The main objective of the CAS in Mariculture is to catalyse
research and education in maricujture which forms a definite
means and prospective sector to augment fish production of the
country. The main functions of the Centre are to :

— provide adequate facilities to carry out research of excellence

in mariculture/coastal aquaculture;

— improve the quality of post-graduate education in mari.

culture;

— make available the modern facilities, equipments and the

literature;

— enhance the competence of professional staff;

— develop linkages between the Centre and other Institutions

in the country and overseas;

— undertake collaboration programmes; and

— organise seminars and workshops.

Under the programmes of the Centre, Post-graduate courses

leading to M.Sc. (Mariculture) and Ph.D. are offered in
collaboration with the University of Cochin since 1980.
 WATER QUALITY MANAGEMENT
IN AQUACULTURE

CLAUDE E. BOYD
Department of Fisheries and Allied Aquacultures,
Auburn University, Alabama

. with assistance from

V. K. PILLAI
Central Marine Fisheries Research Institute,
Cochin 682.018, India

CMFRI SPECIAL PUBLICATION

Number 22

ISSUED ON THE OCCASION OF THE WORKSHOP ON

WATER QUALITY MANAGEMENT
IN AQUACULTURE
ORGANISED BY
THE CENTRE OF ADVANCED STUDIES IN MARICULTURE,
CENTRAL MARINE FISHERIES RESEARCH INSTITUTE
INDIAN COUNCIL Of AGRICULTURAL RESEARCH
HELD AT COCHIN FROM 10-13 DECEMBER 1984
 CONTENTS

PREFACE lit
INTRODUCTION vii

WATER QUALITY AND FISH PRODUCTION 1

FISH FEEDING AND WATER QUALITY 29

FERTILIZATION 33

LIMING 45

DISSOLVED OXYGBN AND AERATION 50

MISCELLANEOUS TREATMENTS AND CALCULATION OF

TREATMENT RATES 63
WATER ANALYSIS 71
 INTRODUCTION

Water quality includes all physical, chemical and biological

factors that influence the beneficial use of water. Where fish
culture is concerned, any characteristic of water that affects the
survival, reproduction, growth, production, or management of
fish in any way is a water quality variable. Obviously, there are
many water quality variables in pond fish culture. Fortunately,
only a few of these normally play an important role. These are
the variables that fish oulturist should concentrate on, and
attempt to control to some extent by management techniques.

All other things being equal, a pond with ' good' water
quality will produce more and healthier fish than a pond with
' poor' water quality. In this manual an attempt is made to
define ' good' water quality for fish culture. Information is also
presented which will help in determining the potential of a body
of water for producing fish, improving water quality, avoiding
stress-related fish disease and parasite problems, maintaining fish
for research purposes and ultimately producing more fish
per unit of surface area. The following discussion of water
quality is brief, but an attempt has been made to cover the most
important points. Chapter 1 is concerned with general aspects of
water quality that influence fish production. The influence of fish
feeding on water quality is covered in Chapter 2. Chapter 3
covers pond fertilization and Chapter 4 is concerned with
liming. Dissolved oxygen and aeration is treated in Chapter 5.
Chapter 6 deals with miscellaneous treatments and procedures
for computing doses of chemicals for ponds. Methods of water
analysis are presented in Chapter 7.

The information in this manual is essentially a summary of a

book on water quality in pond fish culture by Boyd, ' Water
Quality Management in Pond Fish Culture \ This book may be
consulted for more details and for additional references to the
literature on water quality in ponds. Most of the basis for the
book and this manual was research on warm water pond fish
culture in the Southern United States. However, all of the
principles and methods are general and applicable tofishculture
in other regions including culture of fish in brackish water and
sea water ponds.
 1
WATER QUALITY AND FISH PRODUCTION

1.1. Introduction
The material in this report explains the usual relationships
between water quality variables and fish production, setting forth,
where possible, ranges of desirable levels of the variables. In
addition, the management procedures recommended herein
usually will be effective in improving water quality. However,
because of unexplained reasons, effects of water quality on fish
and the effectiveness of management procedures may be quite
different from those reported here. Therefore, fish culturists
should not consider the information in this report as the final
answer to water quality problems, but merely as suggestions on
how to solve these problems. Coldwater fish will not be
considered, but coldwater fish generally demand water of much
better quality than do warmwater fish.

1.2. Temperature
Warmwater fish grow best at temperatures between 25°C and
32CC. Water temperatures are in this range the year
around at low altitudes in the tropics, but in temperate regions
water temperatures are too low in winter for rapid growth of
fish and fish food organisms. For this reason, management
procedures such as feeding and fertilizing are halted or reduced
in winter. Temperature has a pronounced effect on chemical and
biological processes. In general, rates of chemical and biological
reactions are doubled for every 10°C increase in temperature. This
means that aquatic organisms will use twice as much dissolved
oxygen at 3Q°C as at 20°C and chemical reactions will progress
twice as fast at 30°C as at 20°C. Therefore, dissolved oxygen
requirements of fish are more critical in warmwater than in cold
water. Chemical treatments of ponds also are affected by
temperature. In warm water, fertilizers dissolve faster, her-
bicides act quicker, rotenone degrades faster and the rate of
oxygen consumption by decaying manure is greater.

1
i
In ponds, heat enters at the surface and so surface waters get
heated faster than lower waters. Since the density of water
(weight per unit volume) decreases with increasing temperature
above 4°C, the surface waters may become so warm and light
that they do not mix with the cooler, heavier waters in lower layers.
The separation of pond water into distinct warm and cool layers
is called thermal stratification. The upper warm layer is called
the epilimaion and the lower cooler layer is known as the
hypolimnion. The layer of rapidly changing temperature between
the epilimnion and the hypolimnion is termed the thermocline.
The temperature profile for a thermally stratified pond is shown
in Fig. 1.1. In temperate regions large ponds may stratify in the
spring and remain stratified until fall. In small, shallow ponds

Dopth,
mtltrs

Ttmp»roture,*C

FIG. 1.1. A well developed pattern of thermal stratification in a fish

pond. The epilimnion, thermocline and hypolimnion are indicated.

in temperate regions and in tropical ponds, stratification often

exhibits a daily pattern. During the day the surface waters warm
and form a distinct layer. At night the surface waters cool to
 3
the same temperature as the lower waters and the two layers mix.
An extensive discussion on thermal stratification may be found in
any standard text on limnology.

In some ponds, the surface waters may reach temperatures of

35°C or more. This is above the optimum temperature for most
warmwater fish, but the fish may seek haven from the high
temperature in subsurface waters. Fish have poor tolerance to
sudden changes in temperature. Therefore, one should not
remove fish from water of one temperature and suddenly thrust
them into a water of appreciably higher or lower temperature.
Often, a sudden change in temperature of as little as 5°C will
stress or even kill fish. The effect is usually worse when moving
fish from cooler to warmer water. Since temperatures increase
with decreasing altitude, one must allow for temperature adjust-
ment when moving fish from high altitude to low altitude waters.
Fish readily tolerate gradual changes in temperature. For
example, one could raise temperature from 25°C to 32°C over
several hours without harming fish, but fish suddenly removed
from 25°C water and placed in water of 32° C might die.

1.3. Salinity
The term salinity refers to the total concentration of all
dissolved ions in a natural water expressed in milligrams per
litre. The osmotic pressure of solutions increases with increasing
salinity. Fish species differ in their osmotic pressure require-
ments, so the optimum salinity for fish culture differs to some
extent with species. Salinity information on some cultured
species of pond fish is presented in Table 1.1.

Fish are highly sensitive to sudden changes in salinity. Fish

living in water at one concentration of salinity should not
suddenly be placed in water with a much higher or lower salinity.
Small fish and fry of most species are more susceptible than
adult fish to sudden changes in salinity. Sodium chloride may
bs used to increase the salinity in fish holding facilities and even
in small experimental ponds. Conversely, salinity may be lowered
in small scale systems by the addition of water with low salinity.
Unfortunately, it is usually not practical to adjust the salinity of
4
larger fish culture systems, except in brackishwater ponds where
seawater may be introduced by gravity flow or tidal movement.

TABLE 1.1, Highest concentrations of salinity which permit

normal survival and growth of some cultured food fish

Species Salinity (mg/litre)

Catla catla (catla) . slightly brackishwater

Labeo rohita (roha) . slightly brackishwater
Ctenopharyngodon idella (grass carp) 12,000
Cyprinus carpio (common carp) 9,000
Hypophthalmichthys molitrix (silver carp) 8,000
Ictalurus punctatus (channel catfish) 11,000
Tilapia aurea 18,900
T. nilotica 24,000
T. mossambica 30,000
Mugil cephalus (grey mullet) 14,500
Chanos chanos (milkfish) 32,000

In practice, one is seldom able to measure concentrations of

all ions in water. However, the ability of water to conduct an
electrical current (conductivity) increases as salinity rises. A
conductivity meter may be used to measure conductivity and the
conductivity value allows an approximation of salinity. Many
conductivity meters have a scale for reading salinity directly.
Another method for obtaining the approximate salinity of a
water is to measure the total dissolved solids concentration. A
sample is filtered through a fine paper, a known volume is
evaporated and the residue remaining is weighed. The weight
of the residue in milligrams par litre is the total dissolved solids
concentrations and this closely approximates the salinity. In
brackishwater, salinity may be estimated from the chloride
concentration by the following equation :
Salinity in mg/litre = 30 + (1.8Q5) chloride in (mg/litre).
In practice, chloride concentrations (chlorinity) oan be measured
with refractometers or temperature corrected hydrometers.
The degree of salinity in water reflects geological and
hydrological conditions. Surface waters in areas of high rainfall
 5
where soils are continually leached usually have low salinity (10
to 250 mg/litre). In arid regions, evaporation exceeds precipita-
tion and salinity increases as a result of evaporation. Salinity
values in ponds of arid regions often range between 500 and
2,500 mg/litre and much higher values are often encountered.
Even in areas of high rainfall, ground water from wells may
sometimes have salinity values as high as those encountered in
surface waters of arid regions. Seawater has a high salinity
(35,000 mg/litre), so the salinity of braokishwater ponds reflects
the degree of dilution of seawater and freshwater. High rates of
evaporation in brackishwater ponds during periods of low rainfall
may cause them to become excessively saline. Salinities in
excess of 45,000 mg/litre are difficult for even marine species to
tolerate.

1.4. Turbidity and Colour

The term turbid indicates that a water contains suspended

material which interferes with the passage of light. In fish
ponds, turbidity which results from planktonic organisms is a
desirable trait, whereas that caused by suspended clay particles is
undesirable. Even with the latter conditions, the clay particles
are seldom abundant enough in water to directly harm fish. If
the pond receives runoff which carries heavy loads of silt and
clay, the silt settles over the pond bottom and smothers fish eggs
and fish food organisms. The clay particles which remain in
suspension restrict light penetration and limit the growth of plants.
A persistent clay turbidity which restricts visibility into the water
to 30 cm or less may prevent development of plankton blooms.
Methods of controlling clay turbidity will be discussed later.

Some ponds receive large inputs of vegetative matter from

their watershed. Extracts from this plant material (humates)
often impart colour to the water. Colour from vegetative
extracts often appears as a dark stain giving the water the
appearance of tea or weak coffee. Pond waters with high concen-
trations of humates are typically quite acid and have a low total
alkalinity. Although colour does not affect fish directly, it
restricts light penetration and reduces plant growth. Agricultural
6
limestone applications have been used to successfully remove
humates from natural waters.

1.5. Plankton
Plankton is comprised of all the microscopic organisms which
are suspended in water and includes small plants (phytoplankton),
small animals (zooplankton) and bacteria. When there is
enough plankton in the water to discolour it and make it appear
turbid, the water is said to contain a plankton «bloom.' The
phytoplankton uses inorganic salts, carbon dioxide, water and
sunlight to produce its own food. The zooplankton feeds on
living or dead plankton and other tiny particles or organic matter

FIG. 1.2. A representative food web in a Sunfish-bass pond.

Tilopio

FIG. 1.3. A representative food web in a pond used for Tilapia culture.

in the water. Bacteria utilize any type of dead organic matter

in the water for food. In fish culture systems wherefishare not
provided supplemental feed, plankton forms the most abundant
base of the food web. Examples of food webs in fish culture
systems are given in Figs. 1.2 and 1.3. Both food webs begin
with phytoplankton growth. In Fig. 1.2 there are several steps
before ending with largemouth bass, while in Fig. 1.3 the food
 7
web is simpler because the Tilapiafeed directly on plankton. Since
each step in the food web is rather inefficient, a fish culture
system with a more direct food web will produce a greater weight
offishper unit area. For example, during a 6 month period, the
SunSsh-bass culture might produce 200 kg fish/ha while the
Tilapia culture could easily produce 1,000 kg/ha.

Sunfisn
production,
kg./ho

600

400

200

0 j i I i I 1.. I —I.
0 5 10 15 20 25 30 35
Particulate organic matter, mg/littr

Fio. 1.4. Plankton production (particulate organic matter) and Sunfisn

production in ponds.

Because plankton is at the base of the food web, thera is a

close relationship bat ween plankton abundance and fish produc-
tion (Fig. 1.4). In addition to encouragingfishgrowth, plankton
makes water turbid and prevents the growth of undesirable
aquatic weeds through shading. Despite the benefits of plankton
blooms infishponds, more plankton can sometimes be produced
than can be utilized by the fish for growth. Heavy plankton
8

blooms usually contain large numbers of blue-green algae which

can form scums at the surface. These scums absorb heat during
the day and cause shallow thermal stratification. During the
night, heavy plankton blooms consume large amount of dissolved
oxygen and may cause oxygen depletion bsfore the next morning.
Scums of plankton may suddenly die, decompose and cause
oxygen depletion. Relationships between plankton and dissolved
oxygen will be treated more thoroughly later. In addition to
causing dissolved oxygen problems, organisms in heavy plankton
blooms often produce substances which impart a strong off-flavour
tofishflesh.
There are several techniques for measuring the abundance or
metabolic activity of the phytoplankton. The most popular are
chlorophyll a determinations and measurements of primary
productivity. A method for chlorophyll a is provided later.
The total abundance of plankton is often ascertained from
particulate organic matter analysis. Unfortunately, these
techniques are too tedious for use in practical fish culture. The
most practical technique for use in ponds which do not contain
appreciable clay turbidity is to measure the Secchi disc visibility.
Details for making Secchi disc measurements will be given later,
but for now it will suffice to state that the Secchi disc visibility is
the depth at which a disc 20 cm in diameter with alternative
black and white quardrants disappears from view. There is a
high correlation between Secchi disc visibility and plankton
abundance, as illustrated in Fig. 1.5(1)*. It is impossible to
establish an ideal plankton turbidity for fish culture. However,
a Secchi disc visibility in the 30 to 60 cm range is generally
adequate for good fish production and for shading underwater
weeds. As Secchi disc visibilities decrease below 30 cm, there
is an increase in the frequency of dissolved oxygen problems.
At values above 60 cm, light penetrates to greater depths
encouraging underwater macrophyte growth and there is less
plankton to serve as food for fish or fish food organisms.

Plankton communities are constantly changing in species

composition and in total abundance. This results in corresponding

* Number in parentheses indicates the reference in the Bibliography at the

end of each chapter.
 9
fluctuations in Secchi disc visibility and in the appearance of
pond water. These changes in plankton communities may be
disconcerting to the fish culturist. However, unless plankton
becomes so dense that dissolved oxygen problems occur or so thin
as to encourage underwater weeds, the changes do not affect fish
Particulate
organic maiier,
mg/li»er

'Secchi disk visibility, miters

Fio. 1.5. Relationship between plankton abundance (particulate

organic matter) and Secchi disc visibility in fish ponds.
production appreciably. By monitoring Secchi disc visibility on
a regular schedule (once or twice weekly) and observing the
appearance of the water, the fish culturist can obtain information
on the continuing condition of the plankton community in a pond
and on the supply of fish food organisms.
10
The ability of water to produce plankton depends on many
factors, but the most important is usually the availability of
inorganic nutrients for phytoplankton growth. Essential elements
for phytoplankton growth include carbon, oxygen, hydrogen,
phosphorus, nitrogen, sulphur, potassium, sodium, calcium,
magnesium, iron, manganese, copper, zinc, boron, cobalt,
chloride and possibly others. Phosphorus is most often the
element regulating phytoplankton growth in ponds. The
addition of phosphate fertilizer will cause an increase in plankton
production and an increase in fish production in most ponds.
Inadequate supplies of nitrogen, potassium and carbon also
limit phytoplankton in some ponds. Nitrogen may be limiting in
brackishwater and seawater ponds.

Even though the basic fertility of ponds differs greatly depend-

ing on the management and soils of their watersheds, the level of
plankton production in most ponds can be raised within the range
of plankton production needed for good fish production. Inorganic
fertilizers may be added to ponds with low basic fertility to
increase plankton production. In some ponds, both lime and
fertilizer application may be required to increase plankton
production. Manures also increase plankton production.

1.6. Dissolved Oxygen

Dissolved oxygen is probably the most critical water quality
variable in fish culture, so the fish farmer should be familar
with the dynamics of dissolved oxygen concentrations in ponds.
The atmosphere is a vast reservoir of oxygen, but atmospheric
oxygen is only slightly soluble in water. The solubility of oxygen
in freshwater at different temperatures and at standard sea level
atmospheric prsssure is given in Table 1-2. From this Table, it is
readily apparent that the solubility of oxygen in water decreases
as the temperature increases. When water contains a dissolved
oxygen concentration equal to the solubility of oxygen in water
at the existing temperature, the water is said to be saturated
with dissolved oxygen. If water contains more dissolved oxygen
than it should for the particular temperature, it is supersaturated.
Water may also contain less dissolved oxygen than the saturation
value. The solubility of dissolved oxygen decreases with
 11

TABLE 1.2. Solubility of dissolved oxygen in pure water at

standard sea level Atmospheric pressure (1 atmosphere)

°C mg/litre °C mg/litre °C mg/litre

0 . 14.16 12 .. 10.43 24 8.25

1 13.77 13 10.20 25 8.11
2 13.40 14 9.98 26 7.99
3 13.05 15 9.76 27 7.86
4 12.70 16 9.56 28 7.75
5 . 12.37 17 9.37 29 7.64
6 12.06 18 9.18 30 7.53
7 11.76 19 9.01 31 7.42
8 . 11.47 20 8.84 32 7.32
9 . 11.19 21 .. 8.68 33 7.22
10 . 10.92 22 8.53 34 7.13
11 10.67 23 8.38 35 7.04

decreasing atmospherio pressure (barometric pressure). For

example, the solubility of oxygen in water at 25°C differs as
follows with altitude (given in milligrams per litre at specified
altitudes): 0 m, 8 4 mg/litre; 500 m, 7-9 mg/litre; 1,000 m,
7-4 mg/litre; 1,500 mg, 7-0 mg/litre; 2,000 m, 6-6 mg/litre;
2,500 m, 6-2 mg/litre; 3,000 m, 5-8 mg/litre. The solubility of
oxygen in water also decreases as salinity increases. At
temperatures of 20 to 35°C, the solubility of dissolved oxygen
decreases by about 0*008 mg/litre for each 210 mg/litre increase in
salinity. Thus, salinity is not an important factor regulating the
concentration of oxygen in freshwater. Sea water has a high
salinity and dissolved oxygen concentrations at saturation
(Table 1*3) are considerably lower than for freshwater.

TJven though dissolved oxygen will diffuse into water, its rate
of diffusion is quite slow. Therefore, photosynthesis by phyto-
plankton is the primary source of dissolved oxygen in a fish
culture system. Fish culturists are often concerned with the
rate at which dissolved oxygen is removed from the water. The
primary losses of dissolved oxygen from a pond include respiration
by the plankton (phytoplankton include), respiration by fishes,
respiration by benthic organisms (organisms living in or attached
to the mud) and diffusion of oxygen into the air (7,10). The
12

TABLE 1.3. Solubility of dissolved oxygen in sea water at

standard sea level atmospheric pressure (1 Atmosphere)

°C mg/litre °.C mg/litre °C mg/litre

0 11.41 12 8.58 24 6.83

1 11.11 13 8.41 25 6.71
2 10.83 14 8.24 26 6.60
3 10.56 15 8.07 27 6.49
4 10.30 16 7.91 28 6.38
5 10.05 17 7.78 29 6.28
6 9.82 18 7.61 30 6.18
7 9.59 19 7.47 31 6.08
8 9.37 20 7.33 32 5.99
9 9.16 21 7.20 33 5.90
10 8.96 22 7.07 34 5.81
11 8.77 23 6.95 35 5.72

gains and losses of dissolved oxygen in a pond are summarised in

Table 1*4 along with some values representing the usual
magnitudes of daily gains and losses. It is readily apparent
that plankton and fish respiration cause the major losses of
dissolved oxygen and that photosynthesis is the largest source.
Diffusion of oxygen into ponds only occurs when waters are
below saturation and diffusion of oxygen out of ponds only occurs

TABLE 1.4. Ranges of expected gains and losses of dissolved

oxygen caused by different processes in fish ponds, for
ponds of 1.0 to 1.5 metres average depth

Process Range
mg/litre

Gains
Photosynthesis by phytoplankton 5 to 20
Diffusion 1 to 5
Losses
Plankton respiration .. 5 to 15
Fish respiration .. 2 to 6
Respiration by organisms in mud 1 to 3
Diffusion 1 to 5
 1*
when waters are supersaturated. The larger the difference between
the dissolved oxygen concentration in the pond water and the
concentration of dissolved oxygen at saturation, the greater is the
rate of diffusion. Wind and wave action also favour diffusion.
In a fish culture system, more oxygen must enter or be
produced in the water by plankton than is used by the organisms
or dissolved oxygen depletion will occur. Since nutrients are
normally abundant in well managed fish ponds, light is often the
primary factor regulating photosynthesis by phytoplankton.
Light rapidly decreases in intensity as it passes through water.
This is true even in pure water, but the decrease is even faster in
fish ponds because the planktonic organisms and other suspended
and dissolved substances reflect and absorb light. Therefore,
the rate of oxygen production by phytoplankton decreases with
depth and below a certain depth, no more oxygen is produced.
Since oxygen is continually used by the pond biota and only
produced during daylight hours by the phytoplankton, there is a
depth at which dissolved oxygen production by the
phytoplankton and that entering by diffusion just equal the
combined utilization of dissolved oxygen by pond life. Below
this depth in stratified ponds the water will contain no dissolved
oxygen. The stratification of dissolved oxygen in ponds usually
corresponds closely to thermal stratification (3). The epilimnion
contains dissolved oxygen and the hypolimnion is depleted of
dissolved oxygen. As with thermal stratification daily dissolved
oxygen stratification may occur in small shallow ponds.
Obviously, the depth to which light intensity is great enough
for adequate photosynthesis to provide surplus dissolved oxygen
is related to plankton density. Photosynthesis decreases with
decreasing light intensity and as plankton becomes more abundant,
the rate of oxygen consumption by the plankton community
increases. When plankton abundance is great, dissolved oxygen
production is extremely high near the surface. Because of
shading, the rate of oxygen production will decrease rapidly with
depth and only a thin layer of surface water, often less than 1 m
will contain appreciable dissolved oxygen. In ponds where
plankton is less abundant, rates of dissolved oxygen production
are not as high within the illuminated layer of water, but there
will be appreciable oxygen production and surplus dissolved
14
oxygen at greater depths than in ponds with greater plankton
turbidity. The influence of plankton turbidity on the depth
distribution of dissolved oxygen in ponds is illustrated in Fig. 1.6.
As a general rule, most ponds will contain enough dissolved
oxygen to support fish to a depth of at least two or three times
the Secchi disc visibility.

e 12 i6 20 24
Di»»olv«d oiyg«n,mq/lit«r

FIG. 1.6. Dissolved oxygen conce trations in the afternoon at different

depths in ponds with different densities of plankton.

There is also a marked fluctuation in dissolved oxygen

concentration during a 24 hour period in ponds. Concentrations
of dissolved oxygen are lowest in the early morning just after
sunrise, increase during daylight hours to a maximum in late
afternoon and decrease again during the night. The magnitude
of fluctuation is greatest in ponds with heavy plankton blooms
and least in ponds with low plankton abundance. Daily
fluctuations of dissolved oxygen concentrations in ponds with
different plankton densities are depicted in Fig. 1.7. In ponds
with extremely dense plankton blooms, dissolved oxygen concen-
trations will often be below 2 mg/litre in early morning. Concen-
trations are particularly low during periods of cloudy weather
 15
Dissolved
oxygen,
mg/liter
20
heavy plankton
- uT bloom

16
>^_,\ moderate ptanMon

1
^*°->>X. bloom
\Z
»
^ v
light plankton
bloom

7i i 1 U— 1—
6o.m Noon 6 p.m. 12 p.m. 6am.
Time of day

FIG. 1.7. Daily fluctuations in dissolved oxygen concentrations of surface

water in ponds with different densities of plankton.
Dissolved
oiygen,
mg/liter

6o.m. 6o.m. 6o.m. 6am.

Time of day

FIG. 1.8. Influence of cloudy weather on dissolved oxygen concentrations

in fish ponds.
16*
(5, 12). The production of oxygen on a cloudy day is less than
on a clear or partly cloudy day, so dissolved oxygen concentrations
do not increase to usual afternoon levels. This results in lower
than usual dissolved oxygen concentrations the following morning.
Extended periods of cloudy weather may result in dangerously
low dissolved oxygen concentrations even in ponds with
moderately heavy plankton blooms. The influence of cloudy
weather on dissolved oxygen concentrations is illustrated in
Fig. 1-8.
Total
phytoplankton,
thousands/ml

I I < I I I I I I I I I I ! I I > I I I •
18 22 26 30 4 8 12
April May

FIG. 1.9. Decline in phytoplankton following a phytoplankton die-off

in a fish pond. The die-off began on April 29.
 1)
In ponds with heavy plankton blooms, scums of algae often
form at the surfaoe. Occasionally, the algae in these scums will
suddenly die and their decomposition will result in depletion of
dissolved oxygen (2, 6, 12). For example, Fig. 1-9 illustrates the
sudden death of phytoplankton in a fish pond. The dissolved
•Dissolved
ouygen,
mg/liter

Fio. 1.10. Concentrations of dissolved oxygen before and after a phyto-

plankton die-off in a fish pond. The phytoplankton began dying on
April 29.

oxygen concentration quickly dropped below a detectable level

(Fig. 1.10). Dissolved oxygen concentrations did not return to
normal levels until a new phytoplankton community was
established (Figs. 1.9 and 1.10). Phytoplankton die-offs usually
occur during calm, clear, warm weather. Oae can recognize a
die-off because the algae scum deteriorates and the water fakes
an a brown or gray appearance.
2
18
Winds or heavy, cold rains may break up thermal stratification
in ponds (12), causing complete mixing ('overturn') of oxygen-
less waters of the hypolimnion and the oxygenated water of the
epilimnion. If the pond contains a large volume of oxygenless
water, oxygen depletion may result.

Fish require adequate concentrations of dissolved oxygen for

survival and growth. The minimum concentration for fish
survival varies with time of exposure. A fish may tolerate a
particularly low concentration of dissolved oxygen for a few hours
without ill effect, but will die if exposed to this same
concentration for several days. The concentration of dissolved
oxygen tolerated by pond fishes is illustrated in Fig. 1.11, with
additional data on oxygen requirements presented in Table 1.5.
Low dissolved oxygen concentrations adversely affect fish even at
levels which do not cause mortality, making them more succepti-
ble to parasites and diseases (9). In addition, fish do not feed
or grow as well when dissolved oxygen concentrations remain
continuously balow 4 or 5 mg/litre (5). Daily fluctuations of
dissolved oxygen in fish ponds apparently have little effect on
feeding and growth as long as the minimum dissolved oxygen
concentration for the day does not drop below 1 or 2 mg/litre in
the early morning and then rises near saturation within a few
hours after sunrise. If dissolved oxygen concentrations remain
at lessthan 3 or 4 mg/litre for prolonged periods, fish cease to feed
or grow well.

TABLE 1.5. Reported lethal concentrations of dissolved oxygen

for selected species of pond fish

Species Lethal level

(rag/litre)
Carassius auratus (goldfish) 0.1 to 2.0
Catla catla (catla) 0.7
Cirrhina mrigala (rarigal) 0.7
Ctenopharyngodon idella (grass carp) 0.2 to 0.6
Cyprinus carpio (common carp) 0.2 to 0.8
Hypophthalmichthys molltrix (silver carp) 0.3 to 1.1
Labeo rohita (rohu) 0.7
Ictalurus punctatus (channel catfish) 0.8 to 2.0
Lepomis macrochirus (bluegill) 0.5 to 3.1
Micropterus salmoides (largemouth bass) 0.9 to 3.1
 49
1.7. pit
The pH is a measure of the hydrogen ion concentration and
indicates whether the water is acidic or basic in reaction. ThepH
scale ranges from 0 to 14, with pH 7 being the neutral point. Thus,
a water of pH of 7 is neither acidic nor basic, while a water with
pH below 7 is acidic and one with a pH above 7 is basic. The
greater the departure from pH 7, the more acidic or basic a water'.

0 small fish survive

short exposure
0.3
) lethal if exposure
J prolonged
1.0

0)

2,0
E
c"
V
>. 3.0 ^fish survive but growth
O
X>
slow for prolonged
4)
_> exposure
O
M
M
4,0

5.0

1 desirable range

Fro. 1.11. Effects of dissolved oxygen concentrations on pond fish.

The pH of natural waters is greatly influenced by the concen-

tration of carbon dioxide, an acidic substance. Phyfoplankton
and other aquatic vegetation remove carbon dioxide from the
water during photosynthesis, so the pH of a body of water rises
during the day and decreases during the night (Fig. 1.12).
Waters with low alkalinity often have pH values of 6 to 7*5
before daybreak, but when phytoplankton growth is heavy,
afternoon pH values may rise to 10 or even higher (11).
Fluctuations in pH are not as great in water with higher total
alkalinity where pH values normally range from 7.5 or 8 at
daybreak or 9 or 10 during the afternoon. In soms water with
extremely high total alkalinity and particularly in water with

PH
10-

7
^o.m. Noon 6p.m. 12 p.m. 6 am.
Time of day
Fro. 1.12. Dailyfluctuationsin pH in a fish culture pond.

high total alkalinity and low total hardness, pH values may

rise above 11 during periods of rapid photosynthesis (11).
Obviously, pH measurements should be made in the early
morning and again in the afternoon to assess the typical pH
pattern for a pond. Waters with pH values of above 6.5 to 9 at
daybreak are considered best for fish production. Some ponds
which receive drainage from acid soils or swamps may be too acidic
for fish production. Waters with extremely high total alkalinity
may have pH values too high for fish culture. Methods for
increasing or decreasing the pH of pond water will be given
later.
 21
The acid and alkaline death points for pond fish arc approxi-
mately pH 4 and pH 11 respectively (11). Even though fish,
may survive, production will be poor is pond with early morning
pH values between 4 and 6 and between 9 and 10 (Fig. 1.13). The
afternoon pHin many fish culture systems rises to 9 or 10 for
short periods without adverse effect on fish.

4 ocid deoth point - i

5 :} no reproduction
> slow growth

6
7
x desirable ronge for fish
* 8 production

9
1J#

10

II alkaline deoth point

Fio. 1.13. Effect of pH on pond fish.

|.8, Carbon dioxide

High concentrations of carbon dioxide can be tolerated by
fish, although fish avoid levels as low as 5 mg/litre. Most
species will survive in waters containing up to 60 mg/litre
carbon dioxide, provided concentrations are high (8). When
dissolved oxygen conosstrations are low, the presence of
appreciable carbon dioxide hinders the uptake of oxygen by the
fish. Unfortunately, carbon dioxide concentrations are normally
quite high when dissolved oxygen concentrations are low. This
results because carbon dioxide is released in respiration and
utilized in photosynthesis. When dissolved oxygen is low,
photosynthesis is not proceeding rapidly. Therefore, carbon
dioxide concentrations rise because carbon dioxide released by
22
respiration is not absorbed by phytoplankton for use in photo-
synthesis. Because of the relationship of carbon dioxide to
respiration and photosynthesis, carbon dioxide concentrations
usually increase during the night and decrease during the day.
Particularly high concentrations of carbon dioxide occur in ponds
after phytoplankton die-offs, after destruction of thermal
stratification and during cloudy weather.
The influence of photosynthesis on pH is caused by carbon
dioxide uptake according to the following equation :
2HCOs~ . CO, + CO/" + HaO.
As plants remove carbon dioxide for use in phytosynthesis,
carbonate accumulates in the water. Carbonate hydrolyzes as
follows:
C08*~ + HaO « HCOs- + OH".
The accumulation of hydroxide causes the pH to rise. At night,
plants quit removing carbon dioxide and carbon dioxide from
respiratory processes accumulates in the water. This causes the
pH to drop because carbon dioxide, carbonate and water react
to form bicarbonate. In other words, the reactions shown above
go to the right during the day, but at night they go to the left.

1.9. Ammonia
Ammonia reaches pond water as a product of fish metabolism
and decomposition of organic matter by bacteria. In water,
ammonia nitrogen occurs in two forms, un-ionized ammonia and
ammonium ion, in a pH and temperature dependent equilibrium :
NH, + H,0 = NH4+ + OH".
As pH rises, un-ionized ammonia (NH,) increases relative to
ammonium ion. Temperature also causes an increase in the
proportion of un-ionized ammonia, but the effect of increasing
temperature is less than that of increasing pH. Percentages of
un-ionized ammonia at different temperatures and pH values are
given in Table 1.6. Analytical procedures for ammonia measure
both un-ionized ammonia and ammonium ion (total ammonia
nitrogen). The percentage un-ionized ammonia for the appro-
priate temperature and pH may be multiplied by the total
ammonia nitrogen concentration to estimate the concentration of
 23
un-ionized ammonia. For example, suppose the pH is 9.0,
temperature is 28°C and total ammonia- nitrogen is 1.0 nig/litre.
The percentage un-ionized ammonia is41.2% atpH 9 and 28 C C;
the un-ionized ammonia concentration is 1 x 0.412 =»
0.41 mg/litre. The answer is in terms of ammonia-nitrogen; to
convert to ammonia, multiple by the ratio 17/14—the molecular
weight of ammonia to the atomic weight of nitrogen.

TABLE 1.7. Percentage un-ionized ammonia in aqueous solution

at different pH values and temperatures

Temperature (*Q
PH 16 18 20 - 22 24 26 28 30 32

7.0 .. 0.30 0.34 0.40 0.46 0.52 0.60 0.70 0.81 C.95
7.2 .. 0.47 0.54 0.63 0.72 0.82 0.95 1.10 1.27 1.50'
7.4 .. 0.74 0.86 0.99 1.14 1.30 1.50 1.73 2.00 2.36
7.6 .. 1.17 1.35 1.56 1.79 2.05 2.35 2.72 3.13 3.69
7.8 . 1.84 2.12 2.45 2.80 3.21 3.68 4.24 4.88 5.72
8.0 .. 2.88 3.32 3.83 4.37 4.99 5.71 6.55 7.52 8.77
8.2 .. 4.49 5.16 5.94 6.76 7.68 8.75 10.00 11.41 13.22
8.4 .. 6.93 7.94 9.09 10.30 11.65 13.20 14.98 16.96 19.46
8.6 .. 10.56 12.03 13.68 15.40 17.28 19.42 21.83 24.45 27.68
8.8 .. 15.76 17.82 20.08 22.38 24.88 27.64 30.68 33.90 37.76
9.0 .. 22.87 25.57 28.47 31.37 34.42 37.71 41.23 44.84 49.02
9.2 .. 31.97 35.25 38.69 42.01 45.41 48.96 52.65 56.30 60.38
9.4 .. 42.68 46.32 50.00 53.45 56.86 60.33 63.79 67.12 70.72
9.6 .. 54.14 57.77 61.31 64.54 67.63 70.67 73.63 76.39 79.29
9.8 .. 65.17 68.43 71.53 74.25 76,81 79.25 81,57 83.68 85.85
10.0 .. 74.78 77.46 79.92 82.05 84.00 85.82 87.52 S9.05 90.58
10.2 .. 82.45 84.48 86.32 87.87 89.27 90.56 91.75 92.80 93.84

As ammonia concentrations increase in the water, ammonia

excretion by fish diminishes and levels of ammonia in blood and
other tissue increases. The result is an elevation in blood pH
and adverse effects on enzyme-catalyzed reactions and membrane
stability. The permeability of the fish by water is affected and
internal ion concentrations increase. Ammonia increases oxygen
consumption by tissues, damages gills and reduces the ability of
blood to transport oxygen. Histological changes occur in the
gills, kidneys, spleen, thyroid and blood of fish exposed to
sublethal ooncentrations of ammonia. Disease susceptibility also
increases in fish exposed to sublethal concentrations of ammonia.
24
The tolerance of fish to ammonia varies with species,
physiological condition and environmental conditions. Lethal
concentrations for short-term exposure (24 to 72 hours) are
bstween 0.4 and 2 mg/litre (S). Sublethal effects (histological
changes) have been attributed to ammonia concentrations as low
as 0.01 mg/litte. Ammonia concentrations as low as 0.06 mg/
litre have slightly reduced growth in channel catfish; 0.52 mg/
litre caused a 50% reduction in growth in this species; no growth
occurred at 0.97 mg/litre.
It is difficult to evaluate ammonia concentrations in ponds
because of the daily cycle in pH. Un-ionized ammonia may be
quite high in the afternoon and negligible during the night.

1.10. Nitrite
When nitrite is absorbed by fish, it reacts with haemoglobin to
form methemoglobin. Because methemoglobin is not an effective
oxygen carrier, continued absorption of nitrite can lead to
hypoxia and cyanosis. Blood containing methemoglobin is
brown, so nitrite poisoning in fish is frequently called brown-
blood disease.
The sources of nitrite in fish ponds have not been definitely
identified. However, the most likely source is the reduction of
nitrate to nitrite in anaerobic muds. Regardless of the source,
ponds occasionally contain nitrite concentrations of 0.5 to 10
mg/litre.
It is difficult to establish a lethal threshold value for nitrite,
for its toxicity is related to water quality. Chloride and appar-
ently calcium ions reduce the toxicity of nitrite to fish. As long
as the molar ratio of chloride to nitrite does not drop below 3,
channel catfish are not harmed by nitrite. Hence, sodium
chloride application may be used to prevent brown-blood disease
when nitrite concentrations are high. Calcium chloride is more
effective than sodium chloride in alleviating nitrite toxicity in
trout and salmon. Thus, the toxic level for nitrite varies with
chloride and possibly calcium concentrations. Of course, a given
level of nitrite in a particular water results in a specific percent-
age of methemoglobin in fish. However,' the percentage, of
 15
methemoglobin that is necessary to harm fish will differ wjth
dissolved oxygen concentration. There is also evidence that
the toxicity of nitrite increases with decreasing pH. Nilritfi
concentrations above 1 mg/litre in pond water are usually
considered undesirable unless chloride concentrations are several
milligrams per litre.

Ml. Hydrogen sulphide

Under anaerobic conditions, certain heterotrophic bacteria
can use sulfate and other oxidiaied sulfur compounds as terminal
electron acceptors in metabolism and excrete sulphide as illustrated
b*low *
S0 4 »- + 8H+ + S*~ + 4HaO.
The sulphide excreted is an ionization product of hydrogen,
sulphide and participates in the following equilibria:
H2S = HS + H+;
HSB=S»~ + H~.

The pH regulates the distribution of total reduced sulfijr

among its forms. Un-ionized hydrogen sulfide is toxic to fish)
but the ions resulting from its dissociation are not appreciably
toxic. Analytical procedures measure total sulphide; values given
below show the percentage of un-ionized hydrogen sulfide aj
different pH values:
pH Hydrogen sul
5.0 99.0
5.5 97.0
6.0 91.1
6.5 76.4
7.0 50.6
7.5 24.4
80 9.3
8.5 3.1
9.0 1.0

Concentrations of hydrogen sulphide of 001 to 0.05 mg/litre

are lethal to fish and any detectable concentration of bydropep
26

sulphide in pond water is considered undesirable. Obviously, if

the pH of a water can be increased by liming, the toxicity of
hydrogen sulphide will decrease.

1.12. Total alkalinity and total hardness

The term total alkalinity refers to the total concentration of

bases in water expressed as milligrams per litre of equivalent
calcium carbonate. In natural waters, these base are primarily
carbonate and bicarbonate ions. Another way to think of
alkalinity is in terms of basicity and resistance to pH change.
The amount of acid required to cause a specified change in pH in a
given volume of water increases as a function of the total alkalinity
levels of the waters. In general, early morning pH is greater
in waters with moderate or high total alkalinity than in waters
with low total alkalinity. The availability of carbon dioxide for
phytoplankton growth is related to alkalinity. Waters with total
alkalinities less than 15 or 20 mg/litre usually contain relatively
little available carbon dioxide. Waters with total alkalinities of
20 to ISO mg/litre contain suitable quantities of carbon dioxide
to permit plankton production for fish culture. Carbon dioxide
is often in low supply in waters with more than 200 to 250 mg/
litre of total alkalinity. The afternoon pH in waters with low
total alkalinity may often be as great as in waters with moderate
or high total alkalinity. Waters of low alkalinity are poorly
buffered against pH change and the removal of carbon dioxide
results in rapidly rising pH.

The total concentration of divalent metal ions (primarily

calcium and magnesium) expressed in milligrams per litre of
equivalent calcium carbonate is termed the total hardness of
water. Total alkalinity and total hardness values are normally
similiar in magnitude because calcium, magnesium, bicarbonate,
and carbonate ions in water are derived in equivalent quantities
from the solution of limestone in geological deposits. However,
in some waters total alkalinity may exceed total hardness and
vice versa. If total alkalinity is high and total hardness low, pH
may rise to extremely high levels during periods of rapid photo-
synthesis.
 f*
Desirable levels of total hardness and total alkalinity for ft*
culture generally fall within the range of 20 to 300 mg/Iitre.

If total alkalinity and total hardness are too low, they may be
raised by liming. However, there is generally no practical *way
of decreasing total alkalinity, and total hardness when they are
above the desirable level. As a general rule, the most productive
waters for fish culture have total hardness and total alkalinity
values of approximately the same magnitude. For example, a
water with a total alkalinity of ISO mg/litre and a total hardness
of 25 mg/litre is not as good for fish culture as a water in
which the total alkalinity is ISO mg/litre and the total hardness
is 135 mg/litre.

1.13. Aquatic weeds

Large aquatic plants (aquatic macrophytes) which may grow
in fish ponds are usually undesirable. They interfere with fish
management operations'such as seining, feeding and fish harvest,
compete with phytoplankton for nutrients, provide havens for
prey fish to escape predatory fish and thus encourage unbalanced
fish populations, favour mosquito production and contribute to
water loss through evapotranspiration. Aquatic macrophytes
includefilamentousalgae and submerged, floating-leafed, floating
and emergent macrophytes. Aquatic macrophytes which being
their growth at the pond bottom are limited to relatively trans-
parent waters. Therefore, management procedures which favour
plankton turbidity will often eliminate macrophytes(13).
Obviously, floating or floating-leafed macrophytes must be
controlled by other methods.

BIBLIOGRAPHY

ALMAZAN, G. AND C. E. BOYD 1978. An evaluation of Seochi disk visibility

for estimating plankton density in fish ponds. Hydrobiologia, 65 :601-608,

BARICA, J. 1975. Summerkill risk in prairie ponds and possibilities of its

prediction, / . Fttft. R&. Bd. Canada^ 32 (2834/ 288.
ts
BIASLEY, P. G. 1963. The penetration of light and die concentration of
dissolved oxygen in fertilized pond waters infested with Microcystis.
Proc. Ann. Conf. Southeastern Assoc. Game and Fish. Comm., 17 :222-226.

BOYD, C. E. 1976. Water chemistry and plankton in unfertilized ponds in

pastures and in woods. Trans. Amer. Fish. Soc., 105 :634-636.

• i i . 1982. Water quality management for pond fish culture. Else-

vier Sci. Publ. Co. Amsterdam. 318 pp.

- — , E. E. PRATHER AND R. W. PARKS 1975. Sulden mortality of a

massive phytoplankton bloom. Weed Set., 23 :61-67.

—•— , R. P. ROMAIRE AND E. JOHNSTON 1978. Predicting early morning

dissolved oxygen concentrations in channel catfish ponds. Trans. Amer.
Fish. Soc., 107:484-492.
HART, J. S. 1944. The circulation and respiratory tolerance of some Florida
freshwater fishes. Proc. Fla. Acad. Sci., 7:221-246.

PLUMB, J. A., J. M. GRIZZLE AND J. DEFIOUEIREDO 1978. Necrosis and

bacterial infection in channel catfish (Ictaluruspunctatus) following hypoxia.
/ . Wildlife Diseases, U: 247-253.

SCHROEDER, G. L. 1975. Nighttime material balance for oxygen in fish

ponds receiving organic wastes. Bamidgeh, 27:65-74.

SWINGLE, H. S. 1961. Relationship of pH of pond waters to their suitability

for pondfish culture. Proc. Pacific Sci. Congress 9 (1957), 10 (Fisheries):
72-75.
 FISH FEEDING AND WATER QUALITY
1
Fish eat most of the feed applied to ponds, but on a dry
matter basis, only a small percentage of the chemical substances
in feed are converted to fish flesh. To illustrate, in enamel
catfish culture, feed conversion values (weight feed applied/
weight fish produced) are about 1.5. Thus, if 3,000 kg of feed
were provided to fish in a 1-ba pond, 2,000 kg of fish could be
expected. However, the fish are about 25% dry matter and
the feed is about 92 % dry matter. Thus, 2,760 kg of dry matter
in feed might result in 500 kg of dry matter in fish — a dry
matter conversion ratio of 5.52. The difference between dry
matter in feed and in fish, 2,260 kg represents chemical sub-
stances that reach the water in metabolic wastes. The wastes
includes carbon dioxide, ammonia, phosphate and other organic
and inorganic substance.
Nutrients from metabolic wastes stimulate phytoplankton.
In a oatfish pond receiving upto 50 kg/ha of feed per day (1),
the metabolic waste from the production of 1 kg of live catfish
resulted in 2.69 kg of dry weight of organic matter id phyto-
plankton. Furthermore, fish excrete ammonia, each kg of live
catfish resulted in 0.061 kg of ammonia (lj. Thus, as feeding
rates increase, phytoplankton density and ammonia concentra-
tion increase in fish ponds. For exmple, at feeding rate of
50 kg/ha/day, chlorophyll a concentrations averaged 45 /ig/
litre in catfish ponds; at 100kg/ba/day of feed, chlorophyll
a averaged 75 /ig/litre. Of course, peak chlorophyll a coriofti-
trations were much greater —150 and 250 /*g/Utre respectively,
at feeding rates of 50 and 100 kg/ha/day. Total ammonia
nitrogen averaged 0.5 mg/litre at 50 kg/ba/day of feed and
0.9 mg/litre at 100 kg/ba/day. Of course, peak concentrations
were higher.

The density of phytoplankton during the course of a growiig

season ii also related to feedfeg rate. For example, as the
feeding ra(e was gradually iaoreased in a catfish f o w l ,
30
phytoplankton abundance.increased as evident from a decrease in
Secchi disc visibility (Fig. 2.1).
Dissolved Secchi disk
oxyflen, visibility,
mq/liter cm
10 100

Secchi disk 80
visibility

60

dissolved
oxygen
40

20

>L_
10 ' 20 30 40 50
J°
Feed, kg/ho per doy

Fro. 2.1. Secchi disc visibilities and early morning (0630 hrs.) dissolved
oxygen concentrations in a channel catfish pond as the feeding rate was
gradually increased from May to September.
'.,, As phytoplankton density increases with feeding rate, there
is a decrease in the early morning dissolved oxygen concentra-
tion (Fig. 2.1). In ponds stocked at different rates with channel
catfish (3), there was good agreement between feeding rates,
Secchi disc visibility (phytoplankton abundance) and early
morning dissolved oxygen concentrations (Table 2.1). If feeding
rate continues to increase, a level is reached where phytoplankton
abundance is so great that dissolved oxygen depletion may occur
each night. Without aeration, fish cannot survive at very high
feeding rates. Of course, even with aeration and adequate
dissolved oxygen concentrations, feeding rates may become so
high that high ammonia concentrations limit fish production.
 3i
As feeding rates increase, fish production increases. How-
ever, because ammonia concentration increase and dissolved
oxygen concentrations during the night decrease, conditions for
fish production gradually decline as feeding rates increase. For
example, at feeding rates of 34, 56, 78 and 92 kg/ha of feed
per day, feed conversion values for channel catfish in ponds
were 1.3, 1.7, 2.5 and 6.3 respectively. Hence, as feeding
rates andfishproduction increase in un-aerated ponds, the amount
of feed required to produce a given weight of fish increases.
Because of the diminishing returns from feed, a point is reached
where applying more feed becomes uneconomical. -

TABLE 2.1. Average early morning dissolved oxygen concentrations

and Secchi disc visibilities in channel Catfish ponds • „
with different feeding rates

Early
morning Secchi
Feeding rate dissolved disc
oxygen visibility
(mg/iitre) (cm)

Low (34 kg/ha/day) 4.71 68

Medium (56 kg/ha/day) 2.95 45
High (78 kg/ha/day) 1.95 2\

In aerated ponds, feed conversion values were essentially

constant (1.3 to 1.8) between feeding rate values of 25 and 125
kg/ha/day. At 150 kg/ha of feed, the feed conversion
value was 3.0. At 200 kg/ha of feed per day, the feed
conversion value was over 15. Dissolved oxygen concentrations
were adequate in all ponds and the increase in feed conversion
values at high feeding rates was attributed to high ammonia
concentrations.
The ultimate limit on feeding rate and fish production is
set by water quality. As a general rule,fishproduction increases
linearly with feeding rate while water quality deteriorates
exponentially with feeding rate.
Odours and flavours described as ' earthy-musty' are often
detected in fish from ponds. These odour and flavours originate
il
from organic substances that are syntheslsed and excreted into
the water by blue-green algae and aotinomycetes. The sub-
stances are absorbed by fish and impart an ' off-flavour 'tofish
flesh. Off-flavour is an important problem in the commercial
culture of fish, for off-flavour fish may not be suitable for
market. The environmental factors that favour off-flavour in
pond fish are not understood. However, the problem is worse
during warm weather and in ponds with high feeding rates (2).
Many techniques for preventing off-flavour in pond fish have
been tried, but none have been successful in all cases. At present*
the only known means of immediately removing off-flavour
from fish is to hold the fish in fresh water for several days.
This technique is usually impractical. Fortunately, off-flavour
problems usually disappears from pond within a few weeks to
a few months. Many times, through the necessity to hold
fish for a indeterminable period while off-flavour disappears is
an inconvenience and added cost tofishfarmers.

BIBLIOGRAPHY

BOYD, C. E. 1985. Chemical budgets for channel catfish ponds. Tram.

Amer.Fish. Soc, 114(in press).

BROWN. S. W. AND C. E. BOYD 1982. Off-flavor in channel catfish from

commercial ponds. Ibid., I l l : 379-383.

TUCKER, L., C. E. BOYD AND E. W. MCCOY 1979. Effects of feeding rate on

water quality, production of channel catfish and economic returns.
Ibid., 108; 389-396.
 4
LIMING t
it

4.1. Introduction f
Phytoplankton growth in waters with low alkalinity is often
limited by inadequate carbon dioxide and bicarbonate ion. Some
waters of low alkalinity are so acid that flsh do not survive
or grow well. Muds in ponds with low total alkalinity are
acid and strongly absorb the phosphorus added in fertilizer.
The addition of a liming material increases the pH of bottom
muds and makes phosphorus more available. Liming also
increases the availability of carbon for photosynthesis by rais-
ing the total alkalinity of the water. The greater total alkalinity
after liming results in a higher concentration of bicarbonate ion
which is in equilibrium with carbon dioxide. Liming also
increases the pH and total hardness of pond waters. Ponds
with total alkalinity values less than 10 mg/litre seldom produce
adequate plankton for good fish production unless they are
limed. Responses to fertilization are variable in unlimed ponds
with waters containing 10 to 20 mg/litre total alkalinity, but
unlimed ponds with waters above 20 mg/litre total alkalinity
consistently, produce adequate plankton after fertilization to
allow good fish production provided all other factors are favour-
able (5).
The decision to lime a pond should always be based upon
total alkalinity measurements rather than guesswork. Ponds in
same general area may differ greatly in total alkalinity. For
example, most ponds near Auburn, U.S.A. will benefit from
liming, but among these ponds are a few which have total alka-
linity values well above 20 mg/litre. The " rule of thumb "
recommendation that all ponds in the vicinity of Auburn,
Alabama need lime would result in unnecessary and wasteful
application of lime to some ponds. In determining whether
or not to lime a pond, one should recognize that there is no
single total alkalinity value below which lime is undoubtedly
needed. Experience has shown that liming is of little or no
46
bsnefit if total alkalinity is above 20 mg/litre. At total alkalinity
values telow 20 mg/litre, judgment must be used to decide
whether or not to lime because the need for lime increases with
decreasing total alkalinity. In ponds with total alkalinity values
between 15 and 20 mg/litre, the response to liming may be too
slight to justify the effort and expense. One should not use
lime in a pond if fertilization is not to be employed because
liming alone will not appreciably increasefishproduction except
in waters that are so acid thatfishwill not survive or grow at
normal rates. Furthermore, lime is seldom needed in ponds
where fish are supplied feed and do not depend upon naturally
occurring organisms for food.

4.2. Lime requirement

When lime is applied to a pond, it reacts with the mud and
until enough lime is added to satisfy the lime requirement of
the mud, little if any of the added lime will be available to
increase the pH, total alkalinity and total hardness. A lime
requirement procedure is available for determining the amount
of lime needed to raise the total alkalinity above 20 mg/litre
in ponds (2, 4). A simplified lime requirement technique
is provided in Chapter 7. This procedure is based upon a chemical
analysis of a mud sample and many fish culturists and biologists
will bi unable to obtain a lime requirement value for their pond.
In this event, liming material equivalent to about 2,000 kg/ha
of calcium carbonate may be applied and the total alkalinity
determined after 1 or 2 months. If the total alkalinity is still
too low, another application equal in amount to the first should
be made and the total alkalinity measured again. This proce-
dure should be repeated until enough lime has been applied to
maintain the total alkalinity above 20 mg/litre. The addition
of liming material equal to about 2,000 to 6,000 kg/ha of calcium
carbonate should suffice for most ponds. However, some ponds
which have high concentrations of organic matter in bottom
muds or have sulphide deposits in bottom muds or on their water-
sheds may require much greater amounts of lime. Occasionally
the lime application rate may be so high that cost of the lime
will be prohibitive and the water will be unsuited for fish
culture.
 47
Typical effects of liming may be illustrated by results of
experiments conducted at Auburn University (1). Agricultural
limestone (finely crushed dolomite) was applied to five ponds
at the rate of 4,000 kg/ha andfiveponds served as the unlimed
controls. All ten ponds were fertilized. Liming caused a
Mg/lifer " ""*"'<•'»,>• -
asCaCOs LIMED UNLIMED
50r o——o total hardness <
o o total alkalinity » •
P.

| F ' M ' A ' M ' J ' J ' A ' S

FIG. 4.1. Effect of application of agricultural limestone on total hardness

and total alkalinity in fish ponds.

marked increase in total hardness and total alkalinity (Fig.

4.1). The pH of bottom muds increased in the limed ponds
(Table 4.1). The production of Tilapia was 25% greater in the
limed ponds than in the control ponds. The total alkalinity of

TABLE 4.1. Average mud pH values for five limed and five unlimed
ponds — lime applied between February 17 and March 17, 1983

November August January

Type pond
1972 1973 1974

Limed 5.2 6.7 6.8

Unlimed 5.4 5.5 5.5
48
these ponds before liming averaged 13.5 mg/litre. Even greater
responses to liming have been reported in waters which had
lower total alkalinity before liming.

4.3. Applying lime to ponds

Agricultural limestone is the best liming material to use in
ponds. The material should be finely ground (particles should
pass through a sieve with 0.025 cm openings) and have a high
neutralizing value. Small particle size is necessary so that the
agricultural limestone will react quickly with water and mud.
The neutralizing values of liming materials refer to the amounts
of acid they will neutralize, expressed as a percentage of the
amount of acid neutralized by an equal amount of pure calcium
carbonate (3). A method for determining neutralizing value
is provided in Chapter 7. Thus, pure calcium carbonate
has a neutralizing value of 100% and is used as a standard
in referring to liming rates. In other words, if one wants to
apply a liming material at a rate equal to 2,000 kg/ha of calcium
carbonate and the liming material has a neutralizing value of
80%, he should apply 2,500 kg/ha (2,000 kg -?- 0*80) of the
liming material.

Hydrated lime [calcium hydroxide, Ca(OH)a] and burnt

lime (calcium oxide, CaO) have higher neutralizing values
than agricultural limestone, bat if applied in large quantities,
hydrated lime and burnt lime cause excessively high
pH and fish mortality. Hydrated lime and burnt lime are
sometimes applied to Waters which contain no fish or to muds
of ponds which have been drained to raise the pH and kill fish
disease organisms. Basic slag has been used as a liming material
in fish culture, but its neutralizing value is lower than that of
most agricultural limestones and extremely large applications of
basic slag are required. Agricultural gypsum (calcium sulphate,
CaS04-2HaO) is not a liming material, although it has incor-
rectly been used as one by some fish culturists.
Liming materials can be easily broadcast over the bottoms
of empty ponds, but application is more difficult when ponds
are full of water. Best results may be achieved by broad-
casting the liming material over the entire pond surface. Bags
 40
of liming material may be emptied from a moving boat. Bulk liming
material is cheaper and may be applied from a plywood platform
attached between two boats. Liming materials should be applied
during the late fall or early winter in temperate regions so that
it will react with waters and muds before fertilizers are applied
in the spring. In tropical regions, lime should be applied at
least 1 month before fertilizer applications are initiated. This
is important because liming materials will precipitate phosphorus
if applied at or near the same time as fertilizers. However, once
the liming material has reacted with the mud, greater availability
of phosphorus fertilizer will result. The residual effect of
liming is governed by the rate of water loss to seepage and pond
overflow. In ponds with normal rates of water loss, liming will
usually last for 3 to 5 years. Once a pond has been limed,
small annual applications (20% to 25 % of the initial application
rate) may be used to avoid having to make large applications
of lime every few years.

BIBLIOGRAPHY

ARCE, R. G. AND C. E. BOYD 1975. Effects of agricultural limestone on water

chemistry, phytoplankton productivity and fish production in soft water
ponds. Trans. Amer. Fish. Sac., 104: 308-312.

BOYD, C. E. 1982. Water quality management for pond fish culture. Elsevier
Sci. Publ. Co. Amsterdam. 318 pp.

1974. Lime requirements of Alabama fish ponds. Auburn Univ.

{Ala.) Agr. Exp. Sta., Bull., 459: 20 pp.

PIIXAI, V. K. AND C. E. BOYD 1984. A simple method for calculating liming

rates for fish ponds. Manuscript submitted to Aquaculture.
THOMASTON, W. W. AND H. O. ZBIAER 1961. Results of a six-year investi-
gation of chemical soil and water analysis and lime treatment in Georgia
Fish Ponds. Proc. Am. Conf. Southeastern Assoc. Game and Fish Comm.,
15:236-245.

4
5
DISSOLVED OXYGEN AND AERATION

5.1. Introduction
Almost all problems with dissolved oxygen in fish culture are
the consequences of heavy plankton blooms. In fertilized ponds,
fertilization should be halted when plankton blooms get too
dense (i.e., Secchi disc readings of 25 cm or less). In ponds
where fish are supplied feeds, heavy plankton blooms are the
natural consequences of high feeding rates. Lower feeding rates
will result in less plankton growth, but only at the expanse of
decreased fish production. Suitable plankton densities result in
Secchi disc visibilities of 30 to 60 cm. The probability of
problems with low dissolved oxygen concentration increases as
the magnitude of the Secchi disc visibility decreases below 30 cm.
In ponds with Secchi disc visibility values of 10 to 20 cm,
dissolved oxygen concentrations may fall so low at night that
fish are stressed and a cloudy day may lead to dissolved oxygen
depletion before the next morning. Aeration is an effective
means of preventing fish mortality when dissolved oxygen is low
and aeration can bs used to permit high levels of fish
production. ........

5.2. Management of dissolved oxygen

A number of procedures are used to prevent fish kills when
dissolved oxygen concentrations are dangerously low. Application
of upto 6 or 8 mg/iitre of potassium permanganate has been
frequently recommended, in the United States. The potassium
permanganate is supposed to oxidize organic matter and lower
the demand for dissolved oxygen in the pond. Recent research
at Auburn University has demonstrated that potassium perman-
ganate is entirely worthless for this purpose and that its
application actually increases the length of time required for
dissolved oxygen concentrations to return to normal levels.
Applications of calcium hydroxide have been recommended to
 51
destroy organic matter in ponds with low dissolved oxygen
concentrations and thereby reduce rates of oxygen consumption
by bacteria. There is no reason to believe that applications of
calcium hydroxide will lower concentrations of organic matter.
However, when dissolved oxygen is low, carbon dioxide is
usually quite high. The application of calcium hydroxide will
remove carbon dioxide which will allow fish to better utilize the
low concentration of dissolved oxygen. Each milligram per litre
of carbon dioxide will require 0-84 mg/litre of calcium hydroxide
for its removal. For example, if a pond contains 25 mg/litre of
carbon dioxide, a calcium hydroxide treatment of 21 mg/litre
(25 mg/litre x 0.84) would remove the carbon dioxide. Following
phytoplankton die-offs, applications of fertilizers have been
employed to encourage phytoplankton growth and foster dissolved
oxygen production. Research to document the effectiveness of
this procedure has not been conducted, but nutrient concentrations
are already high in ponds following phytoplankton die-offs and
it is doubtful that fertilization is necessary.

The only really effective procedure for/-preventing fish

mortality during periods of extremely low dissolved oxygen
involves the use of mechanical devices. Large, tractor-powered
pumps may be used to pump fresh, oxygenated water from a
nearby pond into the pond with low dissolved oxygen -concentra-
tions. Alternatively, water from wells, spring, streams, etc.-
may be released into the oxygen depleted pond. Well water
should b ; discharged across a baffle for aeration because well
water is often high in carbon dioxide and deficient in dissolved
oxygen. When oxygenated water is released into a pond with
oxygen depletion, bottom water which contains less dissolved
oxygen and more carbon dioxide than surface water should
simultaneously be released from the pond if possible. Pumps
may also be used to pump water from the pond with low
dissolved oxygen and release this water, with.force back onto the
surface of the same pond. The agitation and circulation of the
water increases its dissolved oxygen content. However, this
method is not as effective as pumping fresh, oxygenated water
from another pond or well into the pond with low dissolved
oxygen. Various types of aeration devices may be used to
introduce oxygen into waters with low dissolved oxygen
52
concentrations. Small spray-type surface aerators are in common
use. These aerators are most effective in small ponds or when
several are operated in a large pond. More powerful aerators
such as the CrisafuUi pump and sprayer and the paddle-wheel
aerator supply considerably more dissolved oxygen to ponds
than the spray-type surface aerators. However, CrisafuUi pumps
and paddle-wheel aerators are expensive and must be operated
from the power take-off of a farm tractor. Large fish farms and
research stations can afford to maintain and operate emergency
aeration equipment, but small scale fish culturists have little
recourse when faced with dissolved oxygen problems. Fortu-
nately, problems with dissolved oxygen seldom occur except in
ponds where fish are fed at high rates.

Dissolved
oxygen,
rhg/liter

9- measured
volues

\1
8-
7-
61-
5
h
4
3 projected
value
2
I
0
8p.m. 18 p.m.
% \

4o.m.
i
FIG. 5.1. A graphical method for predicting the night time decline in
dissolved oxygen concentration infishponds.

Fish culturists often monitor dissolved oxygen concentrations

during the night in ponds to determine if emergency aeration is
needed. Recent research has resulted in procedures for predicting
how low dissolved oxygen concentrations will fall during
 33
the night. Such predictions permit the fish culturist to prepare
for emergency aeration in advance. The simplest of these
procedures involves the measurement of dissolved oxygen
concentrations at dusk and two or three hours later. These two
values are plotted versus time oh a graph and a straight line is
projected through the two points and used to estimate the
dissolved oxygen concentration at later hours during the night.
The use of this technique is illustrated in Fig. 5.1.

5.3. Aeration and fish production

Continuous or night time aeration can increase production of
catfish in ponds, as results of four studies summarized below
clearly demonstrate.

Loyacano (5) aerated white catfish ponds with a blower that

forced air through openings in pipes on pond bottoms. Because
of improved water quality, higher feeding rates were employed in
aerated ponds. Three ponds aerated at 10.5 m* of air /minute/ha
had average net fish production of 5,500 kg/ha. Average net
flsh production for three unaerated ponds was 2,700 kg/ha.

Parker (7) used air-lift pumps for aeration and obtained a

maximum harvest weight of 15,800 kg of channel catfish/ha and
unaereted ponds yielded 3,000 kg/ha. Of course, stocking and
feeding rates were much higher in aerated ponds. In spite of
aeration with air-lift pumps, low dissolved oxygen (DO) in some
ponds necessitated additional aeration with splasher-type surface
aerators. Parker also flushed one volume of water through
ponds every two weeks.

Plemmons (8) employed continuous aeration with splasher-

type surface aerators at 5.5 kw/ha. Unaerated ponds stocked at
12,200 channel catfish/ha had an average harvest weight of
2,500 kg/ha. Aerated ponds stocked at 30,100 fish/ha yielded
12,800 kg of catfish/ha. Plemmons occasionally employed
emergency aeration with a Crisafulli pump to prevent DO
depletion even though ponds were continuously aerated with small
surface aerators. He also flushed appreciable water through
some ponds in hopes of improving Water quality.
54
Hollerman and Boyd (4) (stocked channel catfish at 19,770/ha
in 12 ponds. Six ponds were aerated for two to six hours per
night with splasher-type aerators at 5.5 kw/ha and six ponds
were not aerated. Emergency aeration and water exchange
Were not employed. Fish kills were common in unaerated ponds,
but rare in aerated ponds. Harvest weights averaged 1,400 kg/ha
in unaerated ponds and 5,390 kg/ha in aerated ponds.

In all four cases summarised above, the increase in fish

production was attributed to higher concentrations of DO
resulting from aeration. The studies also demonstrated that
aeration will not always prevent DO depletion. Stocking, feeding
rates and oxygen demands were higher in aerated ponds. At times,
demands for oxygen exceeded abilities of small aerators to supply
oxygen and emergency aeration with larger aerators and water
exchange was employed to prevent fish kills. Economic analyses
of data collected by Parker (7), Plemmons (8) and Hollerman
and Boyd (4) suggested that aeration could increase profits.
However, it is difficult to extrapolate economic data from small
research ponds to actual fish-farm conditions.

Benefits of aeration have been clearly illustrated in research.

However, because of a lack of understanding of the principles of
aeration and of aerator function, fish farmers often fail to achieve
the maximum benefits of aeration. Therefore, some basic facts
are provided bilow about aerators and aeration.

5-4. Oxygen transfer

At a specified combination of water temperature, salinity and

atmospheric pressure, water contains a given concentration of
oxygen at equilibrium (saturation). For freshwater fish ponds in
the southeastern United States, temperature is the major factor
causing differences in DO concentrations in saturated waters. *,
If the DO concentration is below saturation, oxygen will diffuse
from the air into the water until saturation is finally attained.
The driving force causing net movement of oxygen from air to
water is the oxygen saturation deficit (DO at saturation minus
actual DO concentration). The greater the saturation deficit, *
the faster oxygen will enter, the water. f
 £f
• ' Oxygen must enter the water through the surface film, but if
the DO concsntration in a body of water is to rise, oxygen
entering the film must be mixed ^throughout the water column.
Water in the film quickly increases in DO concentration, but
unless the oxygen-enriched water in the film is mixed with
underlying water of lower DO concentration, the rate of diffusion
of oxygen into the surface film will be greatly retarded. The
movement of oxygen through still water is slow, so turbulence is
necessary to maintain a large oxygen saturation deficit between
surfacefilmand air. Turbulence (mixing) continuously replaces
oxygen-enriched water in the surface film with water of lower
DO concentration.
The rate of diffusion of oxygen into water depends primarily
upon the oxygen saturation deficit, the ratio of water surface to
Water volume and the degree of turbulence. In a given body of
water, natural aeration is largely a function of the saturation
deficit and wind velocity. Waves increase the surface to volume
ratio and turbulence mixes surface water with underlying water.
Aerators also provide oxygenation by increasing the area of
contact between air and water and the amount of turbulence.

5-5. Types of aerators

Aerators increase the area of contact between air and water
by (a) agitating surface water, (b) releasing air bubbles beneath
the surface or (c) both. While oxygenating water, aerators also
impart energy to water and cause horizontal and vertical mixing.
Flexibility in aerator design permits many types of aerators, a
few of which are described below.
Diffused-air aerators employ an air blower or air compressor
and porous pipe to release air bubbles at the pond bottom (Fig.
5.2). The efficiency of oxygen transfer is related to bubble size —
small. bubbles offer a greater air-water interface than do large
bubbles. Water depth also influences efficiency, because the
deeper the bubbles are released, the greater the contact time
between bubbles and water.
Venturi aerator suck air into water so that bubbles are formed
(Fig. 5.3). The aspirating-propeller-pump aerator is a modifica-
56
tion of the venturi aerator in which a propeller at the end of a
hollow shaft imparts velocity to water and sucks air down the
shaft. This air is mixed into the turbulent water asfinebubbles.

SLOWER

POROUS PIPE'
Fio. 5.2. Diffased-air aeration.

AIR

XixWS.':/::

^PUMP
FIG. 5.3. Venturi aerator.

U-tube aerators (10) are usually 12 to 18 m deep (Fig. 5.4),

hence bubbles have a long contact time with water. Unless
adequate head is available, water must be pumped through the
U-tube.
f
An air-lift pump (Fig. 5.5) consists of an open-ended pipe or
tube into which air is released. The air bubbles rise through therf
tube and effect oxygenation. Of course, the movement of bubbles
through the pipe results in pumping of water.
 «

MR BLOWER

V* Kj
Q
• » , -

\o,
FlQ. 5,4. U-tylje aerjto?.

WATER ANO
AH«
'All*

— - •• "• """ *~J^C"-~ '** '

' ""•" • • —
" — " ^ ^ ^ — •

9 tr"'' ^' ^ **

i
• - II - ""
~ ^-L^*» * "" ll —T"
*
B,UBBLES^~ -
•
j * — II " ""—
— _ * • * * * * " " *

Fio. 5.S. Airlift pump aerator.

58
Splasher-type surface aerators (vertical turbines) jet water
into the air (Fig. 5.6). Water is broken into turbulent, thin
layers and drops that have large surface-to-volume ratios.
Paddle-wheel aerators are another common type of surface
aerator.

SPLASH CONE
/ AND WEIGHT

\ /

y^K L
FLOAT

AgSZ
/ \
/
roR
SUBMERSED MOTOI
AND PROPELLER
FIG. 5.6. Splasher-type surface aerator.

Gravity aerators employ head loss. Water falls over a weir

or from a pipe onto a splash board (Fig. 5.7), paddle-wheel,
brush or inclined plane. Water may be pumped to the top of a
vertical pipe and allowed to fall through perforated aprons.

FIG. 5.7. Gravity-flow aeration using a splash board.

 59
Chesness and Stephens (2) demonstrated that all gravity aerators
mentioned above were effective in oxygenating water.
Colt and Tchobanoglous (3) presented standard oxygen
transfer rates for various types of aerator (Table 5.1). With the
exception of the highly efficient U-tube areator, all aerator had
oxygen transfer rates of 0 6 to 2*4 kg oxygen/kw/hour. The
U-tube aerator is highly efficient, but is difficult and expensive to
construct and has virtually no application in pondfishculture.

TABLE 5.1. Typical rates of oxygen transfer under standard conditions for
aeration systems used in fish culture (3)

Transfer rate
Aeration system (Kg oxygen/Kw/
hour)
Diffused-air systems
Fine bubble 1.2—2.0
Medium bubble 1.0—1.58
Coarse bubble 0.6—1.2
Low-speed surface aeration (with or without draft tube) 1.2—2.4
High-speedfloatingaerator 1.2—2.4
U-tube aerator 4.5-45.6
Gravity aerator 1.2—1.8
Ventuxi aerator 1.2—2.4
Static tube systems 1.2—1.6

5-6. Testing aerators

Aerators are tested in small basins — often 200,000 litres or
less. A basin isfilledwith clean tap water and DO is removed
with sodium sulphite and oobalt chloride. Sodium sulphite
reacts with oxygen as follows :
Na,SO, + 1/2 O, = Na8SOt
Theoretically, 7.9 mg/iitre of sodium sulphite are needed to
remove 1 mg/litre of DO. However, because the aerator is used
to mix the sodium sulphite, some oxidation of sulphite occurs
during the mixing period. About 1.5 times the theoretical
quantity of sodium sulphite normally is added. Cobalt chloride
serves as a catalyst — it is applied at 0*05 to 01 mg/litre of cobalt.
Once the water is deoxygenated, the aerator is operated to raise
60
the oxygen concentration in the basin. Measurements of DO are
made at intervals (a minimum of six to ten times while the DO
rises from 0 to 70 or 80 % of saturation) at different locations (at
least four to six locations) in the basin. The plot of the natural
logarithm of the saturation deficit versus time should give a
straight line, the slope of which is the oxygen transfer coefficient
(KLa). The KLa is calculated by the equation
K* In (Cs - Q ) - In ( C - Cg)
KLa - -——
Tt — l j
where
KLa = oxygen transfer coefficient in hr- 1 ;
Cs = DO concentration at saturation in mg/litre ;
Cj <= initial DO concentration is mg/litre;
C8 s= final DO concentration in mg/litre;
t, = time that DO concentration reaches 10 or 20% of
saturation (Cx) in mg/litre;
ta sa time that DO concentration reaches 70 or 80% of
saturation (Ca) in mg/litre.
The KLa calculated by the above equation is for the temperature
of water in the test basin. By convention, KLa values are
corrected to 20°C [ (K^a),,,]:
(KLa)T = (KLa)80 x 1024T-*°
where
T = temperature of water in the test basin in °C.

The amount oxygen transferred per hour at standard

conditions (tap water; 20°C; 0 mg/litre DO) may be obtained by
(OT)M *, (Kta)26 X C* x V x 10-«
where
(OT)ao = oxygen transfer in kg/hr under standard conditions
(tap water, 20°C, 0 mg/litre DO);
C»„ = DO concentration at saturation and 20°C in mg/litre;
V = Volume of test basin in litres.

Additional information on aerator tests are presented by the

American Public Health Association et al (1) and Stuckenberg
etal (11).
 61

5.7. Oxygen transfer in ponds

Pond conditions are different from those in test basins and
oxygen-transfer rates for ponds are less than those obtained in
test basins and reported by manufactures. Nevertheless, oxygen
transfer ratings are useful in comparing the capabilities of
different aerators.

If pond waters are saturated with oxygen, aerators cannot

transfer more oxygen. However, the further below saturation
the DO, the more efficiently aerators transfer oxygen and the
greatest efficiency is for 0 mg/litre DO. Aerators are rated for
20°C and pond waters are usually warmer. The rate of
oxygenation of tap water if usually greater than that of pond
water and tap water often holds more DO at saturation than
does pond water of the same temperature. The following
equations are used in obtaining factors for relating aeration of
tap water to aeration of pond water (12):
.-,•'.. (Kt,a)ift Pond water
(KLa)M Tap water
o CJo Pond water
* "* C w Tap water
Shelton and Boyd (9) determined <£ and £ values for 43 samples
of pond water. The 0 factors ranged from 0-92 to 1*00 and the
mean standard deviation was 0-98 ± 00-19. Values for 84% of
the ponds were between 0-96 and 1-00. The o( factors for the 43
samples ranged from 0-66 to 1-07 with a mean of 0-94 and a
standard deviation of db 0-084. Most of the pond waters had
factors between 0-90 and 1'00. Alpha factors greater than 1-0
are not uncommon because waters may contain natural surfactants
that enhance oxygen transfer. Most factors for fish ponds were
somewhat larger than those for wastewater.

The following equation (6) corrects standard oxygen-transfer

rates for pond conditions:

(OT)p _ OTM [ ^ % ^ - (1.0MF-* ]

61
where
(0T) P = oxygen transfer rate under pond conditions in kg
0,/kw/hr;
(OT)2o = manufacturer's rating in kg Os/kw/hr;
Cp = DO Concentration in pond in mg/litre.

BIBLIOGRAPHY

AMERICAN PUBLIC HEALTH ASSOCIATION, AMERICAN WATER WORKS ASSOCIA.

TION, WATER POLLUTION CONTROL FEDERATION 1980. Standard methods
for the examination of water and wastewater. American Public Health
Assoc, Washington, D.C.

CHESNESS, J. L. AND J. L. STEPHENS 1971. A model study of gravity flow aerators

for catfish raceway systems. Trans. Amer. Soc Agric. Eng.,14 :1167-1169.

COLT, J. AND G. TCHOBANOGLOUS 1979. Design of aeration systems for aqua-

culture. Department of Civil Engineering, University of Calif., Davis.

HOLLERMAN, W. D. AND C. E. BOYD 1980. Nightly aeration to increase pro-

duction of channel catfish. Trans. Amer. Fish. Soc, 109 : 446-452.

LOYACANO, H. A. 1974. Effects xrf aeration in earthen ponds on water quality

and production of white catfish. Aquaculture, 3 : 261-271.

METCALF AND EDDY (Ed.) 1979. Wastewater Engineering ; Treatment, Disposal,

Reuse. McGraw-Hill, New York.
PARKER, N; C. 1979. Channel catfish production in continuously aerated
ponds. Research Workshop Summary of Papers, Catfish Farmers of
America Annual Meeting, Jackson, Mississippi.

PLEMMONS, B. P. 1980. Effects of aeration and high stocking density on

channel Catfish production. M.S. Thesis, Louisiana State Univ., Baton
Rouge, tousisnana.
SHELTON, J. L. AND C. E. BOYD 1982. Correction factors for calculating oxygen-
transfer rates for pond aerators. Trans. Amer. Fish. Soc, 112 : 120-122.

SPEECE, R. E. 1969. U-tube oxygenation for economical stratification of

fish hatchery water. Ibid., 89 : 789-795.

STUCKENBERO, J. R., V. N. WAHBEH AND R. E. MCKINNEY 1977, Experiences

in evaluation and specifying aeration equipment. / . Water Pollution
Control Fed., 49 : 66-82.

WHEATON, F. W. 1979. Aquacultural Engineering. Wiley-Interscience, New

York.
 6
MISCELLANEOUS TREATMENTS AND
CALCULATION OF TREATMENT RATES

6-1. Introduction
Three miscellaneous treatments for improving water quality in
ponds are covered in this chapter. These include turbidity
removal, aquatic plant control and pH reduction. Information
for calculating treatment rates is also provided.

6-2. Removal of cfay turbidity

In some ponds, it is necessary to remove the turbidity caused .
by suspended clay particles so that light will penetrate deep
enough into the pond for phytoplankton growth. The oldest
technique for removing clay turbidity involves the application of
organic matter (3, 5). Recommendations vary, but the most
popular include: two or three .applications of 2,000 kg/ha of
barnyard manure; one or more applications of 2,000 to
4,000 kg/ha of hay; and 75 kg of cottonseed meal phis 25 kg of
superphosphate per hectare at 2 to 3 week intervals. The
effectiveness of organic matter applications in removing clay
turbidity varies and several weeks must usually pass before one
can determine if a particular treatment was a success.
A better method for removal of clay turbidity is treatment
with filter alum (aluminium sulphate, A12(S04), • 14H,0).
Alum will cause suspended clay particles to coagulate and
precipitate from the water within a few hours (1). The exact
application rate for. alum may be determined by treating samples
of pond water in beakers with concentration of alum ranging
from 10 to 40 nig/litre at 5 mg/litre cdncentritions. intervals.
The lowest concentration of alum which clauses a floe of clay
particles to form within 1. hour is taken as the desired treatment
rate.. Many fish culturist will be unable to conduct this test, but
an application of 25 to 30 nig/litr? .will a^pjaWntry precipitate the
clay turbidity from" most ponds." When applying alum it should
64
bs dissolved in water and quickly distributed, preferably by
spraying, over the entire pond surface. Application should be
made during calm, dry weather because mixing by wind and
rain will break up the floe and prevent it from settling out.
The results of alum treatment in four ponds are illustrated in
Table 61.

TABLE 6.1. Effects of alum (Aluminium Sulphate) treatment on clay

turbidity infishponds

Alum applied Turbidity units Reduction in

Pond (rag/litre) - - turbidity (%)
Before After
treatment treatment

E-67 15 40 2 95
E-68 20 28 3 89
E-73 20 19 3 84
S-27 20 830 24 97

Alum has an acid reaction in water, so it destroys total

alkalinity and reduces pH. Each milligram per liter of alum will
decrease the total alkalinity by 0-5 mg/liter. If the total
alkalinity of water is below 20 mg/litre, alum treatment may
depress the pH to the point that fish are adversely affected.
Hydrated lime (calcium hydroxide, Ca(OH),) applied
simultaneously at the rate of 0-40 mg/litre for each 1-0 mg/litre
increment of alum will prevent unfavourable changes in alkalinity
and pH. Another alternative is to lime ponds that have waters
of low alkalinity before treating with alum. Liming materials
will often precipitate day turbidity, but if turbidity persists
after liming, alum treatment may bs conducted without danger
of pH depression.
Although alum treatment will clear pond water of clay
turbidity, it does nothing to correct the cause of turbidity.
Unless the source of the turbidity is eliminated, ponds will again
become turbid with clay particles. Clay turbidity usually results
because ponds receive large volumes of turbid runoff after each -
rain. Erosion of the watershed may be prevented by revegetation.
If this is imposible, it is sometimes possible to divert the turbid
runoff from the pond by use of a diversion ditch.
 65

6*3. Reduction of pH
Waters which have high total alkalinities and low total
hardnesses may have dangerously high pH values during periods
of rapid phytoplankton growth. Waters of this type do not
occur often, but one should analyse the water to determine if the
potential for high pH exists.

Liming to increase total hardness is of no value in preventing

high pH because lime application increases total alkalinity and
total hardness by roughly the same amount. Applications of
ammonium fertilizers have been recommended to lower the pH of
pond water. The ammonium ion in fertilizer is nitrified to
nitrate with the release of hydrogen ion which lowers the pH.
However, at high pH a large percentage of the ammonium ion
applied to a pond will immediately be transformed to un-ionized
ammonia which is highly toxic to fish. Filter alum (aluminium
sulphate) may be added to ponds to decrease pH. An alum
treatment equal in milligrams per litre to the phenolphthaelin
alkalinity will reduce the pH to approximately 8-5. Although
alum treatment may be used to prevent a fish kill when the pH is
too high, it does nothing to change the conditions responsible for
high pH.

Agricultural gypsum (oalcium sulphate) may be applied to

water to increase the total hardness without affecting the total
alkalinity. Experience indicates that gypsum will alleviate the
conditions responsible for high pH, but confirmatory research
is needed. The best treatment rate for agricultural gypsum
appears to be the amount which will increase the total hardness
to a level where it equals the total alkalinity. The treatment
rate may be determined from the following equation :
Agricultural
gypsum (mg/litre)=(Total alkalinity—Total hardness) x2-2

The agricultural gypsum should be applied in the same

manner as liming materials. The residual effect of gypsum
treatment is not known.
5
66

6-4. Aquatic plant control

As mentioned earlier, one effective technique of controlling

many species of macrophytes is through fertilization to produce
plankton turbidity and shade the pond bottom. This technique
is especially powerful if ponds are constructed so that no areas
are shallower than about 60 cm. Grass carp (white amur) eat
tremendous quantities of aquatic vegetation and provide a
biological method for controlling macrophytes. When stocked at
60 to 80 pir hectare, grass carp will control most species of
macrophytes that cannot be controlled by plankton turbidity.
Grass carp are even effeceive in controlling macrophytes in ponds
that are not turbid with plankton. In small ponds, macrophytes
may be controlled by cutting or by dragging them out with a rake
or seine.
Herbicides are also used in fish culture to control macrophytes.
The manufacturer's label gives the rate and method of application
for a herbicide. The label provides information on safety
precautions. Usually the concentrations of aquatic herbicides
used to kill macrophytes are safe to fish. Decay of macrophytes
killed by herbicides can cause dissolved oxygen depletion. If
ponds have extensive areas of macrophytes, 1/4 to 1/5 portions of
the pond should be treated at 1 to 2 week intervals to reduce the
chance of dissolved oxygen depletion. The major limitation of
herbicides for controlling macrophytes is that once the
concentration of a herbicide declines to a non-toxic level,
macrophytes will regrow. Thus, repeated applications of
herbicides are required to control macrophytes, often at
considerable expense.
Algicides are sometimes used eo control phytoplankton in
ponds. Copper sulphate, the most widely used algicide, will kill
most species of phytoplankton at concentrations of 0.1 to 0'5 mg/
litre in waters with total alkalinities below 40 to 50 mg/litre (6).
In waters with higher alkalinities, copper sulphate concentrations
of 1.0 mg/litre or more may be required to kill phytoplankton.
Copper sulhpate may be applied by dissolving it in water and
distributing it over the pond surface. Alternatively, copper
sulphate crystals may be placed in a burlap bag and the bag
 67
towed behind about until the copper sulphate dissolves. Burlap
bags of copper sulphate may be positioned in ponds so that the
chemical gradually dissolves and mixes with the water (2).
Copper sulphate may also be used to treat scums of phytoplankton
which drift to the leeward sides of ponds (4).
Phytoplankton killed by copper sulphate decomposes rapidly
and may result in low dissolved oxygen concentrations. Copper
sulphate has no appreciable residual toxicity and phytoplankton
growth will resume soon after treatment. Fish are susceptible to
copper sulphate and in waters with alkalinities less than 20
mg/litre, treatment with 0.5 to 1.0 mg/litre of copper sulphate
may kill fish.
Synthetic algicides such as Diuron (3-(3, 4-dichlorophenyl)-ll
1-dimethyl urea) and Simazine (2-chloro-4, 6-bis (ethylamino),
s-triazine) are sometimes used to kill phytoplankton. These
algicides are extremely toxic to algae, have a long residual
action and are not toxic to fish at concentrations used to kill
phytoplankton. As with copper sulphate, extensive mortality of
phytoplankton following applications of synthetic algicides may
result in depletion of dissolved oxygen. Some fish culturists
have attempted to ' thin' phytoplankton blooms by small,
periodic applications of synthetic algicides to ponds receiving
heavy applications of feed. However, recent research (7)
demonstrated that this practice results in prolonged periods of
low dissolved oxygen concentrations and reducedfishyields.

6.5. Calculating treatment rates

Concentrations for chemical treatments of ponds are given in
milligrams per litre sofishculturists must calculate how much of
a chemical to add to a pond to give the desired concentration.
To calculate the amount of a chemical needed, the volume of the
pond must be known. Assuming the surface area of a pond is
known, the simplest technique for obtaining the average depth to
use in computing the volume is to make transects (8 or 10 will
usually suffice) across the pond in a boat while making depth
soundings at regular intervals with a calibrated rod or
sounding line. The average of all soundings is taken as the
average depth.
68
Once the volume of a pond is knowu, it is a simple matter to
calculate treatment rates. Since 1 cubic metre (m3) weighs 1,000
kg and 1 gram contains 1,000 mg, then 1 gram per 1 m8 equals
1 mg/litre. The following examples illustrate how to calculate
amounts of chemicals to add to ponds.

Example : A pond has a surface area of 0.26 ha and an

average depth of 1.15 m. How much filter alum (100% pure)
must be applied to the pond to give an alum concentration of
25 mg/litre ?
(1) Since 0.26 ha = 2,600 ma, the pond volume is
2,600 m2 x 1.15 m = 2,2990 m3.
(2) Each cubic metre will require 25 g of alum for a concen-
tration of 25 mg/litre, so the amount of alum needed for the
entire pond is
2,990 m s x 25 g/m» = 74,750 g.
(3) A treatment of 74,750 g equals 74.75 kg.

Example : The average depth of a pond is 0.57 m and the

surface area is 0.01 ha. How much agricultural gypsum (8%
pure) must be applied to produce a gypsum concentration of
50 mg/litre ?
(1) Since 0.01 ha = 100 m2, the pond volume is
100 ma x 0-57 m - 57 m a .
(2) Each cubic metre will require 50 g of gypsum for a concen-
tration of 50 mg/iltre, but the agricultural gypsum is only 80%
pure. Therefore, we may calculate the concentration of
gypsum as follows:
50 g + 0-80«- 62.5 g.
The amount of agricultural gypsum needed for the entire pond
Will be
57 m3 x 62.5 g/m3 - 3,562 g or 3.56 kg.
Example : A pond with a volume of 1,000 m3 must be treated
with a herbicide. The herbicide is a liquid with 75% active
ingredient and a density of 0.85 g/ml (0.85 kg/litre). How much
of the liquid herbicide must be applied to the pond to give a
concentration of 1 mg/litre of active ingredient ?
 69
(1) The amount of the active ingredient to give a concentra-
tion of 1 mg/litre is
1000 m» x 1 g/m» x 1000 g - 1.0 kg.
(2) the herbicide has an active ingredient content of 75%, so
the weight of herbicide containing 1.0 kg active ingredient is
1.0 kg + 0.75 = 1.33 kg.
(3) The density of the herbicide is 0.85 kg/litre, so the volume
of the herbicide weighing 1.33 kg is
1.33 kg -5- 0.85 kg/litre - 1.56 litres.
Thus, 1.56 litres of the liquid herbicide would give a concen-
tration of 1 mg/litre of the active ingredient when applied to
the pond.

6.6. Applying chemicals to pond

Chemicals which are applied to ponds come in a variety of
formulation includings crystals, solutions, soluble powders,
emulsifiable concentrates and granules. Fish culture stations
and large fish farms can afford rather eloborate equipment for
applying chemicals. For example, chemicals may be dissolved
in a tank of water or some other solvent and sprayed over
the surface with a power sprayer. Liquids may be dispersed
uniformly from a boat mounted tank through a boom consisting
of a pipe with a series of small diameter holes in its underside.
A valve regulates the rate at which the solution is fed by gravity
into the water or a pump may be used to effect more uniform and
forceful release. Dispensers for granules or powder may consist
of hoppers with adjustable dispensing holes in the bottom. An
auger is employed to prevent clogging of holes. Finally,
chemicals may be released into the wash of an outboard motor
propeller to effect mixing as the boat moves over the pond
surface.

in instances where the owner of one or a few ponds must

apply chemicals, it is usually not practical to purchase or
construct an elaborate dispenser. The chemical can ba dissolved
or mixed in a large container of water and applied to the pond
surface. Application may be accomplished with a pressurized '
70
garden sprayer. However, if sprayer is not available the solu-
tion or mixture may be splashed with dipper over the pond
surface. Care should be taken to dispense the chemical as
uniformly as possible. Granules may be broadcast by hand or
with a small ' cyclone' seeder. Crystals may be placed in a
burlap bag and the bag towed behind a boat until they have
dissolved. Only a little ingenuity is needed lo develop a
method for applying a chemical to a pond once the treatment
rate has been established.

BIBLIOGRAPHY

BOYD, C. E. 1979. Aluminium sulfate for precipitating clay turbidity

from fish ponds. Trans. Amer. Fish. Soc, 108.

CRANCE, J. H. 1963. The effects of copper sulfate on Microcystis and zoo-

plankton in ponds. Prog. Fish-Cult., 25 : 198-202.
IRWIN, H. W. AND J. H. STEVENSON 1951. Physiochemical nature of clay
turbidity with special reference to clarification and productivity of im-
pounded waters. Okla. Agr. Mech. Coll. Bull., Arts and Sci. Studies,
Biol. Ser., 48:1-54.

KESSLER, S. 1960. Eradication of blue-green algae with copper sulfate.

Bamidgeh, 12 :17-19.

SWINGLE, H. S. AND E. V. SMITH 1947. Management of farm fish ponds.

Alabama Polytech. Inst. Agric. Exp. Sta., Bull, 254 : 1-30.

TOTH, S. J. AND D. N. RIEMER 1968. Precise chemical control of algae in

ponds. / . Amer. Water Works Assoc, 60 : 367-371.
TUCKER, C. S. AND C. E. BOYD 1978. Consequences of periodic applications
of copper sulfate and siraazine for phytoplankton control in catfish ponds.
Trans. Amer. Fish. Soc, 107 : 316-320.
 7
WATER ANALYSIS

The purpose of this chapter is to present methods of water

and mud analysis that are frequently employed in fish culture.
These include Secchi disc visibility. pH, total alkalinity, total
hardness, dissolved oxygen, carbon dioxide, chemical oxygen
demand, ammonia, nitrite, chlorophyll a, particulate organic
matter, hydrogen sulphide and lime requirement of muds. Also
included is a method for neutralizing value of limestone.
WATER SAMPLING
Water samples for dissolved oxygen or carbon dioxide analysis
must be collected so that they do not come in contact with air.

Fto. 7.1. Water sampler useful for collecting water from depths of
up to 2 m.
72
Van Dorn, Kemmerer or sewage samplers, which may be
purchased from scientific supply houses, are most commonly
employed for taking samples for dissolved gas analysis. Samples
for other variables may be dipped from the surface with open
containers. Samplers may be constructed from inexpensive
materials for securing samples from greater depths (Figs. 7.1
and 7.2). Of course, water collected with samplers of the types
shown in Figs. 7.1 and 7.2 would be contaminated with air and
unfit for dissolved oxygen or carbon dioxide analysis.

rubber stopper

weight

FIG. 7.2. A weighted bottle water sampler.

SECCHI DISC VISIBILITY

A Secchi disc is 20 cm in diameter, painted with black and

white quadrants and attached to a calibrated line (Fig. 7.3).
The disc is weighted on the underside with a lead plate so that it
will sink readily. Secchi discs may be purchased from scientific
supply houses or constructed from sheet metal, plexiglass or
masonite. A Jlat paint should be used to prevent glare. A
 73
suitable alternative to attaching the disc to a calibrated line is to
attach it from its centre to a vertical metre stick. Secchi disc
visibilities seldom exceed 100 cm in productive fish culture
systems, so measurements will seldom be limited because of the
length of the metre stick.

weight

groduoted rope

FIG. 7.3. A Secchi disc.

Secchi disc visibility is not a suitable estimate of plankton

unless plankton is the primary source of turbidity. An experi-
enced observer can readily distinguish between plankton
turbidity and other forms of turbidity. However, the novice
must remember that plankton blooms are not always green.
Plankton blooms may also impart yellow, red, brown or black
colouration to water. Usually plankton organisms are large
enough that their particulate nature is obvious if water and its
contents are viewed against a white background.
To obtain the secchi disc visibility, lower the disc into the
water until it just disappears and record the depth. Lower the
disc a little more and then raise it until it just reappears and
record the depth. In making these measurements, view the disc
from directly above. The average of the two depths is the Secchi
disc visibility. Conditions for taking Secchi disc measurements
should be standardised. A good practice is to make measure-
ments between mid morning and mid afternoon. The water should
be calm and the sun behind you.
74

TOTAL ALKALINITY

The amount of acid required to titrate the bases in water is a

measure of the alkalinity of water. A number of bases occur in
water, but total alkalinity results primarily from bicarbonate and
carbonate. For the determination of total alkalinity, a water
sample is titrated to pH 4.5, the methyl orange end point, with
standard acid. The amount of acid required for the titration is
equivalent to the alkalinity. Alkalinity is expressed as equivalent
calcium carbonate.

Reagents
Methyl orange indicator solution: Dissolve 0.05 g of methyl
orange in 100 ml of distilled water.
Standard sodium carbonate, 0-0200iv*; Dry a few grams of
NaaCOa (primary standard) at 140°C and cool in a desiccator.
Dissolve 1.0600 g of the Na2COs and dilute to 1,000 ml in COa-
free distilled water. Boil distilled water for 10 to 15 minutes to
expel COj and cool before using.
Standard sulphuric acid solution : Prepare a H a S0 4 solution
of approximately Q1N by diluting 2.8 ml of concentrated H a S0 4
to 1,000 ml with COa-free distilled water. Dilute 200 ml of
0.1N H a S0 4 to 1,003 ml with COa-free distilled water. This
solution is approximately 0.02N, but it must be carefully
standardised to determine its exact normality. To standardise,
pipette 10 ml of 0.0200N Na a C0 3 into a 250 ml beaker. Add
90 ml of COa-free distilled water and 4 to 8 drops of methyl
orange indicator solution. Titrate over a white surface to the
methyl orange end point with the sulphuric acid solution. At
end point, one drop of acid will change the colour of methyl
orange from yellow to faint orange. Calculate the normality of
the sulphuric acid with the following equation :
NV = N ' V
Where N = normality of sodium carbonate solution ;
V = volume in millilitres of sodium oarbonate solution ;
N' = normality of sulphuric acid solution ;
V = volume in millilitres of sulphuric acid solution.
 75

Procedure
Add 4 to 8 drops of methyl orange indicator solution to a
100 ml sample and titrate with standard sulphuric acid solution
until the colour of the solution changes from yellow to faint orange.
Measure the volume of sulphuric acid. Calculate total alkalinity
with the following equation :
Total alkalinity (mg/litre as CaC08) = (T) (NHS0»00°)
where T = volume in millilitres of sulphuric acid ;
N = normality of sulphuric acid ;
S = volume in millilitres of sample.

TOTAL HARDNBSS

The concentration of calcium plus magnesium expressed as

equivalent calcium carbonate is the total alkalinity. Calcium
and magnesium ions are titrated with the complexing agent
ethylenediamine tetracetic acid disodium salt (EDTA) to form
the stable complexes CaEDTA and MgEDTA. The end point of
the titration is signalled with an indicator called eriochrome
black-T.

Reagent
Buffer solution : Dissolve 67*5 g of NH4C1 in |570 ml of
concentrated NH4OH. Dilute to 1,000 ml in a volumetric flask
with distilled water.
Eriochrome black-T indicator: Dissolve 4-5 g of hydroxyla-
mine hydrochloride and 0-50 g of eriochrome black-T in 100 ml
of 70 % ethanol. Prepare fresh every 2 to 3 months.
Standard calcium solution, 0-010 M: Transfer 1000 g of
anhydrous CaC03 to a 1,000 ml beaker. Add 1: 1 HCl slowly
to dissolve the CaC08 and dilute to about 200 ml with distilled
water. Boil for 5 to 10 minutes to expel carbon dioxide, cool
and adjust to pH 7, as determined with a pH meter, with 3 N
NH4OH. Transfer to a 1,000 ml volumetric flask and dilute to
volume with distilled water.
76
Standard EDTA solution : Dissolve 4*00 g EDTA disodium
salt and 100 mg of MgCl, - 6HaO in distilled water and dilute to
1,000 ml. The solution must be standardised against the
standard calcium solution. Pipette 10 ml of the standard
calcium solution into a 250 ml beaker and add 90 ml of distilled
water. Titrate the calcium solution with EDTA solution
according to the procedure given below. Compute the molarity
of the EDTA solution with the equation : NV » N'V.

Procedure
Measure a 100 ml water sample into a 250 ml Erlenmeyer
flask. Add 2-0 ml of the buffer solution and mix. Add 8 drops
of eriochrome black-T indicator and titrate with the EDTA
solution. At the end point, the solution will change from wine-
red to pure blue. Calculate the total hardness with the equation :
Total hardness (mg/litre as CaCOa) = ( T ) ( M ) 0°°» 10°)
where T = volume in millilitres of EDTA solution ;
M = molarity of EDTA solution ;
S = volume in millilitres of sample.

Comment
For samples high in hardness, e.g., sea water, dilute a 1-0 to
10-0 ml water sample to 100 ml with distilled water. Use the
actual volume of sample in the calculation.

HYDROGBN ION (pH)

Various types of indicator papers and solutions have been

used to measure pH. However, the only reliable technique is the
electrometric pH meter. The manufacturer's instructions should
be consulted for use of a pH meter. Before making pH measure-
ments, carefully calibrate the meter with a pH 7 buffer.
However, this procedure does not verify that the meter will read
other pH values correctly. A second buffer, pH 5 if samples are
expected to be acidic or pH 9 if samples are expected to be basic,
should be used to determine if the pH meter will read a second
pH correctly after it has been calibrated at pH 7.
 ft
DlSSOLVBD OXYGBN

In the basic Winkler procedure, a sample of water is treated

with manganous sulphate, potassium iodide and sodium
hydroxide. Under highly alkaline conditions, the manganous ion
is oxidised by molecular oxygen to manganous dioxide, a brown
precipitate.
Mn8+ + 20H" + l/20a - • MnOg + H 8 0
Notice that only one half of the oxygen in manganous dioxide
came from molecular oxygen. The formation of a white
precipitate will occur even in the absence of oxygen since the
manganous ion will form manganous hydroxide, a white
precipitate.
Mn*+ + 20H~ -*» Mn (OH),
Next, sulphuric acid is added to the sample to dissolve the
precipitate and produce acid conditions for the oxidation of
iodide to iodine by manganous dioxide according to the following
reaction:
MnO, + 21" + 4H+ -+ Mn*+ + I, + 2H,0
The quantity of I, released is proportional to the amount of Oa
originally present. One half of a molecule of O, resulted in the
release of one molecule of iodine (I2). The amount of I, is
estimated by titration with standard sodium thiosulphate. A
starch indicator is used to determine the end point of the
titration. As long as iodine is present, the solutionis blue.
When all the iodine has been titrated the solution becomes
colourless.
I8 + Starch - It + 2NajS803 . 5HjO -*
(Blue)
Na,S406 + 2NaI + 10H.0 + Stareh
(colourless)
The amount of iodine is used to calculate the original DO
concentration.
Reagents
Manganous sulphate solution: Dissolve 364 g of MnS04HtO
in distilled water, filter and dilute to lf000 ml in a volumetric
flask.
n
Alkali-iodine-azide solution : Dissolve 500 g of NaOH and
150 g of KI in distilled water and dilute to 1,000 ml in a
volumetric flask. Dissolve 10 g of NaN8 in 40 ml of distilled
water and add to the NaOH-KI solution.
Sodium thiosulphate solution : Dissolve 6-3 g of Na2S2Os-5H20
in freshly boiled and cooled distilled water and dilute to 1,000 ml
in a volumetric flask. There is no need to weigh the Na a S 2 0 3 .
5H 2 0 with more precision since the resulting solution must be
standardised. Add 5 drops of chloroform as a preservative.
This reagent must be restandardised every few days and stored in
the dark.
Concentrated sulphuric acid: Analytical reagent grade.
Sulphuric acid solution, 10 percent: Add 5 ml of concen-
trated H 2 S0 4 to 45 ml of distilled water.
Potassium dichromate solution, 00250 N : Dry 2 or 3 g of
K2Cr207 at 105°C and cool in a desiccator. Dissolve 0-6129 g of
K 2 Cr 2 0 7 and dilute in a volumetric flask to 500 ml with freshly
boiled and cooled distilled water.
Starch indicator: Add 2 g of soluble starch to 100 ml of
distilled water in a 250 ml beaker. Heat while stirring until
transparent and add 0-5 ml of formalin as a preservative.

Procedure
Standardisation of sodium thiosulphate solution: Dissolve
2 g of KI in a H2SOB solution. Use volumetric pipette to measure
10 ml of 0-025 N K a Cr a 0 7 into the flask and place the flask in
the dark for 5 minutes. Dilute to 250 or 300 ml with distilled
water. Titrate with sodium thiosulphate solution until a pale
straw colour is reached. Add 8 drops of starch indicator and
titrate until the blue colour of the starch suddenly disappears.
Record the volume of sodium thiosulphate used. Calculate the
normality with the equation: NV = N ' V .
Preparation of sample for analysis : If a bottle train type
sampler is used, the BOD bottle must be removed and stoppered
carefully to prevent the introduction of air bubbles. If a Van
 rt
Dorn or Kemmerer bottle is used, introduce the water through a
tube which discharges at the bottom of the BOD bottle, allow the
BOD bottle to overflow for two or three exchanges of water and
stopper it carefully to prevent air bubbles. Samples must be
analysed for DO without delay.
Fixation of dissolved oxygen and liberation of iodine: To
a sample in a 300 ml BOD bottle, add 2 ml of manganous
sulphate solution and 2 ml of alkali-iodine-azide solution. These
reagents may bs added with a measuring pipette. Introduce the
reagents below the surface of the sample and stopper with care to
prevent air bubbles. Mix the solution in the bottle by rapidly
inverting it twenty times and then let the sample stand until a
precipitate settles to the bottom half of the bottle. Mix again by
inverting the bottle several times and let the precipitate settle.
Add 2 ml of concentrated HaS04 with a measuring pipette,
stopper the bottle and invert several times to dissolve the
precipitate.
Titration of iodine: To compensate for overflow of the
sample during the addition of the reagents, a 101 ml sample is
taken for titration. Calibrate a 100 ml graduated cylinder
by measuring 101 ml into it and placing a suitable graduation
at the appropriate position above the manufacturer's 100 ml
graduation. Shake the BOD bottle and then measure 100 ml
into a 250 ml beaker. Titrate with standard sodium thiosulphate
solution to a pale straw colour. Add 8 drops of starch indicator
solution and titrate until the blue colour disappears. Record
the volume of sodium thiosulphate used. Use the following
equation to calculate the DO concentrations :
Dissolved oxygen (rag/litre) . <T) (N) (8,000)
where T = volume in millilitre of sodium thiosulphate;
N = normality of sodium thiosulphate;
S = volume in millilitres of sample.

Comments
Consult a more detailed water analysis manual if there is any
question about interference. A sample can be preserved for up
80
to 8 hours by adding 0-7 ml of concentrated H 2 S0 4 and 1 ml of
2% NaN 3 and storing the sample in a sealed BOD bottle at 20°C
or less. In completing the procedure, add 3*0 ml of alkali-
iodide-azide solution rather than the usual 2-0 ml. Dissolved
oxygen may also be fixed at the sampling site by the addition of
manganous sulphate and alkali-iodide-azide solution and the
sample carried to the laboratory within 2 or 3 hours for prompt
completion of the DO determination.
Standard phenylarsine oxide (PAO) may bs purchased (Hach
Chemical Company, Loveland, Colorado) and used instead of
sodium thiosulphate in the titration of DO. PAO does not have
to be continually restandardised because it is stable for several
months.

CHLOROPHYLL a

The phytoplankton in pond water is concentrated by filtration

through a membrane filter. The pigments contained in the
phytoplankton are extracted in acetone and the concentration of
chlorophyll a determined spectrophotometrically. A close
relationship usually exists between the concentration of
chlorophyll a in water and the total abundanoe of phytoplankton,

Special apparatus
The following special items are needed : Millipore filters
(Type HA, 47 mm, 0-45 micron pores), millipore filtration
apparatus, small electric drill, tissue grinder with 10 ml
chamber and Teflon pestle (A. H. Thomas Co., Philadelphia,
PA., Cat. No. 3431-E15), centrifuge and spectrophotometer.

Reagents
Acetone 90% : Add 50 ml of distilled water to 450 ml of
reagent grade acetone.
Magnesium carbonate suspension 1 % : Place 10 g of powdered
MgC0 3 in a 100 ml volumetric flask and dilute to volume with
distilled water. Only a small amount of the MgCO, will dissolve.
 fill
Procedure
Place a millipore filter on the filter holder and attach the
funnel. Shake the MgCO„ suspension and pipette 1.0 ml over the
filter. Apply vaccuum to remove the liquid from the filter.
Transfer 50 or 100 ml of the well mixed sample to the funnel.
After the sample hasfilteredthrough, remove the Millipore filter
and trim away the edges which are not coated with residue.
Crumple the filter and place it in the tissue grinder. Add
2 ml of 90 % acetone and grind for 1 minute, then add 8.00 ml
of 90% acetone and grind for 30 seconds. Transfer the contents
of the tissue grinder tube to a 50 ml Erlenmeyer flask, stopper
and refrigerate in the dark for 1 hour. Pour the acetone extract
into a 15 ml test tube and centrifuge at 2,000 to 3,000 revolutions
per minute (rpm) for 10 minutes. Decant the acetone extract
, into a cuvette and centrifuge at low speed (300 to 500 rpm) for
five minutes. Measure the absorbance of the acetone extract at
665 nm and again at 750 nm with a spectrophotometer set at 00
absorbance with 90% acetone. Calculate the chlorophyll a
concentration from the equation :
Chlorophyll a in F g/litre= 11-9 (Am - AT50) -£ x i s p
where A685 = the absorbance at 665 nm;
A7B0 = the absorbance at 750 nm;
V = the acetone extract in millilitres
L = the length of light path in the speotrophotometre
in centimetres;
S = the volume in millilitres of sample filtered.

PARTICULATE ORGANIC MATTBR

Glassfibrefiltersare available which will retain most of the

particles in water which are greater than 1 micron in size. When
pond water is passed through such a filter, essentially all of the
living plankton and much of the particulate matter is retained.
The filter and residue are then dried and weighed. Next, the
filter and residue are ignited, cooled and weighted again. The
weight loss on ignition is taken as the particulate organic matter
content of the sample.
6
82

Special apparatus
The following special items are required : glass fibre filtration
apparatus, 47 mm Gelman Type A-E glass fibre filters or
equivalent and a muffle furnace.

Procedure
Prepare glass fibre filters by soaking them in distilled water
for 24 hours, drying them and igniting them in a muffle furnace
for 20 minutes at 550°C. Place a glass fibre filter on the filter
holder, attach the funnel and transfer a well-mixed and accurately
measured sample of water to the funnel with a graduated
cylinder. The sample volume must be large enough so that the
residue retained on the filter will be heavy enough to weigh. A
volume of 250 ml is usually adequate for pond waters. Apply
vacuum and after the sample has drained through the filter,
wash the filter and residue with 20 ml of distilled water. Dry
the filter and reside at 103°C, cool in a desicoator and weigh.
Ignite the tarred filter and residue at 550°C in a muffle furnace
for 20 minutes, cool in a desiccator and weigh again. The
weight loss represents particulate organic matter. Use the
equation below to calculate the concentration of particulate
organic matter.

Particulate organic matter (mg/litre) «a (B - A) —^—

where B = the weight of the filter and residue in milligrams
before ignition;
A =» the weight of the filter and residue in milligrams
after ignition.
S = volume of sample in millilitres.

CARBON DIOXIDB

Water which has a pH above 8-3 does not contain appreciable

carbon dioxide. Therefore, the amount of base required to raise
the pH of a water sample to the phenolphthalein end point is
approximately equivalent to the carbon dioxide content of the
sample.
 **

fceagehtS

Phenolphthalein indicator solution: Dissolve 0*5 g of

phenolphthalein in 50 ml of 95% ethyl alcohol and add 50 mg of
COj free water. Add 004554 N sodium oarbonate dropwise
until a faint pink colour appears.

Standard sodium corbonate 0-054 N: Dry a few grams of

Na a CO s and dilute to 1,000 ml with CO a free distilled water.
This solution must be prepared fresh daily.

Procedure

To minimise exposure to air, siphon a portion of the sample

into a 100 ml graduated cylinder through a flexible tube which
discharges at the bottom of the cylinder. Let 50 to 75 ml of
water overflow from the cylinder and remove tube. Remove
excess sample from cylinder with a pipette. Add 4 drops of
phenolphthalein indicator solution. If the sample turns pink,
CO, is absent. If sample remains colourless, titrate with Nat
COa while stirring gently with a stirring rod. A faint pink
colour that remains for 30 seconds marks the end point.
Calculate COa as follows t
Carbon dioxide (mg/litre) - ( T ) (N) (22,000)
where T = volume in millilitres of sodium carbonate j
N = normality of sodium carbonate;
S = volume in millilitres of samples.

CHBMICAL OXYGBN DBMAND

In this modification of the COD procedure, no heat other

than that produced by the dilution of concentrated sulfuric acid is
applied to the sample. In acid solution, potassium dichromate
oxidizes organic matter to carbon dioxide and water. The
amount of potassium dichromate consumed is equivalent to the
quantity of readily oxidizable organic matter in the sample. r '
84

Reagents
Potassium dichromate solution l.OOOJV: Dry primary standard
grade K,Cr t 0 7 at 103°C for 2 hours and cool in a desiccator.
Dissolve 49.036 g of K,Cr s O, in distilled water and dilute to
1,000 ml.
Potassium dichromate solution 0-0250JV": Dilute 25.00 ml of
1,000 N K8CrjG7 and 100 mg of sulfamic acid to 1,000 ml with
distilled water.
Ferrous ammonium sulphate solution : Dissolve 9.8 g of Fe
(NH4) (S04) • 6H,0 in distilled water and add 20 ml of concen-
trated H a S0 4 . Cool, dilute to 1,000 ml and store in the dark.
Standardise daily as follows. Dilute 10 ml of 0.0250 N
K,Cr,0 7 to about 45 ml with distilled water in an Erlenmeyer
flask and add 30 ml of concentrated sulphuric acid. When cool,
add 2 or 3 drops of ferroin indicator and titrate with ferrous
ammonium sulphate as described in the procedure below.
Calculate normality as : NV = N ' V .
Ferroin indicator: Dissolve 1.8877 gof 1, 10-phenathroline
raonohydrate and 0.7 g of Fe S04-7HaO in 100 ml distilled
water.
Other reagents: Reagent grade silver sulphate, mercuric
sulphate and concentrated sulphuric acid.

Procedure
Clean glassware with H g S0 4 -Na a Cr a 0 7 cleaning solution and
rinse thoroughly in distilled water. Pipette a 20 ml sample
and 10 ml of the standard dichromate solution (0O25N) into
a 125 ml Erlenmeyer flask. A reagent blank is prepared from
distilled water and treated exactly as the samples. Add 30 ml
concentrated sulphuric acid, 0 4 g Ag S0 4 crystals and enough
HgS0 4 crystals to maintain an HgS0 4 : chloride ratio of 10.
Swirl. Cover flasks with clean cover glasses and let stand for
30 minutes. Dilute with 75 ml of distilled water. Add 2 or 3
drops of ferroin indicator and titrate with standard ferrous
ammonium sulphate (0.O25N). Initially, samples will vary in
 if
colour (yellowish orange to blue-green), but just before the end
point all samples will turn blue-green. At the end point, a*
single drop of titrant causes the colour to change from blue-
green to red-brown. If samples are high in organic matter, use
more concentrated solutions of potassium dichromate and
ferrous ammonium sulphate. Calculate the COD with the
following equation:
COD in mg/litre - 0 > - - A ) ( N ) (8.000),

where B =» millilitres of ferrous ammonium sulphate (PAS) used

in the titration of the reagent blank;
A a millilitres of FAS used in the titration of the sample;
N =. normality of the FAS;
S - volume in millilitres of sample.

NlTRITB

The colorimetric methods generally used for nitrite employ

diazotising reagents. Nitrite reacts with these reagents in acidic
solution to form diazonium salts. The diazonium salts are
coupled with amino or hydroxy] groups of aromatic compounds to
form coloured azo compounds. The method given here employs
sulphanilamide as the diazotising reagent and N-(l-naphthyl)-
ethylenediamine as the coupling reagent. The azo compound is
bright pink and a concentration of Q.01 mg/litre NO„-N produces
a disceraable colour.

Instrument
A spectrophometer capable of operating at 543 nm is required.

Reagents
Diazotising reagent: Add 5 g of sulphanilamide and SO m]
of concentrated hydrochloric acid to 300 ml of distilled water in
a 500 ml volumetric flask. Stir to dissolve and dilute to volume.
Coupling reagent: Dissolve 500 mg of N- (1-naphthyl)-
ethylenediamine dihydrochloride in 500 ml of distilled water.
86
Store in a dark bottle out of the light. This reagent gradually
becomes dark brown and must be prepared fresh every 2 to 4
weeks.

Standard nitrite-nitrogen solution 1-00 mg/litre: Dissolve

0.4925 g of NaNO, in 1,000 ml of distilled water. This solution
contains 100 mg/litre of NO,-N. Pipette 10 ml of the 100 mg/
litre NO a -N solution into a 1,000 ml volumetric flask and dilute
to volume with distilled water to give a 1.00 mg/litre NO,-N
solution. These solutions deteriorate rapidly.

Procedure

Development and measurement of pink colour : Filter the

water sample through Whatman No. 42 or equivalent, filter
paper. Measure 50 ml of water into a 100 ml beaker. Add
1.0 ml of diazotising reagent, stir and allow 2 to 4 minutes (no
longer) for reaction. Add 1.0 ml of coupling reagent and stir.
Let the solution stand for 10 minutes to form the azo compound,
transfer to a 1 cm cuvette and measure the pink colour spectro-
photometrically at 543 nm. Use a reagent blank to set the
spectrophotometer at 0-0 absorbance (100% transmittance).
Obtain the concentration of nitrogen from a calibration graph.

Calibration graph: Use the standard NO a -N soutlion

(1.00 mg/litre) to prepare a series of NOa-N concentrations. The
concentrations listed below are suitable for use with a spectro-
photometer with a 1 cm light path length.
Nitrite-Nitrogen Millilitres of 1.00 mg/litre nitrite-
(mg/litre) nitrogen standard diluted to 100 ml
0.00 0.00
0.02 2.00
0.04 4.00
0.06 6.00
0.08 8.00
0.10 10.00
0.15 15.00
0-20 20.00
 17
The aliquots of standard nitrite solution must be transferred
to volumetric flasks with volumetric pipettes. Add reagents to
develop the colour. Use the 0-0 mg/litre solution to set the
spectrophotometer to 0 0 absorbance or 100 per cent transmittance
and evaluate the pink colour of the other solutions at 543 nm.
Plot the data to obtain a calibration graph.

DETERMINATION OF AMMONIA

Reagents

All the reagents are prepared using ammonia free distilled

water.
Phenol-alcohol solution : Dissolve 10 g of reagent grade
phenol in 100 ml of 95% v/v ethyl alcohol U.S.P.
Sodium nitroprusside 0.5%: Dissolve 1 g of sodium nitro-
prusside in 200 ml of water.
Alkaline solution : Dissolve 100 g of trisodium citrate and
5 g of sodium hydroxide in 500 ml of water.
Sodium hypochlorite solution : Use a solution of commercial
hypochlorite which should be atleast 1.5 N.
Oxidizing solution : Mix 100 ml of sodium citrate solution
and 25 ml of hypochlorite solution'and use the same day.'
Stock standard solution : 0100 g Ammonium sulphate
(A.R. grade) in 1000 ml distilled water. 1 ml = 1-5 pg at N.

Method
For natural waters, the procedure consists of the successive
addition of 2 ml of phenol solution, 2 ml of sodium nitroprusside
solution and 5 ml of oxidising reagent to 50 ml of sample,
mixing thoroughly after each addition. The color is allowed to
develop at room temperature (22-27°C) for 1 hr and the
absorbance recorded at 6400 A in a spectrophotometer with a
10 cm length cuvette. All the glassware used must be cleaned by
washing initially with warm dilute hydrochloric acid and rinsing
thoroughly with distilled water.
88

Procedure and calculation

Dilute the standard stock solution to get working standards
of 1-5 ; 3-0 ; 4-5 ; 6-0 and 9-0 fig at N concentrations. Measure
the absorbance at a wavelength at 6400 A in a spectrophotometer
(10 cm cell) and draw a calibration graph. Compare the
absorbance of the given sample and calculate the ammonia
concentration from the calibration graph.

HYDROGEN SULPHIDB*

Sulphide is found in anoxic waters, where it is formed by

microbiological reduct ion of sulphate ions. The method described
here, depending on the formation of methylene blue from
dimethyl p-phenylene diamine, is a simple application of a well
established colorimetric method for sulphide.
The method is based on the following principle. The acidified
sample is allowed to react with dimethyl p-phenylene diamine,
with ferric ions as catalyst. A complex oxidation and substitution
takes place, resulting in the quantitative incorporation of any
sulphide-sulphur present into a heterocyclic dye called methylene
blue. The absorption of light by the sample is measured before
or after dilution in 1, 5 or 10-cm cells. The method is as nearly
sensitive as is theoretically possible and is for direct determi-
nation applicable to concentrations upto about 100//*gat/l or
about 3-2 mg/1. At higher concentrations of hydrogen sulphide
the sample has to be diluted with oxygen-free distilled water
prior to the analysis.

Reagents
N, N-dimethyl p-phenylene diamine dihydrochloride, p.a.
(CH,),NC6H4 • NH 8 2HC1 (1, 4 ) : 1 g is dissolved in about 6 N
hydrochloric acid to 500 ml. This acid may be prepared by
diluting concentrated HC1 (37 percent, sp.gr. 119) with an
equal amount of distilled water.
* Reproduced with the kind permission of FAO, Rome from FAO Fisheries
Technical Paper No. 137 ; 132-135.
 89
Ferric chloride, p.a., FeCls : 8 g is dissolved in about 6 N
hydrochloric acid (prepared as above) to 500 ml.

Oxygen-free distilled water : A suitable volume of distilled

water is boiled for 30 to 60 minutes. Nitrogen gas may be bubbled
through the water during the boiling. As the water cools down
to room temperature, nitrogen gas must be bubbled through.
This water is difficult to store, so it should be prepared just
before use.

Sulphide stock solution : A solution of sodium sulphide is

prepared with oxygen-free distilled water. Crystals of
Na2S • 9H,0 p.a. are quickly washed with distilled water by
squirting from a washing bottle. The crystals are dried with
filter paper and placed in a pre-weighed, glass stoppered,
weighing glass. 0-750 g is weighed on an analytical balance and
dissolved in oxygen-free distilled water (added with aid of a
siphon) to 1000 ml in a volumetric flask.

Sulphide working solution : Siphon a suitable volume of

oxygen-free distilled water into a 500 ml volumetric flask. Into
this is pipetted 25 ml of the sulphide stock solution and oxygen-
free distilled water is added to the mark. This solution contains
approximately 5 /ttg/ml as sulphide, S* - (about 0-156 /*g at/ml).
For the greatest accuracy, the concentration is detefmined by
titration. The solution is stable for only 15 - 30 minutes.

Sodium thiosulphate solution, 0.02N .- 5g NaaSa08.5HaO p.a.

is dissolved in 1000 ml distilled water. Add 5 ml isobutyl
alcohol before diluting to the full volume of the volumetric
flask.

Potassium hydrogen todate solution: Weigh out accurately

1.2998 g KH(IO„)a p.a. dried at 105°C for one hour. Dissolve
in distilled water and dilute to 1000 ml. This solution is 0.04 N
and very stable.

Sulphuric acid (1 + 1) solution : One volume of acid is care-

fully mixed with one volume of distilled water under constant
cooling and mixing.
90
Starch solution: 1% or Thyodene indicator as for the
dissolved oxygen determination.
Potassium iodide, KI, p.a. crystals.

Equipment
Winkler bottles, Spectrophotometer or filterphotometer with
filter at or close to 670 nm. 1 cm and S cm or 10 cm cells and
automatic syringe pipettes.

Calibration
Standardisation of the thiosulphate, determination of the
factor ( / ) : Add 1 g pottassium iodide and 2 ml sulphuric acid
(1 + 1) to an Brlenmeyer flask with a ground stopper containing
approximately ISO ml distilled water. Mix well and add 10 ml
of the pottassium hydrogen iodate solution by means of a
pipette and stopper the flask. Swirl gently and allow the iodine
liberation to proceed (away from direct sunlight)for 2-4 minutes.
Titrate the iodine with the thiosulphate solution to a pale straw
colour. Add two drops of the starch solution as indicator and
complete the titration until ths disappearance of the blue colour.
A blank is run to check the reagents. This is done by repeating
the process, but omitting the hydrogen iodate. If v is the milli-
litres of thiosulphate consumed, then

10 - 0.04 = f. v . 0.02 and f = 2?

v
The mean value of (f) should be calculated after at least
three runs, differing not more than 0.05 ml.

Standardisation of the sulphide working solution: This is

carried out within minutes after preparation of the working
solution and at the same time as the preparation of the photo-
metric standard samples.

Take six Brlenmeyer flaskes with ground stoppers and add to

each about 10 ml distilled water and 1-2 g potassium iodide.
Pipette into each flask 10 ml hydrogen iodate solution. Add
 91
1.0 ml sulphuric acid (1 + 1) into each flask. Into three of the
flasks pipette 50 ml of, the sulphide working solution and to the
other three add about 50 ml distilled water. Set all flasks aside
in a cool place whilst the colorimetric standardisation is com-
menced and then titrate the contents of the flasks with thio-
sulphate using starch indicator.
A = mean of the titrations of three solutions with no added
sulphide, in ml.
B = mean of the titrations of the three solutions containing
sulphide, in ml.
f = factor of the thiosulphate.
(the individual titrations constituting a triplicate should agree to
within 0.05 ml).
H . S m l / l ^ 2 ^ 1 ^ ^ )

To convert this to /xgat/ml H,S (or rather p moles, if the

result is reported as hydrogen sulphide instead of sulphide, Sa~),
the number is divided by 22.140:

Photometric standard samples: From the working solution

the following standard series is prepared.
To 100 ml measuring flasks or volume calibrated Winkler
bottles the following volumes of working solution are added by
means of a pipette or burette (Table 7.1):

TABLE 7.1

20 ml = 100 /xg Sa~ = 31.2 ng at/1 S*'

16 ml = 80 „ „ = 25.0 „
12 ml 60 „ „ - 18.7
8 ml = 40 „ „ => 12.5
4 ml = 20 „ ,, = 6.3 ,, „
0 ml = 0 „ „ = 0.0 „
92
These concentrations correspond to the solution with 5 jig/ml.
Thus they have to be corrected according to the value found by
titration.
If Winkler bottles are used the concentration values given
above have to be recalculated accordingly for the calibration
graph described below.
With the aid of a siphon the bottles are filled up with oxygen-
free distilled water to 100 ml mark or in case of calibrated
Winkler bottles, to the neck. In the latter case a correction for
the volume has to be made for each bottle.
As soon as a bottle is filled up, 1 ml of each reagent is added
with an automatic syringe pipette and the contents of the bottle
are mixed well. After 60 minutes, the samples are measured
against the blank, at 670 nm in 1 cm and/or S cm cells, as suitable.
From the results a calibration graph for each of the cell lengths
is prepared on millimeter paper. The graph should be a straight
line and go through origo.
If higher concentrations are analysed, the graph will start
deviating from the straight line at 40 - 50 fj,g at/1 (about 1.3 -
1.6 mg/1). The exact point for the start of the deviation
depends on the quality of the amine solution. However, if this
is taken into account when calculating the analytical results, the
graph — and thus the working range of the method — can be
extended upto about 100 /*gat/l (about 3" 2 mg/1).

Procedure
The samples must be taken with plastic water samplers. If
these are not available, metallic ones may be used provided that
the inner surfaces are plastic-coated.
An ordinary Winkler bottle is filled with the sample in
exactly the same manner as described for the dissolved oxygen
determination. The reagents diamine and ferric chloride are
immediately added with pipettes (preferably automatic syringe
pipettes) to the sample. Let the pipette tips extend deep into the
sample bottle. The stopper is now inserted avoiding air bubbles.
 93
The blue colour starts developing in a few minutes and the
sample is ready for measurement in a photometer after 30
minutes. However, if the sample contains high concentrations
of hydrogen sulphide, one hour must be allowed for full colour
development. The colour intensity may be regarded as constant
for at least 24 hours.
The colour intensity of the sample is measured against distilled
water (or compensated for by reagent blanks) at 670 nm using 1
or 5 cm cells as required.
If the sample contains higher concentrations of hydrogen
sulphide than 100 ^gat/1 (3.2 mg/1) it has to be diluted prior to
the analysis. This is done by pipetting a suitable volume of
the sample into a measuring flask or Winkler bottle containing
some oxygen-free distilled water. The pipette tip should extend
below the surface of the water. Then more of the siluting water
is added by means of a siphon upto the calibrated volume of the
flask or bottle. The reagents are added and the sample is then
thoroughly mixed. Account has to be taken of the silution
factor when calculating the result of the analysis.
Note: It is difficult to prepare a standard solution of
sulphide with a high degree of accuracy. Therefore a systematic
error may be included in the analysis. Using the method
described, this error will be below 2 %.
It seems impossible to obtain a diamine that is not more or
less discoloured. However, this does not seem to affect the
results. Analysis done at the Fishery Board of Sweden with
a one year old brownish diamine solution produced results
which differed only slightly from the results obtained with a
freshly prepared solution.
According to Strickland and Parsons (1968) it may be
necessary to compensate the absorption value of each sample
for the absorption value of a reagent blank. This latter value
is obtained by adding reagents to filtered surface water and
measuring this against the same filtered water containing no
reagents. The absorption value should not exceed 05 in a
10 cm cell and should preferably be less than 0-2S.
<M
LIME REQUIREMENT

Boyd (2, 3) altered a lime requirement procedure for agricul-

tural soils (1) so that it could be used to estimate liming rates
for fish ponds. Boyd (2) found that total hardness and total
alkalinity of pond waters exceeded 20 mg/ litre when the base
unsaturation of muds was 0.2 or less. Furthermore, there was
a strong correlation between the base unsaturation of pond
muds and their pH values and a mud pH of 6.0 corresponded
to a base unsaturation of 0.2. Therefore, if the pH of
a mud was known, the base unsaturation could be computed
from the regression equation relating pH and base unsaturation
of muds. The reduction of pH in a buffer solution caused by a
known weight of dry mud provided an estimate of total exchange
acidity. Neutralization of the total exchange acidity would give
a base unsaturation of 0.0. Therefore, the amount of exchange
acidity (in milliequivalents) which must be neutralized to lower
the base unsaturation to 0.2 — this corresponds to 20 mg/1 total
hardness and alkalinity in pond water—was calculated as
follows :
Acidity to be = exchange acidity desired change in
neutralized initial base unsaturation base unsaturation
Boyd and Cuenco (4) determined that agricultural limestone
reacted to a depth of approximately 15 cm in pond muds over a
2 year period and that air-dry weight of the upper 15 cm layer
of mud in ponds averaged 1,400,00 kg/ha. The amount of
acidity to be neutralised as estimated for the sample may be
expanded to give the amount of acidity to be neutralised in the
pond mud. The liming rate in kilograms per hectare of CaCOa
is calculated from the acidity. A liming factor of 1.5 is
multiplied by the liming rate, because agricultural limestone is
not 100 % effective in neutralising soil or mud acidity.
The method developed by Boyd (2) has proven effective and
it has been widely used. However, the procedure was developed
for ponds in Alabama and relationships between pH and base
unsaturation of muds differ geographically. For most accurate
results, the relationship between pH and base unsaturation of
muds should be determined for a region before the procedure is
 9$
Used. This is a difficult task that is often impractical. A simple
method developed by Pillai and Boyd (5) for determining the
lime requirement of pond muds that does not require data on
the relationship between pH and base unsaturation is presented.
This method involves measuring the total exchange acidity with
a buffer and calculating the liming rate necessary to provide a
base unsaturation of 0.0. Because this technique will always
provide a higher liming rate than necessary, the liming factor of
1.5 is omitted.

Reagents
Buffer solution : Dissolve 20 g p-nitrophenol, 15 g boric acid,
74 g potassium chloride and 10.5 g potassium hydroxide in
distilled water and dilute to 2,000 ml with distilled water.

Procedure
Weigh 20 g of air-dry mud that passess a G.85 mm screen into
a 100 ml beaker and add 40 ml of buffer. Stir intermittently for
1 hrand measure the pH to the nearest 0.01 pH unit with a
glass electrode. Each 0.1 unit change in pH represents 0.16
meq of acidity. If the equilibrium pH is less than 6.8, the
procedure must be repeated using half as much dry mud.
Calculate the lime requirement as follows :
Liming rate (kg CaCOs/ha) = pH change x 5,600

NEUTRALISING VALUB OF LIMING MATERIALS

The methods provides an estimate of the strength of a liming

material in neutralising acidity. Results are in terms of per-
centage of pure calcium carbonate.

Reagents
Standard hydrochloria acid, 1.000 N.
Standard sodium hydroxide, 1.000 N.
Phenolphtbalein indicator solution.
96

Procedure
Finely pulverize the liming material., Weigh 500 mg into a
500 ml Erlenmeyer flask. Add 25 ml of standard hydrochloric
acid —add slowly to prevent splattering. Boil briskly for 5
minutes. Heat on water bath for 3Qptninutes. Dilute to 100 ml
with distilled water and allow to cool. Add .a few drops of
phenolphthalein indicator and titrate with standard sodium
hydroxide to a faint pink colour. Calculate the neutralising
value as follows :
Neutralising value (*) = ^ z D J ^ m

where T = volume in millilitres of standard NaOH ;

N =3 normality of standard HCI;
S = sample weighjjn milligrams.

BIBLIOGRAPHY

ADAMS, F. AND C. E. EVANS 1962. A rapid method for measuring lime require-
ment of red-yellovrpodzolic soils. Soil $ci. Soc. Am. Proc., 26 ; 355-357.

BOYD, C. E. 1974. Linje requirements of Alabama fish ponds. AL Agr.

Exp. Sta., Auburn Univ.AL, Bull., 459 : 1-20.

1976. Lime requirement and application in fish ponds. In : T.V.R.

Pillay and W. A. Dill (^A^ Advances in Aquaculture. Fishing News (Books)
Ltd., Farham, England, pp. 120-122.

AND M. L. CUENCO 1980. Refinements of the lime requirement

procedure for fish ponds. Aquaculture, 2 1 : 293-299.

F.A.O. 1975. Manual of methods in Aquatic Enviroriiftjiit Research.

Part I. Methods for detection, measurement and monitoring of water
pollution. FAO Fisheries Tectmioal Paper No. 137 : 132-135.

PILLAI, V. K. AND C. E. BOYD 1984. A simple method for calculating liming

rates for fish ponds. Manuscript submitted to Aquaculture.
ZOLARZANO, L. 1969. Determination of Ammonia fn natural waters by the
phenol hypochlorite method. Limnol. Ocemogr., 14 : 799-801.
 Manuals of Research Methods issued under the Centre
of Advanced Studies in Mariculture, Central Marine Fisheries
Research Institute, Cochin.
1. Manual of Research Methods for Crustacean Biochemistry
and Physiology. CMFRI special Publication No. 7, 1981,
172 pp.
*2. Manual of Research Methods for Fish and Shellfish
Nutrition. CMFRI Special Publication No. 8, 1981, 125 pp.
3. Manual of Research Methods for Marine Invertebrate
Reproduction. CMFRI Special Publication No. 9, 1982, I
214 pp. I
*4. Approaches to Finfish and Shellfish Pathology Investi-
gations. CMFRI Sp.cial Publication No. 11, 1983, 43 pp.
5. Application of Genetics in Aquaculture. CMFRI Special
Publication No. 13, 1983, 90 pp.
6. Manual of Research Methods for Invertebrate Endocrino-
logy. CMFRI Special Publication No. 14, 1983, 114 pp.
7. Production and Use of Artemia in Aquaculture. CMFRI
Special Publication No. 15, 1984, 74 pp.
8. Manual on Marine Toxins in Bivalve Molluscs and General
consideration of Shellfish Sanitation. CMFRI Special
Publication No. 16, 1984, 100 pp.
9. Handbook on Diagnosis and Control of Bacterial Diseases in
Finfish and Shellfish Culture. CMFRI Special Publication I
No. 17, 1984, 50 pp.
10. Mariculture Research under the Centre of Advanced Studies
in Mariculture. CMF^I Special Publication No. 19, 1984,
109 pp.
11. Water Quality Management in Aquaculture. CMFRI
Special Publication No. 22, 1985, 96 pp.

* Out of print.

•
 -.3
FERTILIZATION

3.1. Chemical fertilizers

Inorganic fertilizers used in ponds are the same ones used for
agricultural crops. Nitrogen, phosphorus and potassium are
termed the primary nutrients in fertilizers. The grade of a
fertilizer refers to percentages by weight of nitrogen (as N),
phosphorus (as P2Os) and potassium (as K,0, also called potash).
For example, a 20-20-5 grade fertilizer contains 20% N,
20%P,O6 and 5% KaO. This method of expressing nitrogen,
phosphorus and potassium content is traditional rather than
descriptive. Fertilizer does not contain elemental nitrogen (N).
phosphorus pentoxide (P«05) or potash (KaO). The use of N,
Pa06 and KaO to indicate fertilizer grades originated long ago
and has been accepted for practical purpose. Primary nutrients
in fertilizers are usually present as relatively simple compounds
which dissolve to give nitrate, ammonium, phosphate or
potassium ions. The compositions of some common fertilizer
materials are given in Table 3.1. Calcium, magnesium and

TABLE 3.1. Composition of some common inorganic fertilizer

materials

Material Content (%)

N P,0, K,0
Ammonium nitrate .. 33-35 —_ —
Ammonium sulphate '.. 20-21 — —
Calcium metaphosphate — 62-64 —
Calcium nitrate .. 15.5 — —
Ammonium phosphate .. 11-16 20-48 —
Muriate of potash — -ri 5042
Potassium nitrate .. 13 •— . . . 4 4 - ;
Potassium sulphate — — 50
Sodium nitrate .. 16 — • —

Superphosphate (ordinary) . . • — 18-20 —

Superphosphate (double or triple) — 32-54 —