SOURCES OF ENERGY FOR FISH AND CALCULATING

ENERGY DIGESTIBITY

NUTRITIONAL BIOENERGETICS IN FISH

ENERGY METABOLISM IN FISH

ENERGY SOURCES

ENERGY REQUIREMENTS OF FISH

ENERGY METABOLISM IN FISH

1. Energy Flow in Animals

2. Energy Loss

Energy is needed for the maintenance of all living organisms. Most plants obtain their energy

directly from the sun and use that energy to synthesize the complex molecules which make up the

structural and storage parts of the plant. Animals cannot utilize/radiant energy from the sun. They

get their needed energy from oxidation of the complex molecules which are eaten by the animal.

The energy in feed is not available until the complex molecules are broken down to simpler

molecules by digestion. The products of digestion are then absorbed into the body of the animal

where oxidation processes occur which release the energy. Energy metabolism in fish is similar

to that in mammals and birds with two notable exceptions. These exceptions are:

(a) fish do not expend energy to maintain a body temperature different from that of their

environment; and

(b) the excretion of waste nitrogen requires less energy in fish than it does in homeothermic land

animals.

There are large differences in the ability of different species of fish to digest feed materials. Fish

species range all the way from strict herbivores through omnivores to carnivores. The food

requirements of different species of fish vary greatly. The job of the nutritionist is to identify the

needs of the animal and then to find feed materials which will most economically satisfy these

requirements.

1. Energy Flow in Animals

It should be kept in mind that energy needs for maintenance and voluntary activity must be

satisfied before energy is available for growth. Also during times of low food intake fat and protein

are withdrawn from the animal body to provide the energy needs for maintenance and the animal

loses weight.

2. Energy Loss
Energy is lost from the body of a fish in the faeces, urine, gill excretions and as heat. Also small

amounts are lost from external body surface.

The energy lost as heat comes from three sources which are difficult to measure separately. These

are:

(a) standard metabolism (SM), which is the energy required to keep the animal alive and is similar

to basal metabolism measured in humans. Because of the difficulty of obtaining a "motionless"

animal, the definition of basal metabolism is not applicable to fish. When a fish is restrained to a

motionless condition it struggles to free itself and uses more energy than if allowed to swim freely

in still water. SM is the minimum heat production of an undisturbed fish in the "post absorptive"

state in still water;

(b) voluntary physical activity, which is the energy expended by a fish moving about, seeking

food, maintaining position, etc.;

(c) heat of nutrient metabolism, also called heat increment or specific dynamic action (SDA),

which is the heat released by the many chemical reactions associated with the processing of

ingested feed. It includes the energy expended in digestion, absorption, transportation, and

anabolic activities. It also includes the cost of excretion of waste products.

ENERGY SOURCES

1. Fats

2. Carbohydrates

3. Protein

Energy is stored in the chemical structure of the complex molecules of feed materials. When

oxidation occurs, energy is released and is available to do work. This released energy is trapped

by biochemical reactions and is used to drive the energy requiring reactions necessary to sustain

life.

The energy needs of fish are supplied by fats, carbohydrates, and proteins.

1. Fats
Fats are the principal form of energy storage in plants and in animals. Fat contains more energy

per unit weight than any other biological product. The inclusion of fat usually increases the

palatability of a feed. Generally fats are well digested and utilized by fish. There is little hard data

on the ability of fish to digest fats of different melting points. It is usually estimated that fat

provides 8.5 kcal metabolizable energy (ME) per gramme. The fatty acid products of digestion

are well utilized by most fish. There is some evidence that high levels of short chain fatty acids

can depress growth. This is seldom a problem in practical diets.

Natural diets may contain as much as 50 percent fat. High levels of fat can also be used in

manufactured feeds if other nutrients are adequate. Full fat oilseed meals may be the most practical

way to add fats.

2. Carbohydrates

Carbohydrates are the cheapest and most abundant source of energy for animals. Most of plant

material is carbohydrate. Carbohydrates in feed material range from easily digested sugars to

complex cellulose molecules which cannot be digested by animals. It is only through their

symbiotic relationship with bacteria that ruminant animals can utilize large amounts of cellulose.

There is controversy as to the value of carbohydrate in fish feeds. It appears, however, that

digestible carbohydrate can be well utilized as an energy source if it is kept in proper balance with

other nutrients.

The ME values of carbohydrates for fish range from near zero for cellulose to about 3.8 kcal/g for

easily digested sugars. Raw starch ranges from 1.2 to 2.0 kcal ME/g. Cooking of starch can

increase the ME to about 3.2 kcal/g. Heat and moisture associated with the pelleting process

improves the digestibility of starchy feed materials. The value of carbohydrate in fish diets

depends on the source and type of carbohydrate and the processing to which it has been subjected.

3. Protein

In nature, carnivorous fish consume diets which are about 50 percent protein. Fish have a very

efficient system for excretion of waste nitrogen from protein which is catabolized for energy and
therefore high protein diets are not harmful. Protein is often the most expensive source of energy

in manufactured diets and should be kept to a minimum, consistent with good growth and feed

conversion. Protein has a ME value of about 4.5 kcal/g for fish, which is higher than that for

mammals and birds. The low energy cost of excreting waste nitrogen in fish is primarily

responsible for this.

In general, proteins from animal sources are more digestible than those from plant sources.

Processing methods can also influence protein quality. Heating increases the digestibility of some

proteins and reduces that of others. Protein is used very efficiently by fish as a source of energy

but for economic reasons should be kept to a minimum, consistent with good growth, and cheaper

carbohydrate and fat should be used to supply most of the energy.

ENERGY REQUIREMENTS OF FISH

1. Energy Distribution in Relation to Feeding Level

2. Maintenance Energy

3. Energy Cost of Growth

4. Factors that Alter Energy Needs

As discussed earlier the energy needs for maintenance and activity must be satisfied before any

growth can occur. Feeding levels must be high enough to supply maintenance needs and still have

energy remaining for growth. Digestion efficiency in fish decreases as feeding level is increased.

The problem becomes one of finding the feeding level at which the increased efficiency of energy

utilization at a high feeding rate is balanced by the lower efficiency of digestion at the higher

feeding rate

1. Energy Distribution in Relation to Feeding Level

It has been studied the distribution of dietary energy intake in relation to feeding level in fish.

Basal or standard metabolism in fish is relatively constant under constant environmental

conditions. It can change with changes in temperature and fish size among other factors. The

energy expended on voluntary activity usually increases somewhat with increasing feeding level.

Starved fish are less active than well-fed fish but there is always some expenditure of energy for
activity. The heat of nutrient metabolism is proportional to the level of feeding. The energy

excreted in urine and gill excretions is also a function of feeding level. The reduced efficiency at

high levels of feeding is uniformly large area representing faeces at high levels of feeding. The

amount remaining for growth is zero at maintenance feeding and becomes proportionately greater

as feeding level is increased, until it is balanced by the decreased efficiency, of digestion.

2. Maintenance Energy

All of the energy lost due to standard metabolism, heat of nutrient metabolism and physical

activity appears as heat. The maintenance requirement can be determined by measuring the heat

produced. The heat production can be measured directly in a calorimeter or it can be estimated by

measuring oxygen and applying the appropriate heat equivalent. The factor most commonly used

is 3.42 kcal/mg O2. This factor is largely an extrapolation of data for mammals and has not been

directly measured in fish. The heat equivalent of oxygen also varies with the type of substrate

being oxidized. Maintenance energy can also be estimated by measuring energy loss during

starvation.

3. Energy Cost of Growth

It has been shown in mammals that the cost of growth is fairly constant after maintenance energy

is subtracted from ME fed. This probably holds true for fish, but it has not been experimentally

determined. More research is needed in this area.

4. Factors that Alter Energy Needs

There are several factors which can alter the energy requirements of fish. Feeding rates should be

adjusted to compensate for these factors to avoid overfeeding, but still providing sufficient energy

for optimum growth.

(a) Temperature: As environmental temperature declines homeotherms must increase their

metabolic rate to compensate for the additional heat loss if they are to maintain a constant body

temperature. Most freshwater fish do not attempt to maintain a body temperature which is different

from the environment. As water temperature declines, body temperature of the fish declines and

metabolic rate is reduced. The low metabolic rate at low temperatures enables fish to survive for
long periods under ice where little food is available. There is considerable species difference in

metabolic adaptation to environmental temperature changes. Each species seems to have a

preferred temperature at which it functions most efficiently. If temperature gradients exist, the

fish will seek the most favourable temperature. Usually this is the temperature at which the

difference between maintenance requirement and voluntary food intake is greatest and at which

optimum efficiency of growth occurs.

(b) Water Flow: Energy which is used for physical activity is not available for growth. Fish which

are forced to swim against a strong current are expending energy which would otherwise be used

for growth. However, still water allows stratification and the accumulation of waste products. Fish

rearing facilities should be designed to obtain maximum use of water without undue stress on the

fish.

(c) Level of Feeding. The level of feeding also has an effect on the energy expenditure of fish.

This becomes important in design of fish rearing facilities. Dissolved oxygen is usually the first

limiting factor in fish rearing. The oxygen consumption increases shortly after feeding due to the

physical activity of feeding and the heat of nutrient metabolism. Facilities must be designed with

adequate safety margins. The oxygen required per unit weight of feed also varies with feeding

level, being higher at maintenance level when all the food is oxidized than at higher feeding levels

when much of the energy is stored as growth.

(d) Other Factors: Several other factors can contribute to high energy requirements. Anything

which makes the fish uncomfortable increases physical activity and reduces growth. Crowding,

low oxygen and waste accumulation are some of these factors.

PROTEINS AND AMINO ACIDS

GROSS PROTEIN REQUIREMENTS

Gross protein requirements have been determined for a few species of fish. Simulated whole egg

protein component of test diets contains an excess of indispensable amino acids. These diets were

kept approximately isocaloric by adjusting total protein plus digestible carbohydrate components

to a fixed amount as the protein diet treatments were varied over the ranges tested. Tests in feeding
fry, fingerling, and yearling fish have shown that gross protein requirements are highest in initial

feeding fry and that they decrease as fish size increases. To grow at the maximum rate, fry must

have a diet in which nearly half of the digestible ingredients consist of balanced protein; at 6-8

weeks this requirement is decreased to about 40 percent of the diet for salmon and trout and to

about 35 percent of the diet for yearling salmonids raised at standard environmental temperature

(SET). Gross protein requirements for young Catfish appear to be less than those for salmonids.

Initially feeding fry require that about 50 percent of the digestible components of the ration be

protein, and the requirement decreases with size. Some feeding trials with salmon have indicated

direct relationships between changes in the protein requirements of young fish and changes in

water temperature. Chinook salmon in 7 C water require about 40 percent whole egg protein for

maximum growth; the same fish in 15 C water require about 50 percent protein. Salmon, trout and

catfish can use more protein than required for maximum growth because of efficiency in

eliminating nitrogenous wastes in the form of soluble ammonia compounds through the gill tissue

directly into the water environment. This system for eliminating nitrogen is more efficient than

that available to fowl and mammals. Fowl and mammals consume energy to synthesize urea, uric

acid, or other nitrogen compounds which are excreted through the kidney tissue and expelled in

urine. Digestible carbohydrate and fat will spare excess protein in the diet as long as the protein

requirement for maximum growth is met.

Basically the fish must be given a diet containing graded levels of high quality protein and energy

and adequate balances of essential fatty acids, vitamins and minerals over a prolonged period.

From the resulting dose/response curve the protein requirement is usually obtained by an Almquist

plot. These differences in apparent protein requirement are thought to be due to differences in

culture techniques and diet composition.

The relatively high dietary protein levels required for maximal growth of certain fish such as grass

carp, Ctenopharyngodon idella, and Brycon spp. are surprising as these fish are omnivorous.

Brycon spp. are grown on unwanted fruit and other plant material of low protein content and under
these conditions there is presumably a substantial contribution to their protein intake from a

natural food chain.

Protein requirement of eurythaline fish such as the rainbow trout, Salmo gairdneri, and the coho

salmon, Oncorhynchus kisutch, reared in water of salinity 20 ppt is about the same as the

requirement in freshwater. No data are available for the protein requirement of these species in

full strength sea water.(35 ppt).

AMINO ACIDS

The amino acids are the building blocks of proteins; about 23 amino acids have been isolated from

natural proteins. Ten of these are indispensable for fish. The animal is incapable of synthesizing

indispensable amino acids and must therefore obtain these from the diet.

Essential and Non-essential Amino Acids

Salmon, trout and channel catfish fed diets devoid of arginine, histidine, isoleucine, leucine,

lysine, methionine, phenylalanine, threonine, tryptophan or valine failed to grow (Fig. 3). These

same fish fed diets devoid of other L-amino acids grew as well as fish receiving all 18 amino acids

tested. The nitrogen component in the test diets was made up of 18 L-amino acids in the pattern

found in whole egg protein. All fish on test recovered rapidly when the missing amino acid was

replaced in the diet. The slope of the growth curve of the recovery group was identical with that

of fish receiving the complete amino acid test diet.

Dispensable amino acids tested were alanine, aspartic acid, cystine, glutamic acid, glycine,

proline, serine, and tyrosine. These amino acids were found to be not essential for the growth of

salmon, trout and channel catfish.

Quantitative studies on the requirements of the 10 indispensable amino acids used a casein-gelatin

mixture supplemented with crystalline L-amino acids. The test diet had an amino acid pattern of

40 percent whole egg protein for the nitrogen component. Experiments conducted with carp and

eel showed a similar lack of growth when an indispensable amino acid was absent from the diet.

QUANTITATIVE REQUIREMENTS OF AMINO ACID

Quantitative requirements by salmonids for the ten indispensable amino acids were determined

by feeding linear increments of one amino acid at a time in a test diet containing an amino acid

profile identical with whole egg protein except for the amino acid tested. Replicate groups of fish

were fed the diet treatments until gross differences appeared in the growth of test lots. An

Almquist plot of growth response indicated the level of amino acids required for maximum growth

under those specific test conditions. Diets were designed to contain protein at or slightly below

the optimum protein requirement for that species and test condition to assure maximum utilization

of the limiting amino acid. A recent innovation has been the use in test diets of proteins relatively

deficient in a given essential amino acid. Thus combinations of fishmeal and zein have been used

in test diets to define the requirement of rainbow trout for arginine. Diets containing different

relative amounts of casein and gelatin showed that an increase in the level of protein-bound

arginine from 11 to 17 g/kg resulted in a significant increase in the growth of channel catfish.

Arginine requirement of rainbow trout has been determined from a conventional dose/response

(growth) curve and also by measuring the tissue (blood and muscle) levels of free arginine in

groups of trout given increasing amounts of dietary arginine. After the dietary requirement of the

trout for arginine has been met, any further increase in arginine intake led to an increase in the

concentration of free arginine in blood and muscle. Good agreement was obtained between the

two methods. The data suggested that real differences exist between fish species in their

requirement for certain amino acids. This leads to difficulties in formulating the protein

component of practical diets for those species whose amino acid requirements are not yet known.

A possible solution is to use, for each amino acid, the highest level required by any of those species

for which data is available. The need for further quantitative data on the amino acid requirements

of fish, especially those actually or potentially useful as farm animals, is obvious.

SUPPLEMENTING DIETS WITH AMINO ACIDS

One solution to the use of proteins that are relatively deficient in one or more amino acids is to

supplement the protein with appropriate amounts of the amino acid needed in practical diets. Fish

appear to utilize free amino acids at various degrees of efficiency.

Young carp, Cyprinus carpio, were shown to be unable to grow on diets in which the protein

component (casein, gelatin) was replaced by a mixture of amino acids similar in overall

composition. A trypsin hydrolyzate of casein was equally ineffective. However, if a diet

containing free amino acids as the protein component is carefully neutralized with NaOH to pH

6.5-6.7 then some growth of young carp does occur. This growth was markedly inferior to that

occurring on a comparable casein diet under the same conditions. Channel catfish are also unable

to utilize free amino acids given as supplements to deficient proteins. When soybean meal was

substituted isonitrogenously for menhaden meal, growth and feed efficiency of channel catfish

were substantially reduced. Addition of free methionine, cystine or lysine, the most limiting amino

acids, to these soy-substituted diets did not enhance weight gain. Raising the arginine level of

catfish diets from 11 to 17 g/kg by isonitrogenous substitution of gelatin for casein enhanced

weight gain significantly but the addition of free arginine, cystine, tryptophan or methionine to

casein had little effect on growth or food conversion. Salmonids are able to utilize free amino

acids for growth. A zein-gelatin diet supplemented with lysine and trytophan was shown to be

markedly superior to an unsupplemented zein-gelatin diet for rainbow trout when weight gain and

protein utilization were used as criteria. Several investigators have demonstrated the potential of

supplementing amino acid deficient proteins with limiting amino acids in diets for salmonids.

Casein supplemented with six amino acids produced feed conversion ratios with Atlantic salmon

similar to those obtained when an isolated fish protein was used as the dietary protein source.

Soybean meal supplemented with five or more amino acids (including methionine and lysine) was

a superior protein source to soybean meal alone for rainbow trout. Single additions of methionine

and lysine did not, however, improve the value of soybean meal. These results suggest that the

amino acid spectrum of the isolated fish protein they used may possibly approximate the amino

acid requirement of rainbow trout. The nutritional value of a soy protein isolate could be enhanced

by supplementing it with the first limiting amino acid; i.e., methionine. Diets containing, as

protein component, fishmeal, meat and bone meal, and yeast and soybean meal could be improved

by supplementing with cystine (10 g/kg) and tryptophan (5 g/kg) together. Fishmeal can be
entirely replaced without a reduction in food conversion rate in diets for rainbow trout by a

mixture of poultry by-product meal and feather meal together with 17 g lysine HCL/kg, 4.8 g DL-

methionine/kg, and 1.44 g DL-tryptophan/kg.

LIPIDS AND FATTY ACIDS

Lipids are the generic names assigned to a group of fat soluble compounds found in the tissues of

plants and animals,: and are broadly classified as:

a) fats, b) phospholipids, c) sphingomyelins, d) waxes, and e) sterols.

Fats are the fatty acid esters of glycerol and are the primary energy depots of animals. These are

used for long-term energy requirements during periods of extensive exercise or during periods of

inadequate food and energy intake. Fish have the unique capability of metabolizing these

compounds readily and, as a result, can exist for long periods of time under conditions of food

deprivation. A typical example is the many weeks of migration by salmon in their return upstream

to spawn; stored lipid deposits are burned for fuel to enable body processes to continue during the

strenuous journey.

Phospholipids are the esters of fatty acids and phosphatidic acid. These are the main constituent

lipids of cellular membranes allowing the membrane surfaces to be hydrophobic or hydrophylic

depending on the orientation of the lipid compounds into the intra or extracellular spaces.

Sphingomyelins are the fatty acid esters of sphingosine and are present in brain and nerve tissue

compounds. Waxes are fatty acid esters of long-chain alcohols. These compounds can be

metabolized for energy and to impart physical and chemical characteristics through the stored

lipids of some plant and several animal compounds.

Sterols are polycyclic, long-chain alcohols and function as components of several hormone

systems, especially in sexual maturation and sex-related physiological functions.

Fatty acids can exist as straight chain or branch chain components; many of the fish fats contain

numerous unsaturated double bonds in the fatty acid structures. A short bond designation for. fatty

acids will be used throughout where the w number identifies the position of the first double bond

counting from the methyl end. Linolenic acid would be written 18:3w 3. The first number
identifies the number of carbons; the second number, the number of double bonds; and the last

number, the position of the double bonds.

Many reviews of fish nutrition have been published which contain information on lipid

requirements. Most work on lipid requirements of fish has been with salmonids. Rainbow trout

have an essential fatty acid (EFA) requirement for the linolenic of w 31 series rather than for

linolenic or w 6 as required by most mammals. The main emphasis on lipid requirements has been

on EFA and on the energy value of lipids.

CARBOHYDRATES

Carbohydrates represent a broad group of substances which include the sugars, starches, gums

and celluloses. The common attributes of carbohydrates are that they contain only the elements

carbon, hydrogen and oxygen, and that their combustion will yield carbon dioxide plus one or

more molecules of Water.

The simplest carbohydrates are the three-carbon sugars which figure importantly in intermediary

metabolism and the most complex are the naturally occurring polysaccharides, primarily of plant,

origin. In the diet of animals and fish, two classes of polysaccharides are significant:

(a) structural polysaccharides which are digestible by herbivorous species -cellulose, lignin,

dextrans, mannans, inulin, pentosans, pectic acids, algic acids, agar and chitin; and

(b) universally digestible polysaccharides - principally starch.

Carbohydrates make up three-fourths of the biomass of plants but are present only in small

quantities in the animal body as glycogen, sugars and their derivatives. Glycogen is often referred

to as animal starch because it is not present in plants. Derived mono-saccharides such as the sugar

acids, amino sugars and the deoxysugars are constituents of all living organisms.

Much of the carbohydrates that enter the diets of animals, including fish, is of plant origin.

Carnivorous fish like the Atlantic salmon and the Japanese yellowtail, therefore, deal with little

carbohydrate,. Indeed, experiments have shown that these species are ill-equipped to handle

significant quantities of raw carbohydrate, in their diets. On the other hand, omnivores such as the
common carp and the channel catfish are able to digest fair amounts of carbohydrates in their

diets. The grass carp, a herbivore, subsists primarily on a vegetarian diet.

THE VITAMINS

1. WATER-SOLUBLE VITAMINS

2. FAT-SOLUBLE VITAMINS

1. WATER-SOLUBLE VITAMINS

1.Thiamine

2. Riboflavin

3. Pyridoxine

4. Pantothenic Acid

5. Niacin

6. Biotin

7. Folic Acid (Folacin)

8. Vitamin B12

9. Ascorbic Acid

10. Inositol

11. Choline

12. p-Aminobenzoic Acid

13. Lipoic Acid

Water-soluble vitamins include eight well-recognized members of the vitamin B complex:

thiamine, riboflavin, pyridoxine, pantothenic acid, niacin, biotin, folic acid and vitamin B12; the

water-soluble essential nutritional factors: choline, inositol, ascorbic acid; and vitamins with less-

defined activity for fish: p-aminobenzoic acid, lipoic acid and citrin. The first eight, though

required only in small amounts in the diet, play major roles in growth, physiology and metabolism.

Choline, inositol and ascorbic acid are required in appreciable quantities in the diet and sometimes

are not referred to as vitamins but as major dietary nutrients.

THE MINERALS

Mineral elements have a great diversity of uses within the animal body. The following mineral

elements are recognized as essential for body functions in fish: calcium, phosphorus, sodium,

molybdenum, chlorine, magnesium, iron, selenium, iodine, manganese, copper, cobalt and zinc.

To these may be added fluorine and chromium which have also been shown to be essential for

land animals. The prominence of each mineral element in body tissues is closely related to its

functional role. As constituents of bones and teeth, minerals provide strength and rigidity to

skeletal structures. In their ionic states in body fluids they are indispensable for the maintenance

of acid-base equilibrium and osmotic relationship with the aquatic environment, and for

integration activities involving the nervous and endocrine systems. As components of blood

pigments, enzymes and organic compounds in tissues and organs they are indispensable for

essential metabolic processes involving gas exchange and energy transactions.

FISH FEED FORMULATION

• INTRODUCTION

• BALANCING CRUDE PROTEIN LEVEL

• STEPS IN FEED FORMULATION

• BEST-BUY TECHNIQUES

INTRODUCTION

Feed formulation is essentially applied nutrition. A number of terms and expressions are

introduced that will be put to practical use as information is presented on the nature and qualities

of various feedstuffs and the information presented on the nutrient requirements of fish. Precise

understanding of these terms is essential to their correct application. One must recognize that some

of these terms have a built-in error that cannot be escaped. This does not eliminate their usefulness

in feed formulation. However, one must appreciate the fact that some are useful approximations

of the values and not true values. The terms that one needs to understand to formulate practical

fish diets are: crude protein level; energy level, either expressed as metabolizable energy (ME) or
as digestible energy (DE); specific amino acid levels; crude fibre level; and ash level. Since most

complete practical fish diets are supplemented with a vitamin premix at levels in excess of the

dietary requirement, this category of nutrients will be ignored temporarily. The potential problems

occur when one fails to recognize that all of the above mentioned terms, except ME and DE,

represent the quantity or level of a nutrient in the feed as determined by chemical tests on a specific

sample of a feedstuff. These chemical tests generally correlate well enough with biological

methods of feed evaluation (growth studies, tissue, levels) to be very useful to feed formulators,

but they are still chemical tests that are subject to experimental error during nutrient level

determination. For example, the proximate composition of fish meals changes during the

spawning season. Generally, the lipid levels increase before spawning and decrease after

spawning. This will alter the percent of protein, ash, and carbohydrates in fish meal as the seasons

change. Similarly, many plant feedstuffs vary in proximate composition with their stage of

maturity at harvest, location grown, and other environmental conditions, such as the weather.

Tabled values represent an average value that is usually close enough to the actual value to allow

accurate feed formulation. However, one must be aware that assumptions are being made in order

to recognize the potential sources of error that may exist.

Metabolizable, energy and digestible energy values are obtained biologically and, thus, should

accurately represent the true energy value of feedstuffs to fish. However, ME values may be

obtained in different ways (faeces collection methods) and thus may be subject to experimental

error. It has recently been reported that the digestibility of feed by rainbow trout was lower at 7°C

than at 11°C or 15°C. At 11°C and 15°C body size (18.6 g, 207.1 g or 585.7 g) did not affect feed

digestibility. The digestibility of carbohydrate and energy was slightly reduced by meal size in

rainbow trout fed at 1.6 percent body weight. Protein and lipid digestibility was not reduced by

meal size. Obvious differences exist between fish species in nutrient digestibility, especially in

the carbohydrate fraction of feed. Herbivorous and, to a lesser extent, omnivorous fish have longer

digestive tracts than do carnivorous fish and are able to obtain more digestible energy from

carbohydrates. An awareness of these facts will prevent misuse of ME and DE values.

Each feedstuff in any diet formulation should be present for a specific reason; i.e., it is a good

energy source, it is rich in a limiting amino acid, etc. In addition, each feedstuff in a particular

diet formulation should be the least costly ingredient available for its particular function in the

diet. This leads to another assumption in feed formulation; that is, any nutrient in a particular

feedstuff, such as an amino acid, is just as valuable as the same nutrient in any other feedstuff.

This allows feed formulators to interchange one feedstuff with another as cost and availability

change. Thus, it is assumed that there is no "ideal formulation", but rather an almost infinite

number of possible feed formulations that met the nutritional needs of the fish equally well. While

this assumption may not be entirely valid and some nutritional judgement must be employed in

any feed formulation, it does seem to be valid in most cases. As with the previously mentioned

assumption, an awareness of the potential pitfalls involved is necessary for the fish feed

formulation so that allowances can be made in diet formulation and problems can be anticipated

and avoided.

BALANCING CRUDE PROTEIN LEVEL

In most animal diets, protein is the most expensive portion and is usually the first nutrient that is

computed in diet formulation. The energy level of the diet is then adjusted to the desired level by

addition of high energy supplements) which are less expensive than protein supplements. The

square method is an easy way to determine the proper dietary proportions of high and low protein

feedstuffs to add to a feed to meet the dietary requirement of the animal to be fed.

For example, suppose rice bran and soybean meal were available as feedstuffs to prepare a diet

for carp that was 25 percent crude protein. A square is constructed and the two feedstuffs are put

on the two left corners along with the protein content of each. The desired protein level of the feed

is placed in the middle of the square. Next, the protein level of the feed is subtracted from that of

the feedstuffs, placing the answer in the opposite corner from the feedstuff. Ignore positive or

negative signs.

If more than two feedstuffs are used in a feed, they may be grouped into basal feeds (CP < 20

percent) and protein supplements (CP > 20 percent), averaged within each group, and plugged
into the square method. For example, suppose shrimp meal and corn were also available for the

carp feed mentioned above. The crude protein levels of the shrimp meal (52.7 percent) and of corn

(10.2 percent) are averaged with soybean meal and rice bran, respectively.

The square method is helpful to novice feed formulators because it can get them started in diet

formulation without the need to resort to trial and error. The square method can also be used to

calculate the proportion of feeds tuffs to mix together to achieve a desired dietary energy level as

well as a crude protein level. If one wanted to make a feed containing 2 500 kcal ME/kg using

wheat middlings (1663 kcal ME/kg) and anchovy fish meal (4 371 kcal, ME/kg) a square could

be constructed as follows:

STEPS IN FEED FORMULATION

The first step in diet formulation is balancing the crude protein and energy levels. This can be

accomplished by trial and error, by the square method for either crude protein level or energy level

and then adjusting, or by solving simultaneous equations. At first, it is helpful to use at least three

feeds tuffs during the initial balancing of protein and energy levels: one high in protein and high

in ME, one low or intermediate in protein and high in ME, and one low or intermediate in both

protein and ME. Once practice makes one more proficient at diet formulation any number of

feedstuffs can be used. One must remember to reserve room in the formulation for any feed

additive, such as a vitamin or mineral pre-mix.

The second step in diet formulation is to check the levels of indispensable amino acids in the

formulation to be sure the dietary levels meet the requirements of the animal to be fed. The

requirements of fish for indispensable amino acids is expressed as the dietary level (as a percent

of the diet) or as a percent of the dietary protein level. To convert an amino acid level from the

percent of diet to percent of protein, divide the dietary level of each amino acid by the dietary

protein level. It might be of interest to calculate the dietary levels of all of the indispensable amino

acids, but it is not practical to do it all of the time. If the levels of arginine, lysine, methionine,

and tryptophan meet the dietary requirements of the fish to be fed, the levels of the other six
indispensable amino acids will most likely be above required levels. When using unconventional

protein supplements, the levels of all ten indispensable amino acids should be checked.

If the diet formulation is low in any amino acid, a feedstuff that contains high levels of that amino

acid must be added to the diet at the expense of another ingredient. Once the amino acid

requirements are met, the dietary protein and energy levels must be rechecked to, see if any

substitution of ingredients has imbalanced the formulation. The amino acids listed are for

illustration purposes only and may be changed to suit different circumstances. In practical feed

formulation, pellet quality and acceptability must be considered in addition to nutrient levels and

cost. These considerations will vary from species to species and with the type of pellet being

made, and are dealt with in other sections of this manual.

BEST-BUY TECHNIQUES

The price of the feedstuffs used in diet formulations must be considered to formulate a cost-

efficient diet. Feedstuffs can be compared with one another on the basis cost per unit of protein,

energy, or amino acid. For example, suppose one has wheat middlings and wheat millrun available

for a fish diet, which feedstuff would be the least expensive source of energy?

The cost of protein is often the greatest part of the cost of a fish diet. Therefore, substantial savings

can be made by using best-buy techniques to determinate least expensive protein supplement. To

compare anchovy meal and herring meal, the following calculations are made:

To compare feedstuffs on the basis of cost per unit of an amino acid, one can calculate the best

buy in the same way as before. For example, sesame oil cake which has twice as much methionine

content as does groundnut cake on a per unit protein basis would be a more attractive buy at

comparable prices. These kinds of comparisons are only valid if the nutrient in one feedstuff is as

valuable or available to the animal as the same nutrient in another feed. Such comparisons should

be made whenever prices change.

REFERENCES
Ackman, R.G., 1967 Characteristics of the fatty acid composition and biochemistry of some

freshwater fish oils and lipids in comparison with marine oils and lipids.

Comp.Biochem.Physiol., 22:907-22

Brody, S., 1945 Bioenergetics and growth. London, Hafner Press

Castell, J.D., 1979 Review of lipid requirements of finfish. In Finfish nutrition and fishfeed

technology, edited by J.E. Halver and K. Trews. Proceedings of a World Symposium

sponsored and supported by EIFAC/FAO/ICES/IUNS, Hamburg, 20-23 June, 1978.

Schr.Bundesforschungsanst.Fisch.,Hamb., (14/15) vol.1: 59-84

Cowey, C.B. and J.R. Sargent, 1972 Fish nutrition. Adv.Mar.Biol., 10:383-492

Cowey, C.B. and J.R. Sargent, 1972 Fish nutrition. Adv.Mar.Biol., 10:383-492,

Cowey, C.B. and J.R. Sargent, 1977 Lipid nutrition in fish. Comp.Biochem.Physiol. (B

Comp.Biochem.) 57:269-73

Cowey, C.B., 1979 Protein and amino acid requirements of finfish. In Finfish nutrition and

fishfeed technology, edited by J.E. Halver and K. Tiews. Proceedings of a World

Symposium sponsored by EIFAC/FAO, ICES and IUNS, Hamburg, 20-23 June, 1978.

Schr.Bundesforschungsanst.Fisch.,Hamb., (14/15)vol. 1:3-16

Harder, W. 1975, Anatomy of fishes. Part I. Text. Part 2. Figures and plates. Stuttgart. E.

Schweizerbart'sche Verlagsbuchhandlung, Pt.1:612 p., Pt.2:132 p. 13 pl.

Kapoor, B.B. 1975, H. Smit and I.A. Verighina, The alimentary canal and digestion in teleosts.

Adv.Mar.Biol. 13:109-239

Kleiber, M., 1961 The fire of life. New York, Robert E. Krieger Publishing Co.

Magnuson, J.J. 1969, Digestion and food consumption by Skipjack tuna. Trans.Am.Fish.Soc.,

98(3): 379-92

Maynard, L.A. and J.K. Loosli, 1962 Animal nutrition. New York, McGraw-Hill Book Co

McLaren, B.A. 1947 et al., The nutrition of rainbow trout. 1. Studies of vitamin requirements.

Arch-Biochem. 19:169-78
Meed, J.F. and M. Kayama, 1967 Lipid metabolism in fish. In Fish oils, edited by M.E. Stansby.

Westport, Conn., Avi Publ. Co., pp. 289-99

Mertz, E.T., 1972 The protein and amino acid needs. In Fish nutrition, edited by J.E. Halver. New

York, Academic Press, pp. 106-43

National Research Council, 1973 Subcommittee on Fish Nutrition, Nutrient requirements of trout,

salmon and catfish. Washington, D.C., National Academy of Sciences, (Nutrient

requirements of domestic animals), 11:57 p.

National Research Council, 1977 Subcommittee on Warmwater Fishes, Nutrient requirements of

warmwater fishes. Washington, D.C., National Academy of Sciences (Nutrient

requirements of domestic animals) 78 p.

National Research Council, 1977 Subcommittee on Warmwater Fishes, Nutrient requirements of

warmwater fishes. Washington, D.C., National Academy of Sciences, (Nutrient

requirements of domestic animals), 78 p.

National Research Council, 1977 Subcommittee on Warmwater Fishes, Nutrient requirements of

warmwater fishes. Washington, D.C., National Academy of Sciences, (Nutrient

requirements of domestic animals) 78 p.

Phillips, A.M. Jr., 1969 Nutrition, digestion and energy utilization. In Fish physiology, edited by

W.S. Hoar and D.G. Randall. New York, Academic Press, vol. 1:391-432

Post, G., W.E. Shanks and R.R. Smith, 1965 A method for collecting metabolic excretions from

fish. Prog.Fish-Cult. 27:108-88

Prosser, C.L. (ed.),1973 Comparative animal physiology. Philadelphia, W.B. Saunders Company,

1011 p. 3rd ed.

Sinnhuber, R.O., 1969 The role of fats. In Fish in research, edited by O.W. Newhaus and J.E.

Halver, New York, Academic Press, pp. 245-61

Smirnova, L.I., 1966 Digestive leukocytosis of bream (Abramis brama). In Biology of fishes of

the Volga reservoirs. Tr.Inst.Biol.Vnutr.Vod./Trans.Inst.Biol.Inland Waters, 10(13) -.143-

7
Smith, B.W. and R.T. Lovell, 1973 Determination of apparent protein digestibility in feeds for

channel catfish. Trans.Am.Fish.Soc., 102(4):831-5

White, A., et al., 1978 Principles of biochemistry. New York, McGraw-Hill Book Company, 1492

p. 6th ed.

Winberg, G.G., 1956 Rate of metabolism and food requirements of fishes. Transl.Serv.Fish.Res

Board Can., (194)

Yoshinaka, R., M. Sato and S. Ideka, 1973 Studies on collagenase of fish. l. Existence of colla-

genolytic enzyme in pyloric caecae of Seriola quinqueradiata. Bull. Japan. Soc. Sci.Fish.,

39(3):275-81
A-Multiple choice questions

1. The gut is a tubular structure beginning at the mouth and ending at

……………………………

a) Anus b) mouth c) lungs d) tail

2. The gut wall in lampreys also contains …………………..fibers.

a) diagonal muscle b) circular muscle c) Longitudinal muscle d) none of these

3. Generally, …………… guts are found in herbivore fishes.

a) smaller b) longest c) curvy d) no gut

4. The mouth exhibits a variety of fascinating adaptations for capturing, holding and sorting

food, ratcheting it into the …………………..

b) Esophagus b) stomach c) intestine d) none of these

5. The common carp provides an excellent example of ……………………………being

used as the primary chewing apparatus.

a) Milk teeth b) non-mandibular teeth c) mandibular teeth d) permanent teeth

6. ………………………….consists of a travelling wave of contraction of the circular and

longitudinal layers of muscle in the gut wall such that material inside the gut is moved

along.

a) Diffusion b) osmosis c) Peristalsis d) active transport

7. …………………..is the predominant gastric enzyme of all vertebrates, including fish.

a). Pepsin b) amylase c) both d) none of these

8. Tryptic activity has been found in four stomach-less species in Japan: Seriola, two basses

and a puffer.

a) Seriola b) tune c) shark d) hag

9. …………………………..are largely carnivorous, but show few anatomical

specializations for capturing and digesting prey.

a) Common carp b) Rainbow trout c) Milkfish d) none

10. Carp lack a …………………, but have a long intestine which winds extensively

throughout the visceral cavity.

a) Intestine b) pancreas c) stomach d) mouth

11. ………………. are most frequently designated as microphagus planktovores.

a) Milkfish b) rainbow trout c) carp d) none of these

12. Energy is lost from the body of a fish in the feaces, urine, gill excretions and

as………………….

a) Heat b) sweat c) both d) none

13. Heat of nutrient metabolism, also called heat increment or…………………………,

which is the heat released by the many chemical reactions associated with the processing

of ingested feed.

a) specific dynamic action (SDA) b) heat release c) both d) none

14. …………….. is stored in the chemical structure of the complex molecules of feed

materials.

a). heat b) Energy c) both d) none

15. Fats are the ……………… of energy storage in plants and in animals.

a) principal form b) minimal form c) moderate form d) all these

16. In nature, carnivorous fish consume diets which are about ………………….. protein.

a) 30 percent b) 10 percent c) 2 Percent d) 50 percent

17. Digestion efficiency in fish ……………….. as feeding level is increased.

a) Equals b) decreases c) increases d) boosts

18. Starved fish are ………………than well-fed fish but there is always some expenditure of

energy for activity.

a) More active b) moderately active C) less active d) highly active

19. The low metabolic rate at low temperatures enables fish to survive for long periods

under ice where ………………..is available.

a) Greater food b) little food c) less food d) no food

 20. Simple proteins on hydrolysis yield only the amino acids and

occasionally…………………………………

a) L- carbohydrate b) small carbohydrate compounds c) small lipid compounds d)

large peptides

21. Example of conjugated proteins is:

a) Hemoglobin b) globulins c) albumin d) histones

22. The animal is incapable of synthesizing indispensable amino acids and must therefore

obtain these from the diet. These are called ………………….

a) Non-essential amino acids b) essential amino acids b) both d) none of these

23. Lysine is a ……………. amino acid

a) Acidic b) amphoteric c) basic d) all

24. Fish appear to utilize …………………. at various degrees of efficiency.

a) free amino acids b) derived amino acids c) both d) none

25. A typical example is the many weeks of migration by salmon in their return upstream to

spawn; ………………………….are burned for fuel to enable body processes to continue

during the strenuous journey.

a) Carbohydrates b) proteins c) stored lipid deposits d) all of these

True/false

1. Waxes are carbohydrates esters of long-chain alcohols. True /false

2. Linolenic acid would be written as 18:3w 3. True /false

3. Fish in general contain more w 3 than w 6 polyunsaturated fatty acids (PUFA) True

/false

4. The iodine value or degree of unsaturation of the oil was minimal in June and maximal in

April. True /false

5. Dietary linolenic acid or ethyl linolenate (18:3 w 3) gives a positive growth response for

rainbow trout. True /false

6. The positive effects of the oxidized fish oils were reversed by the addition of alpha-

tocopherol acetate or ethoxyoquin to the diet. True /false

7. universally digestible polysaccharides - principally starch. True /false

8. Glycogen is often referred to as plant starch because it is present in plants. True /false

9. free sugars such as glucose and fructose are termed monosaccharides. True /false

10. Carbohydrates of short chain lengths such as raffinose, stachyose and verbascose, are

classified as oligosaccharides. True /false

11. galactose and mannose occur only in free form. True /false

12. A general phenomenon, known as matstasis, is observed in a variety of pentoses and

hexoses as well as in certain disaccharides. True /false

13. Sugars containing the aldo or the keto group are capable of reducing copper in alkaline

solutions (Fehling's solution) to produce the brick-red colouration of cuprous ions. These

sugars are called reducing sugars. True /false

14. Glacatose is widely distributed in small amounts in fruits, plant juices and honey.

True /false

15. Galactose occurs in milk in combination with glucose. True /false

16. Dextrins are intermediate compounds resulting from incomplete hydrolysis or digestion

of glycogen. True /false

17. In carnivores such as the rainbow trout and sea perch, amylase is primarily of pancreatic

origin. True /false

18. The process of gelatinization involves both heat and alcohol. True /false

19. climatic factors also play an important role in carbohydrate metabolism in fish. True /false

20. The principal end-product of carbohydrate digestion, glucose, is metabolized in a manner

prevailing in all cells, i.e., via the reversible Emden-Meyerhoff pathway. True /false