INTRODUCTION

DEFINATION
• NUTRITION
A series of process: eating, digesting,
absorbing, tissue repair, and growth
 Food
• Something that can be eaten
 Feed
• Food for reared animals
• Process of giving food to animal
 Ration
• Food for a certain period of time, e.g, 24
hours.
 Diet

• Food prepared for a special objective

 Efficiency of fish VS land animals
in term of ENERGY use
• Release ammonia
• Fish density same as water
• Some fish has wind bladder for buoyancy
• Poikilothermic
 Anatomy of Digestive Tract
• Dietary stand point:
• Fish without stomach
• - carp, tilapia etc
• Fish with stomach
• - salmonids, catfish, eel etc
• Stomachlees fish:
• - no teeth in oral cavity
• Stomach
• - teeth in varying degrees
• - assist in the process of food capture
• - food swallowed as whole
• Pharyngeal teeth in carp
• - to compress food slightly
• Piranha
• - teeth help in capturing food

• Fish do not have salivary gland in oral

cavity
• Oesophagus – short, readily expanded
• Intestine:
• - catfish - one third of body length
• - tilapia - up to three times of its body
length
• Intestine of cyprinid:
• -poorly differentiated:
• 1. Anterior section
• 2. Proximal portion of midgut/small
intestine
• 3. Distal portion of midgut or small
intestine
• 4. Terminal section (hindgut) or rectum
• Pyloric caecae:
• - found in some fish – enlargement of
stomach surface
• Rectum
• - no clear defined rectum in fish.
• - closure of intestine is effected by the
anal sphincter
 General Overview on Dietary Requirements of
Fish Nutrition

6 major classes of nutrients :

Proteins
1) Proteins Lipids (certain types) Used as structural
Minerals material
2) Lipids Water

3) Carbohydrates
Carbohydrates
Oxidized to provide
Lipids
4) Vitamins energy
Proteins

5) Minerals
Trace minerals Function as essential
Vitamins components of coenzymes
6) Water in bio-chemical systems
 Protein
• Important component of animal tissue
• All cells synthesize protein in their life
cycle
• Protein must be included in diet to allow
normal growth.
• Some animal can synthesis protein from
non-protein N by microflora in intestine
• % protein requirement in diet of young
animal are highest, but declines gradually to
maturity when enough protein to maintain
body tissues is required.
• Productive function, will increase protein
requirement - pregnancy, lactation, egg
production etc
 What are proteins?
Proteins are polymer of amino acids (a.a)

a.a1 a.a2 a.a3

a.a4 a.a5
a.a6
a.an a.a7

Contains about 50%C, 22%O2, 7%H & 16%N

Proteins are essential ingredients in diets as source of

amino acids.
 What are Amino Acids?
• As their name, “Amino Acids” are composed of the
amino-group (-NH2) & carboxyl-group (-COOH)
 Can display reactions which are either acidic (from
COOH) or basic (from NH2)

O O
H2N C C OH
Carboxyl group
Amino group
R (acid)
(base) Side chain
Amino acids (continued)
Based on chemical structure of side group (R), a.a are classified into
following groups:
1) Aliphatic a.a (eg. glycine) 4) Heterocyclic a.a (eg. tryptophan)
O O
O O
H2N C C OH C
H2N C OH
H CH2
N
2) Aromatic a.a (eg. phenylalanine) H
O O
5) Acidic a.a (eg. aspartic acid)
H2N C C OH
O O
CH2 H2N C C OH
CH2 COOH
3) Sulphuric a.a (eg. cysteine)
6) Basic a.a (eg. lysine)
O O
O O
H2N C C OH C C
H2N OH
CH2 SH H2N CH2 CH2 CH2 CH2
Amino acids (continued)

• In general, there are 20 a.a that are found in proteins

(some proteins are devoid of some a.a; other contain
all 20 a.a).
• Physiologically, amino acids can be classified as:

1) Essential :
 Those a.a that body is not able to make
 Needed in diet
2) Non-essentials :
 Those a.a that body is able to make
(synthesize in the body)
Amino acids (continued)

Essential and Non-Essential Amino Acids

Essential (EAA) Non-Essential (NEAA)

Arginine (Arg) Alanine (Ala)
Histidine (His) Aspartic acid (Asp)
Isoleucine (Ile) Glutamic acid (Glu)
Leucine (Leu) Glycine (Gly)
Lysine (Lys) Hydroxyproline (Hyp)
Methionine (Met) Ornithine (Orn)
Phenylalanine (Phe) Proline (Pro)
Threonine (Thr) Serine (Ser)
Tryptophan (Trp) Tyrosine (Tyr)
Valine (Val) Cystine (Cys)
• Naturally occuring a.a are in L-
configuration.
• Synthetic a.a are D-form
• L-form are biologically more active
Amino acids (continued)
However, we should understand that:

Although NEAA are not dietary essential nutrients, they perform many
essential functions at cellular & metabolic level. In fact, it is often quoted
that NEAA are physiologically so essential that body ensure an adequate
supply by synthesis

From feed formulation viewpoint, it is important to realize that NEAA like

cysteine & tyrosine are synthesized within the body from EAA methionine
& phenylalanine, respectively

Met   Cys / Phe   Tyr

Thus:

if concentration of EAA in diet   synthesis of NEAA

 Protein Elongation

In protein molecules, a.a are joined together by peptide bond

a.a1 a.a2 a.a3

a.a4 a.a5

Peptide bond a.a6

a.an a.a7

**Peptide bond is a very strong bond

 Since a.a are soluble in water, with the elimination of one
molecule H2O,unite to form a peptide molecule leading
to the formation of typical peptide compounds
O O O O
H N C C OH + H N C C OH

Amino acid1
H R1 H R1 Amino acid2

H2O

O O O O
H N C C N C C OH dipeptide

H R1 H R1
 Combination of amino acids call as “peptide”
 2 a.a = dipeptide
 3 a.a = tripeptide
 4 – 10 a.a = oligopeptide
 > 10 a.a = polypeptide

Large polypeptide = PROTEIN MOLECULES

 Besides the PROTEIN MOLECULES, there are also PROTEIDES
which contain, apart from amino acids, other foreign
components in the loosely bound

a.a1 a.a3 a.a1 a.a2 a.a3

a.a2 a.a5
a.a4 a.a5 a.a4

a.a6 Foreign components a.a6

a.an a.a7 (eg. Carbohydrates, x x x

lipids, nucleic acid or
dyes)
PROTEIN MOLECULES PROTEIDES
• phosphoproteins Eg.
• albumin
• Lipoproteins (+lipids)
• globulin soluble protein
• myosin • nucleoproteins (+nucleic acids)
• fibrinogen • chromo-proteins (+dyes)
• keratin insoluble • certain enzymes
• collagen fibrous protein
Microscopy of keratin filaments inside cells. (In hair, nail
and skin cells of human)
What are the physiological functions of proteins?

Fish Body (Wwet) : 75% H2O, 16% Protein, 6% Lipid & 3% Ash

Protein are make up about 65-75% of the dietary dry weight

of many animals including fish and shrimp
Some examples of the importance of protein at
cellular level:
1) Protein are essential component of cell nucleus (eg.
Chromosomes are made up of protein & DNA)

2) All enzyme are protein

eg. Glucose        CO2 + H2O + Energy
      
Enzymes
Catalyze the biochemical reaction

3) Biological membranes (such as plasma membranes) are

made up to 30-60% proteins.
Some examples of the importance of protein at
macroscopic level:
1/3 protein

1) Formation/maintenance of muscle, skin, bones

2) Hormones like FSH, LH, GH and insulin

3) Antibodies (immune system)

4) Carrier proteins such myoglobin in hemoglobin (for O2 transport)

General structure of hormone (eg. Tryptophan)

Function of insulin (C257H383N65O77S6)

3D structure of myoglobin
Do fish & shrimp need dietary protein ?
All animals, including fish & shrimp need dietary
proteins for following reasons:
1) To get essential amino acids (those a.a. not made by body)

2) To use protein (a.a from protein) as energy  especially for

aquatic animals.

3) To use carbon skeleton of a.a to C6H1206

make other molecules like glucose
O O
4) To utilize amino group N in non- H2N C C OH
protein compounds such as purine
& pyrimidines (RNA, DNA) H

DNA, RNA
Nucleotides… the structural units of RNA, DNA,
How much dietary proteins fish need?

This is not easy to answer because protein requirement is:

1) Different for different sp.

2) Even for one sp., it depends on various biotic (age &

size) and abiotic (temp, salinity) factors.
 Some examples of dietary protein needed :
Size & Age Temperature Salinity
Channel catfish Chinook salmon Fingerling rainbow trout
Fry  40% 8C  40% 10ppt  40%
Fingerling  30-35% 15°C  55% 20ppt  45%
Large (>10g)  25-35%
Striped bass
Tilapia 20C  47%
<1g  40% 24°C  55%
Younger animals
1-5g  30-35% generally require higher
5-25g  25-35% In general protein levels of protein to
>25g  20-25% requirements for fish support their growth than
ranges from 25 – 60% of older animals:
Wdry
Shrimp Feed formulation for
<0.5g  45% Omnivores : 25 – 35 % larvae, fry & juvenile
1-5g  40% stages contain 5-10%
Carnivores : 40 – 60%
more protein than grower
5-25g  30%
diet formulated for
>25g  36% older/larger fish.
Protein needed (cntd)

Protein requirement of different cultured sp.

 Estimation of Protein Requirement

Since the protein sources are generally costly, in

commercial fish farming the primary concern is to find
the minimum dietary protein content (optimum level)
with the high rate of growth and satisfactory feed
utilization.
 Estimation of Protein Requirement (cntd)

The most common method : measuring the growth of fish fed diets containing
graded levels of protein  produced a “protein dose-response” curve
Accurate method: Quadratic regression

8.0 Quadratic regr.

Low er 95%
7.5 Upper 95%
Horizontal to max
7.0
Horizontal to Low er 95%

6.5

6.0
Fina l we ight (g)

5.5

5.0

4.5 Xeconomic
4.0

3.5
Xmin
3.0 Xmax
2.5

2.0
15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55

Die ta ry prote in (%)

However, note that the dietary proteins must contain a well
balanced mixture of both EAA & NEAA for optimum growth.

General dietary EAA requirement for most culture fish:

Essential (EAA) Requirement (g/100g Protein)

Arginine (Arg) ------------------------ 3.3 – 5.9
Histidine (His) ------------------------ 1.3 – 2.1
Isoleucine (Ile) ------------------------ 2.0 – 4.0
Leucine (Leu) ------------------------- 2.8 – 5.3
Lysine (Lys) -------------------------- 4.1 – 6.1
Methionine (Met) ------------------- 2.2 – 6.5
Phenylalanine (Phe) --------------- 5.0 – 6.5
Threonine (Thr) --------------------- 2.0 – 4.0
Tryptophan (Trp) -------------------- 0.3 – 1.4
Valine (Val) --------------------------- 2.3 – 4.0
 In some cases, although the crude protein content in the diet is high,
deficiency of some EAA was resulted extremely bad effect, such:

1) Stunted growth

Glutamic acid (NEAA) -deficient

Arginine (EAA)-deficient

Leucine (EAA)-deficient
NEAA

EAA
2) Deformity

Such scoliosis  deficiency syndrome for tryptophan in salmon

 What does happen to dietary proteins after it is
consumed by fish?

In short, proteins are digested to amino acids, and then amino

acids are absorbed & metabolized.

Protein digestion : breakage of peptide bonds by proteolytic

enzymes (also called protease) which takes place in
stomach & intestine of fish.

a.a1 a.a2 a.a3 a.a1 a.a2

a.a4 a.a5
a.a5
a.a4
Dietary proteins a.a6
a.a3

a.a
       a.an a.a7
a.a6
(polymer) n a.a7
Proteolytic enzymes
(proteinases, peptidases) Free amino acids
from stomach and pancreas (monomer)
 Digestion, absorption and transportation of protein nutritional
Secrete by stomach
STOMACH INTESTINE/CAECA
Pepsin Trypsin,
chemotrypsin,
     elastase, Secrete by pancreas
Protein caboxypeptidase,
a.a1 a.a2 a.a3
a.a5
aminopeptidase
a.a4
ORGANS
a.a6     
a.an a.a7 Peptide  brain
a.a5 a.a6
 heart
a.a7
 muscles
Peptide a.a1 a.a2 a.a3  etc
a.a4
a.a5 a.a6 a.a7

a.a1 a.a2 a.a3 Amino Acids

a.a4
a.a5 a.a6 a.a7
a.a4
a.a1 a.a2 a.a3
BLOOD
VESSEL
Amino acids metabolism:

Amino that taken from blood are used as :

1) Energy
Energy
O O ATP

H 2N C C OH      CO2 + H2O
R NH3 either used as source
of N or excreted

This process call as protein degradation (the simples

protein (amino acids) are break to smaller molecules).
2) Incorporated into new proteins or a.a derivatives like T3 & T4
a.a1 a.a2 a.a3 Such as:
a.a5 a.a6 a.a7 a.a4 a.a5
 enzymes
a.a1 a.a2 a.a3
a.a4  a.a6  structural component muscle
a.an a.an a.a7
 hormones such GH
 etc

This process call as protein synthesis (or protein turn over)

Animal is able to grow when the rate of protein synthesis is

greater than the rate of protein degradation.

Conversely, if there is insufficient energy, then protein

degradation will be greater than protein synthesis, animal will
lose weight.

Tyrosine Triiodothyronine Thyroxine (T4)

(T3)
 Evaluation of Protein Sources

From nutritional point of view, selection of material as protein source in

feed formulation must be concerned on:
1) Digestibility of the protein
2) Nutritional value of protein (protein utilization)
Evaluation of Protein Sources (cntd)

Measurement of Protein digestibility

A distinction must be made between the true and apparent digestibility

of protein:

Apparent digestibility = feed N – faecal N X 100

feed N

However, N in the faecal may also come from the metabolic activity
(known as endogenous faecal N – EFN) 
resulted too low digestibility if estimated by this way

EFN can be quantified with the aid of a protein-free diet

Evaluation of Protein Sources (cntd)

Taking the fraction of EFN into account, the true protein

digestibility is obtained from the equation:

True digestibility = feed N – (faecal N – EFN) X 100

feed N
 Factors that effected on the digestibility in fish:

1) Species
(digestibility of herbivores/omnivores fish > carnivores fish –
especially on plant proteins)

2) Protein Source
(digestibility on animal proteins > plant protein – especially in
carnivores fish)

3) Age / size
(digestibility of younger fish < adult)

4) Temperature
(digestibility in higher temp. > lower temp.)
Digestibility in different species



Digestibility in different protein sources

Animal
Protein

Plant
Protein
Digestibility in different age/size
Digestibility in different temperature
Evaluation of Protein Sources (cntd)

Measurement of nutritional value of protein

Since proteins are also used as source of energy, dietary protein is

put to optimal use when as much as possible of the protein ingested
is used for synthesis of tissue proteins (for growth).

Nutritional values of proteins are used as a guide to the effectiveness

of particular protein source in supplying the animal’s requirement for
growth.
Evaluation of Protein Sources (cntd)

Measurement of nutritional value of protein (cntd)

There are 3 main methods for quantifying the nutritional value of

proteins :

1) Protein Efficiency Ratio (PER)

PER = g wet weight gain
g crude protein fed

PER   good protein utilization  protein value 

However, this method assume that the protein concentration of the

fish as a whole does not change during the course of experiment
(not necessarily be true in all cases) and need long test period (not
less than 2 – 3 month).

More meaningful results may obtained by measure the Net Protein

Utilization (NPU)
Evaluation of Protein Sources (cntd)

2) Net Protein Utilization (NPU)

NPU = biological value (BV) X digestibility

where BV = N gain + Endogenous N + EFN X 100

Absorbed N
NPU = Final body protein (g) – initial body protein (g) X 100
Total protein fed (g)

NPU   good protein utilization  protein value 

Evaluation of Protein Sources (cntd)

3) Essential Amino Acid Index (EAA-index)

Can only be used if the amino acid requirement of the tested species is
known.

The nutritional value of a protein can then be made by directly

comparing the EAA content (in protein sources) with the EAA required
by the species.

Eg: When estimates EAA index of whole chicken eggs

EPR (egg protein ratio) = . % amino acid needed X 100

% amino acid in egg protein

the EAA-index of the EPRs for 10 EAA is:

EAA-index = 10
(EPR1 X EPR2 X EPR3 X ……EPR10)