Aquaculture of Indigenous Mekong Fish SP

Component Report No.
20
Aquaculture of Indigenous Mekong Fish Species
Training in Feeds and Feeding for Indigenous Mekong Fish Species

February 24-27, 2004, Udon Thani, Thailand
CONSULTANTS REPORT
Amonrat Sermwatanakul, Nantiya Unprasert Mali Boonyaratpalin and

Pisamai Somseub
Bangkok, May 2004

The justification for the Component rests on the threat to biodiversity posed by introduction of exotic
fish species for aquaculture and culture-based fisheries. Thus, the aim of the Component is to develop
economically feasible aquaculture techniques for 5-8 indigenous Mekong fish species, to complement
or replace the use of exotic species. This will be achieved by on-station as well as on-farm research
on breeding, nursery and grow-out techniques for the selected species, followed by elaboration of
extension materials and packages. The immediate target group for the component is Government
researchers under the aquaculture research and development institutions in the riparian countries,
while the development target group is small-scale farmers.
Contact address:
AIMS, P.O. Box 7035, Vientiane, Lao PDR
Telephone: +856 (21) 223436
Fax: +856 (21) 223610
E-mail 1: fip@laopdr.com
E-mail 2: niklas@laopdr.com
2
TABLE OF CONTENTS
Background ....................................................................................................................4
Outline of the Course and Schedule...............................................................................4
Results of the Workshop................................................................................................5
Recommendations..........................................................................................................6
Conclusion .....................................................................................................................6
Annex 1. Terms of reference .........................................................................................7
Annex 2. Background on fish nutrition of participants................................................12
Annex 3. Digestive systems in fish..............................................................................15
Annex 4. Research plan on-farm made feed................................................................17
Annex 5. Research plan on feed and feeding in Cirrhinus microlepis ........................18
Annex 6. List of Participants .......................................................................................23
Annex 7. Lecture Notes ...............................................................................................27
3
Background
The progress has been made in the breeding of indigenous Mekong fish and seed
is now being produced for several species. However, the nutritional requirement of
these species remains to be poorly understood. Also there are problems in terms of
egg and sperm quality leading to poor early development. An ensuing compromised
reproductive success is thought to be at least in part related to a poor nutrition. The
present short course was therefore conducted primarily with the following objectives:
1) to provide basic knowledge about nutrients and their interaction, 2) to introduce to
the basic composition, utility, and constraints of some of the locally available major
feed ingredients, 3) to demonstrate the feed production technique, and 4) to elaborate
about the feeding practice pertaining to various life stages of some of the important
indigenous fish species of the Mekong basin. The additional foci of this course were
the subjects of fish physiology, fish health, and food biochemistry as co-essentials for
a successful aquaculture.
Outline of the Course and Schedule

The course was organized around the key points pertaining to the process of
digestion in the gastrointestinal tract, basic nutrient requirements, and how to make
fish feed locally on a small scale.
Nutrient requirements were described with respect to protein, fat, carbohydrate,

vitamins, minerals and energy. These requirements vary with fish specie as digestive
system anatomy and physiology are different among species depending mainly on the
feed type. In general, herbivores such as tilapia and carp have relatively small
stomachs and long intestines whereas omnivores have a moderate size stomach and
intestine. On the other hand, carnivore such as trout and striped bass possess a
relatively large stomach and short intestine.
Feeding regimes are also different among herbivorous omnivorous and
carnivorous species as can be described in terms of variations found in stomach
contents, natural food, supplementary feeds.
Feed formulation part of the program included introduction to nutrient

composition of common feed ingredients, feed additives, nutrient requirements, feed
composition, types of feed and how to compute to meet dietary needs. It also
included cost of feed, nutrient availability and minimum-maximum restrictions of
ingredient inclusion in the feed.
Proteins are composed of amino acids which may be described as essential

and non-essential. Essential amino acids must be provided in the feed. The following
amino acids have been shown to be essential for fish, arginine, histidine, isoleucine,
leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine. The
amount of each amino acid required varies from species and excessive quantities can
be detrimental to fish health. Shortage of amino acids can result typically in scoliosis
(curvature of the spine) in fish. Proteins are very important for the growth of fish and
research has shown that the growth is suppressed in protein deficiency. In the wild,
the free choice of food ensures enough protein. However, commercial feeds may lack
4
abundant protein since it is an expensive nutrient that may result in protein-energy
imbalance with impaired performance.
Carbohydrates are found in greater abundance primarily in plants and

carnivorous fish have problems digesting this class of nutrients. Lipids, on the other
hand, are frequently found in both plant and animal tissues and are readily digestible
in all fish species. Many herbivorous fish have symbiotic bacteria in their guts which
digest the carbohydrates thus liberating usable energy to the fish. Lipids provide much
more energy than do carbohydrates and they also supply fatty acids some of which are
used for the synthesis of biologically important compounds. Fats also provide for
major source of energy reserves in fish. . Predaceous fish normally have
comparatively very high growth rates since their diet is composed of live fish, which
are naturally high in lipids. The importance of protein and lipids as energy sources,
becomes quite apparent during periods of starvation. The quantities of both proteins
and lipids are significantly reduced in starved fish.
Vitamins and minerals constitute a small proportion of fish feed and are therefore
regarded as micro-ingredients. However they play a very important role in the
utilization of feed, as components of the animal body, and in biological function.
Their deficiency results in poor performance and in many cases as characteristic
deficiency symptoms.
Vitamins can be classified as water soluble and fat soluble. Water soluble vitamins
include B group vitamins such as Thiamine (B1). Riboflavin (B2), Pyridoxine (B6),
Cyanocobalamin (B12), Niacin (B3), Pantothenic Acid (B5), Biotin, Choline, Inositol,
Folic Acid, and vitamin C. Fat soluble vitamins are vitamins A, D, E, and K.
Minerals can be grouped based on their dietary concentration and macro-minerals and
micro-minerals. Macro-minerals include Calcium, Phosphorus, Sodium, Potassium
and Magnesium. Micro-minerals include Iron, Copper, Manganese, Zinc, Selenium
etc.
Results of the Workshop

It is necessary for participants to review their basic knowledge in fish nutrition before
enrolling for the training course. The presence of a coordinator from each country
appears to be useful in terms of helping their group to understand .the instructions and
demonstrations. It was not clear however, the extent of availability of opportunity to
practice feed production by the trainees when they go back to their home. Overall, the
enthusiasm and interest of participants in the program appeared to be satisfactory.
The availability of major feed ingredients is also limited in the rural areas that
constitute the Mekong basin. Corn, rice bran, soybean meal, and broken rice are the
only few ingredients generally available. Vitamins and minerals are even more
difficult to obtain. Therefore, to formulate a nutritionally adequate feed remains a
challenging task in such circumstances.
It is not easy to comprehend the basics of fish nutrition and feeding management
within one week duration of the course. Therefore, the participants must learn more
from the books and other available literature.
5
Recommendations
It is understood that the participants hold responsibilities as government officers in the
fisheries departments. It is therefore suggested that participants wanting to formulate
diets in their home country should be encouraged to send the formula to one of our
staff for a double check and further advice. It is suggested in addition that in case of a
difficulty finding micro-ingredients such as vitamins and minerals in their locality,
they can order via the MRC Thailand. We can therefore help them formulate as well
as can place an order on their behalf.
Most of the participants had extension background and some experience in fish
breeding. It is therefore complementary for them to understand basics of nutrition and
how to formulate diets for broodstock and larvae. It would therefore be of value if
formulation and preparation of diets could be included under the technology transfer
program. A simple feed production unit comprising a grinder, mixer and pelleting
machine may cost some 200,000 baht. A spreadsheet could be developed for
computations. Those seeking advanced understanding in this respect should be helped
as a part of this program and may be provided additional training
The continuity of practice in the art of nutrition and feed preparation cannot be
overemphasized. To this objective and also to further broaden their knowledge base,
we offer that participants at their convenience visit our laboratory and practice making
the micro-ingredient premix, follow the feed production procedures, and as well gain
information on the analytical aspects pertaining to evaluating composition of the feed.
Conclusion
The Regional Workshop on “Feed and Feeding for Indigenous Mekong Fish Species”
was held during February 24-27, 2004 at Inland Fisheries Research Center,
Udonthani, Thailand. Twentyone participants from Cambodia, Lao PDR, Thailand
and Vietnam attended the workshop. The course included twenty one lecture hours
with six and a half hours allocated for practice and farm work. The program’s main
goals were to provide for an introduction to the nutritional needs of the fish and the art
and science of practical feed production and feeding management. An opportunity to
formulate practical feed was also provided to the participants.
6
Annex 1. Terms of reference
“Training in feeds and feeding for Indigenous Mekong Fish Species”
Background
The status of feeds and feeding for inland aquaculture in the Mekong basin was recently
reviewed by Peter Edwards and Geoff Allan 1 . The report identifies researchable issues,
summarized as follows:
1. Optimal combinations of fertilizers feed ingredients and manufactured feeds to
maximize production of different species and life stages for different regions.
2. Development of time and cost effective feeds for grass carp
3. Development of cost-effective diets for high-value species like giant gourami,
climbing perch and Spinibarbus denticulatus.
4. Determination of optimal methods of preparing .e.g., grinding, cooking,
binding, and delivering feeds.
5. Understanding the supply and demand characteristics of trash fish, both
freshwater and marine, in order to reduce dependence on this resource where
necessary.
6. Understanding the variability in supply and composition of rice bran and
evaluation of the potential to increase use of this ingredient in feeds.
7. Improving farmer extension on feeds and feeding
8. Improving nutritional research capacity through networking and training.
AIMS has made progress in the breeding of indigenous Mekong fish species, and seed
is now being produced for several species. However, some of the species are new in
terms of aquaculture, and their nutritional requirements are poorly understood.
Problems have been encountered in conditioning of broodstock for spawning, with poor
egg and sperm development, and the feed composition is a prime candidate factor.
Nursing and grow-out faces analogous problems.
Commercial feeds are available, but generally have not been developed for the
indigenous species now being developed for aquaculture. Generic commercial feeds
may also be too costly.
Objectives
The training will focus on sharing of knowledge on feed and feeding of indigenous
Mekong fish species at different life stages. After the training the participants should
• Simple approaches to assessing nutritional requirements

be familiar with:
• The basics of natural food production in ponds

• Techniques for on-farm production of feeds
• Different techniques for delivery of feeds
Outputs
The outputs will consist of:
1
Edwards P. and Allan G. (2002). Review of feeds and feeding for inland aquaculture in Mekong
region countries. ACIAR draft report presented at a workshop in Siem Riep.
7
• A training/workshop on feeds and feeding planned and approved by the
• The above training carried out

Component Coordinator
• A report to the Coordinator,

(a) an outline of the course plan and schedule,
(b) recommendations on the implementation of research plan for the
Component, with particular attention to profitability for small scale farmers,
(c) recommendations for follow-up training, including draft ToR and
(d) an evaluation of the course by the participants, including their ideas for
• A presentation (e.g. PowerPoint) covering the main outcomes of the training

future training
The draft report will be handed over to the AIMS Coordinator at a final debriefing at
the end of the training. A final report and the presentation will be forwarded to the
AIMS Coordinator latest three weeks after the end of the training.
SCOPE OF WORK
Training
• Participants presented their ongoing or planned research on feed and feeding

The scope of the training comprised to the following:
• Presentations by trainer(s)
• Practical work, including making on-farm feed
• Discussion and elaboration of research topics (working groups, focussing on
the participants own research) and approaches for extension to farmers,
including on-farm research.
• Prepare the final report as described under 'Outputs'

Reporting
Implementation of the training event

An expert in feeds and feeding of fish, with considerable experience in training and
working with small-scale farms and local hatcheries will plan and carry out the
training. Experience with aquaculture and inland fisheries in Southeast Asia is
essential.
The consultancy was carried out under the overall supervision of the Fisheries
Programme Management and the Country Offices, under the direct supervision of the
Component Coordinator and National Component Directors for the Component, and
in close cooperation with other national and international staff of the Fisheries
Programme.
The Component provided logistical support as well as secretarial support, office

space and other necessary assistance as needed.
8
The total duration for the consultancy is outlined below (tentative dates).
Activity Days Start Finish

Preparation of the 3
training/workshop
Carry out Training/workshop 4 24 February 27 February 2004
2004
Report preparation 27 February 2004
Total 7
DSA and assignment-related expenses, such as materials for the workshop and duty
travel will be reimbursed against travel expense claim(s) and receipts.
Course Description : Introduction to fish nutrition, fish feeds, feeding fish, feeding
systems and feed management and fish feeds
Course outline
1 Introduction to fish nutrition 3.5

2 Fish feeds 3
3 Feeding fish 3.5
4 Feeding systems and feed management 3
5 Feed formulation practice 6.5
6 Formulating your own feed 3.5
7 Discussion 4.5
27.5
Lectures and discussion= 21 hrs., Practicum 6.5 hrs.
Number of participants: 15-20
Lecturers:
Name Qualification Affiliation

Mali Boonyaratpalin Ph.D. Dept. of Fisheries
Amonrat Sermwatanakul Ph.D. Dept. of Fisheries
Nanthiya Unprasert Ph.D. Dept. of Fisheries
Pisamai Somsueb M.Sc. Dept. of Fisheries
9
Tentative Schedule
Tuesday, February 24, 2004
08:30-12:00 Introduction to Fish Nutrition

- Nutrients (protein and amino acids, lipid, carbohydrate, vitamins and minerals)
and energy
- Nutrient requirements (growth, reproduction and health)
- Species differences
- Other dietary components (pigments, feeding stimulants etc.)
- Nutrition and product quality
- Nutritional diseases
(Dr. Amonrat Sermwatanakul)
12.00-13.00 Lunch
13.00-16.00 Fish Feeds

- Feed formulation
- Feed types
- Feed manufacturing technology
- Special feeds
- Feed quality (Composition and chemical analyses, rancidity etc.)
- Feed storage
(Dr. Nanthiya Unprasert)
16.00-16.30 Discussion Period
Wednesday, February 25, 2004
08:30-12:00 Feeding Fish

- General concepts
- Environmental factors and food consumption
- Feeding behaviour
- Techniques for measuring food intake
- How much to feed?
- Feeding larval fish
- Broodstock Feeding
(Pisamai Somsueb)
12.00-13.00 Lunch
14:00-16:00 Feeding Systems and Feed Management

- Feed planning and production plans
- Distribution of feed
- Feeding techniques (hand feeding, demand feeding and other techniques)
- Feeding systems
10
Thursday, February 26, 2004
08:30-12:00 Fish Feed Formulation Practice

- Ingredient types
- Optimum ratio of each ingredient
- Premix calculation
- Set up your own worksheet
- Practicing your own worksheet
Please send all ingredients to be analyzed at DOF feed quality control laboratory 2
weeks ahead.
12.00-13.00 Lunch
13:00-16:30 Open Discussion

(Dr. Mali Boonyaratpalin and staff)
Friday, February 27, 2004
08:30-12:00 Formulating your own feed (lecture)

12.00-13.00 Lunch
13:00-16:00 Fish Feed Formulation Practice (continued from Feb 26)

Breaks: 10.00-10.15 and 14.30-14.45
11
Annex 2. Background on fish nutrition of participants
“Training in Feeds and Feeding for Indigenous Mekong Fish Species”
Home Work
February 24, 2004
____________________________________
1. Name…………………………………………………………………………………
2. Position………………………………………………………………………………
3. Job
description……………………………………………………………………………
4. Have you ever been involved in fish nutrition?

…………………………………………………………………………………………
…………………………………………………………………………………………
………………
If yes. Please describe.
…………………………………………………………………………………..………
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5. What are nutrition problem do you have?
…………………………………………………………………………………………
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6. Have you ever been conducting fish nutrition or feeding research?
…………………………………………………………………………………………
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If yes. Please describe..
…………………………………………………………………………………………
…………………………………………………………………………………………..
…………………………………………………………………………………………
………………………
7. In case, you are in charge of extension official.
7.1 What kind of fish you are responsible?
…………………………………………………………………………………………
…………………………………………………………………………………………..
7.2 What do you want to know about feed and feeding?
…………………………………………………………………………………………
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8. What ingredients you have used? What are those formulas?
…………………………………………………………………………………..………
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9. What is the actual feed or ingredients that the farmer use to grow the fish?
…………………………………………………………………………………………
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10. What are their problems involving with feed used?
…………………………………………………………………………………………
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13
Summary Background Information from Participants
Cambodia Loa PDR Thailand Vietnam

No. of 4 4 6 5
Questionnaires
Position National Coordinator National Coordinator Fisheries Biologists Aquaculturist
Field Coordinator, Technician, Researcher,
Researcher, Technician
Technician,
Area of Job Researcher (2) Fish Breeder Research

Extension (2) Researchers
Nutrition No. (2) Yes No (3) Yes (5)
Background Yes (2) Yes (3)
Fish Species Pangasianodon hypophthalmus Tilapia Giant catfish Tilapia,
involved Barbonymus gonionotus Common carp Tilapia, carp, catfish
Trichogaster pectoralis Grass Carp B. gonionotus
Leptobarbus hoevenii Silver Barb C. microlepis
Ingredients used Fish meal, Rice bran, Broken Rice bran, Broken rice, Soybean Commercial feed, cassava, Soybean, Rice bran, Fish meal,
on farm rice, Soybean, Mung bean, meal, Corn, commercial feed, Beer rice bran, corn meal, Chicken Broken rice, Cassava, Duck weed,
Rice, Vegetable, Corn, Trash waste offal, Aquatic plant, termite Corn
fish, Lemna, Termite, Rice vine
waste
Problem in feed 1. Lacking knowledge on a 1. Non palatability feed 1. Poor performance 1. Fish nutrition requirement
and feeding growing Leptobarbus 2. Poor performance 2. Expensive feed
2. Poor feed performance 3. Immature broodstock 3. Poor performance
3. Immature broodstock 4. Insufficient knowledge of feed
4. Feed formulation knowledge formulation and requirement
5. Immature broodstock
14
Annex 3. Digestive systems in fish
Assignment for Group Work
February 26, 2004
____________________________________
Objectives:
To learn about the various digestive system in fish.
To identify the difference of fish digestive system.
Activities:
Each group of participants be able to identify the difference of digestive
system:
- Study on the variety of fish mouth position
- Measure the body and intestine length
-
- Identify the fish as herbivorous / Carnivorous /
Omnivorous
-
Each group prepared their own farm made feed as follows.
15
Result of four fish species digestive tracts difference
Colour of
Total internal
Position of Intestine length abdominal Identification
Item Species the Mouth Teeth Stomach (cm) (cm) IL/TL wall as
Cirrhinus
microlepis, protracted stomach-
1 Pawn protusible no teeth less 491.5 47.3 10.39 dark Herbivore
Labeo rohita,
Indian carp, protracted stomach-
2 Rohu protusible no teeth less 353 40 8.83 gray Herbivore
canines teeth,
upper and
Channa lower teeth, J-shaped
striata, superior pharyngeal and 2
3 Snakehead mouth teeth appendices 21.7 32.5 0.67 white Carnivore
Oreochromis
niloticus, straight
4 Nile tilapia terminal villiform teeth stomach 121.5 27 4.50 dark Omnivore
16
Annex 4. Research plan on-farm made feed
February 26, 2004
____________________________________
Objectives:
To learn about fish feed ingredient.
To practice on making farm made feed.
Activities:
Each group of participants be able to understand fish ingredients:
- animal protein sources: fish meal, squid meal, squid liver meal,
shrimp head meal
- plant protein sources: soybean meal, corn gluten meal, wheat
gluten meal,
- lipid sources: soybean oil, fish oil, lecithin
- Binder: alpha starch, gluten, wheat, fresh banana
- Carbohydrate sources: rice bran, broken rice, wheat, cassava
meal
- Vitamins and Minerals
Each group prepared their own farm made feed as follow.
Ingredients 1 2 3 4
Fish meal 30 30 30 30
Soybean Meal 20 20 20 20
Rice bran Meal 24 19 15 36
Broken Rice 20 - - -
Vitamins 2 2 2 2
Minerals 2 2 2 2
Soybean Oil 1 1 1 1
Fish Oil 1 1 1 1
Cassava - 10 14 -
Alpha starch - 15 - -
Wheat - - 15 -
Wheat Gluten - - - 8
Total 100 100 100 100
17
Annex 5. Research plan on feed and feeding in Cirrhinus microlepis
February 26-27, 2004
____________________________________
Objectives:
To make a research plan on feed and feeding of Cirrhinus microlepis
Activities:
Each group of participants propose the research plan on feed and feeding of
Cirrhinus microlepis.
Each plan was discussed and recommended by consultants.
Corrected protocol of each plan will be submitted to MRC.
Project Proposal 1
1. Study stomach content in wild fish (Lao/Thai)
1.1 Review relevant reports and information (including species synopsis from
Sukumasavin and Leelapatra, 2000)
Proposal 1.1:
- Review/Desk study carried out by whom?
Proposal 1.2:
- Time frame: February 2004 - January 2005
- Collect 30 wild fish sample from fisherman and preserve digestive tract in
formalin
- Identify food items in gut
- Report
Proposal 1.3:
- Preliminary study on commercial catfish feed, 30% protein for Cirrhinus
microlepis broodstock with 1% feeding rate.
- Inspect egg development from March to May.
18
Project Proposal 2
Country: Cambodia and Vietnam
Tittle: Study on feeds and feeding regime for Cirrhinus microlepis broodstock
reared in earthen ponds
Output: Improved Quantity and Quality of Mature Broodstock of Cirrhinus
microlepis
Reared in Captive Conditions
Justification:
1. The Component has achieved the initial results of artificial propagation and nursing
of Cirrhinus microlepis. However, wild breeders are a main source of broodfish to
mass-produce larvae.
2. Some F1 fish have matured in earthen ponds, but they produced small number of
eggs or milt in compared with the wild breeders.
3. Feeds and feeding regime for broodfish may be the key factors affect to broodfish
maturation in captive environment.
4. A thick fat layer in broodfish’ abdominal cavity is often observed. Appropriate
feeds and feeding regime need to be studied to increase number of mature broodfish
and reduce fat content in their cavity.
Design :
1. Feeds:
- Farm made feed: Diets are formulated based on 5 main ingredients (fish meal, rice
bran, broken rice, soybean meal and fish oil) plus. (Vitamin C + vitamin E) = 0.1%.
- Three types of diets containing 25%, 30% and 35% protein is formulated
- Additional feed: monofilament algae and green bean germ
2. Feeding regimes:
- Feeding rate:
+ Jan. – Apr. (Before - spawning season): 2%
+ May – August (During spawning season): 1%
3. Experimental design:
Treatment 1 2 3
No. fish 30 30 30
Farm made 25% protein 30% protein 35%
feed
Additional feed described described described
above above above
Feeding regime described described described
above above above
- Each treatment of 30 fish, each >1.5kg, stocked into 300m2 pond in triplicate
(9 ponds)
- Water exchange 30% twice monthly during spawning season
- Fish are checked for maturity each month during spawning season.
4. Feed preparation
- Soybean is grinded and cooked first
- Add broken rice and rice bran and continue cooking
19
- Vitamin C and E are mixed with fish meal and added into cooked feed and mix
them together properly
- The cooked feed is formed in small piece and provided to brood fish through
feeding trey.
.
20
Project Proposal 3
Country: Nakorn Panom, Thailand
Title: Protein Requirement for Juvenile of Cirrhinus microlepis
Objective
• Study on the protein requirement for juvenile of C. microlepis
• Study on survival rate, growth rate, FCR, cost in nursing juvenile of C.
microlepis
Design
• Divide fish in 3 treatment with 3 replication
Treatment 1 feed with 20 % protein
(herbivorous feed)
(catfish feed)
(shrimp feed)
• The experimental fish start from 50 g and finish experiment when fish grow up
to
100 g
• Experiment in fibreglass tank 1.5 ton with stocking density 50 ind/ton
• The fish were fed 2 time/day by satiation
• Collect and analysis data for conclusion to optimum feed formula. From
experiment we assume that the best protein level is 30 %. We formulated feed
by using local ingredients to extend farmer
fish meal 67.0 kg
rice bran 16.5 kg
boiled broken rice 16.5 kg
Vitamin C 100 g
21
Project Proposal 4
Country: Udonthani, Thailand
Title: Dietary Requirement of Cirrhinus microlepis Broodstock

Output: Mature Broodstock of Reared in the earthen pond
Justification: What kind of feed that this kind of fish prefers to eat?
Design:
Experimental fish = 60, 6 years old, Cirrhinus microlepis average weight
1.5 kg in Udon Thani Inland Fisheries Research and
Development Center
Experimental unit = 9 pens in a 3200 m2 earthen pond

(each unit is about 320m2)
Experimental design = CRD with 3 treatments and 3 replications

Treatment 1 = Control no feeding (fish live on natural food)
Treatment 2 = 40% crude protein marine shrimp feed
Treatment 3 = 40% crude protein farm made feed.
Fish meal
Rice bran
Wheat starch
Spirulina
Vitamin C
Vitamin E
Fish oil
Experimental management
- Feed one a day at 1.5% of body weight per day, 6 day a week
- Fertilize the whole pond by add chicken manure to keep the
transparency at 30 cm
- Environmental stimulation by 50% water exchange every two week in
May and June
Experimental data collection:
June check maturation of male and female every 2 week, by
- Make by stripping at observe sperm if they are under microscope
- Record number of male or female with egg or sperm and quality of
sperm and egg or stage
- Artificial breeding if egg and sperm are ready then percent
fertilization, hatching rate and survival are conducted.
22
Annex 6. List of Participants
Regional Workshop on :
February 24-27, 2004
Udonthani Inland Fisheries Research and Development Center, Thailand
Name Country
List of Participants
1. Mr. Ouk Vibol Cambodia
2. Mr. Ros Narin Cambodia
3. Mr. Lim Ngeth Cambodia
4. Mr. Hang Savin Cambodia
5. Mr. Somboun Lao PDR
6. Mr. Udeth Lao PDR
7. Ms. Somphouthone Lao PDR
8. Mr. Nantha Lao PDR
9. Mr. Sisouphan Lao PDR
10. Mr. Decha Rodrarung Udonthani, Thailand
11. Ms. Nongyao Manee Yasotorn, Thailand
12. Ms. Pongsai Chansri Surin, Thailand
13. Ms. Ratana Insiripong Khon Kaen, Thailand
14. Mr. Chainarong Chunchom Kalasin, Thailand
15. Ms. Wirawan Rayan Sakorn Nakorn, Thailand
17. Mr. Huynh Huu Ngai Vietnam
18. Mr. Thi Thanh Vinh Vietnam
19. Mr. Hoang Quang Bao Vietnam
20. Mr. Nguyen Trong Hien Vietnam
21. Mr. Nguyen Thanh Trung Vietnam
23
Name Country
List of Lecturers
1. Dr.Mali Boonyaratpalin Thailand
2. Dr. Amonrat Sermwatanakul Thailand
3. Dr. Nantiya Unprasert Thailand
4. Mrs. Pisamai Somsueb Thailand
MRC
1. Dr. Niklas Mattson MRC
24
Tentative Schedule
Tuesday, February 24, 2004
08:30-12:00 Introduction to Fish Nutrition

- Nutrients (protein and amino acids, lipid, carbohydrate, vitamins and minerals) and
energy
- Nutrient requirements (growth, reproduction and health)
- Species differences
- Other dietary components (pigments, feeding stimulants etc.)
- Nutrition and product quality
- Nutritional diseases
12.00-13.00 Lunch
13.00-16.00 Fish Feeds

- Feed formulation
- Feed types
- Feed manufacturing technology
- Special feeds
- Feed quality (Composition and chemical analyses, rancidity etc.)
- Feed storage
Wednesday, February 25, 2004
08:30-12:00 Feeding Fish

- General concepts
- Environmental factors and food consumption
- Feeding behaviour
- Techniques for measuring food intake
- How much to feed?
- Feeding larval fish
- Broodstock Feeding
(Pisamai Somsueb)
12.00-13.00 Lunch
14:00-16:00 Feeding Systems and Feed Management

- Feed planning and production plans
- Distribution of feed
- Feeding techniques (hand feeding, demand feeding and other techniques)
- Feeding systems
25
Thursday, February 26, 2004
08:30-12:00 Fish Feed Formulation Practice

- Ingredient types
- Optimum ratio of each ingredient
- Premix calculation
- Set up your own worksheet
- Practicing your own worksheet
Please send all ingredients to be analyzed at DOF feed quality control laboratory 2 weeks
ahead.
12.00-13.00 Lunch
13:00-16:30 Open Discussion

(Dr. Mali Boonyaratpalin and staff)
Friday, February 27, 2004
08:30-12:00 Formulating your own feed (lecture)

12.00-13.00 Lunch
13:00-16:00 Fish Feed Formulation Practice (continued from Feb 26)

Breaks: 10.00-10.15 and 14.30-14.45
26
Annex 7. Lecture Notes
FOR TRAINING PURPOSE ONLY
Lecture Notes
Training in Feed and Feeding

for Indigenous Mekong Fish Species
24-27 February 2004
Feed Formulations
Dr. Nanthiya Unprasert
Aquatic feeds Quality Control and Research Group

Inland feed Research Institute
Inland Fisheries Research and Development Bureau
Department of Fisheries, Bangkok 10900
Thailand
27
Fish Feed Formulations
Introduction
Formulation of well-balanced diets and their adequate feeding are the most important
for the successful of aquaculture practice. With intake of adequate feeds, fish is able to
maintained its health and be productive, regardless of the quality of its environment.
Formulation is the identification and evaluation of nutrients in a diet so that all necessary
dietary nutrients are supplied in optimum quantities to support the maintenance, growth and
production. Knowledge of nutritional requirements, feed ingredient sources, availability,
nutrient content, digestibility, cost, feeding behaviour and digestive capacity of cultured
species are needed to produce efficient diets. There is also a need to understand that
supplying adequate nutrition for various species involves not only the formulation of diets
containing desired essential nutrients but also the proper management of numerous factors
relating to diet quality and intake. The bioavailability of nutrients, diet palatability or
acceptability, feed manufacture, storage methods and chemical contamination may have
significant effect on fish performance.
Nutrient Requirement
Qualitatively, 40 nutrients have been identified as necessary for the normal
metabolic function on fish (Robinson, 1991). The quantitative requirements for most
nutrients have been identified for fish. Nutritional requirements for most fish have generally
been based on studies with small fish conducted under conditions presumed to be near
optimum; the requirement being based primarily on weight gain and feed efficiency. The
production of nutritionally well balanced diets for fish requires research, quality control, and
biological evaluation. Fig.1 outlines the various factors that affect successful feed formulation
and development of feeding program for culturing fish in captivity. (Cho et al., 1985)
Nutritional deficient diets impair fish productivity. The borderline between
reduced growth and diminished health is always very difficult to define. The result in
deterioration of health is recognized when diseases ensue. The ability to recognize a
reduction in growth and health performance during its initial stages and the corrective action
will remain an essential part of the skill of the fish culturist. Diets with nutrient deficiencies
can also negatively influence growth of fish. Balances of essential nutrients at an adequate
proportion, not only results in higher production but also provides the nutrients necessary to
promote recovery from disease or help the fish in overcoming the effects of environmental
stress. Hence, nutritionally balance and quality controlled diets are of critical importance for
aquaculture.
The dietary requirements can be established for energy, protein and amino
acids, lipids, minerals and vitamins. The requirement values represent minimum amounts for
maximum performance of fish under experimental conditions. The requirement values do not
28
include any surpluses. In practice, a margin of safety is commonly added to composition and
bioavailability of nutrients in feed ingredients, and variation in requirements caused by
environmental effects. The requirements for amino acids, fatty acids, vitamins, minerals and
energy sources were determined with diets containing purified and chemically defined
ingredients that are highly digestible to fish. Therefore, the values in the table represent near
100 percent bioavailability to the fish. This fact should be considered when formulating diets
from natural feed stuffs in which the bioavailability of the nutrients is markedly less than that
in the laboratory diets. A moderate amount of information is available for some warm water
fishes (NRC, 1993). Table 1 present the dietary nutrient levels recommenced for difference
species of fish and (Tacon, 1987) Table 2 present the dietary nutrient levels recommenced for
difference size of shrimp.
Among the major dietary nutrients required, protein is considered first because
it is the most costly among the macronutrients and cannot be substituted by other nutrients.
Protein is used very efficiently as a source of energy but for economic reason should be kept
to a minimum. Carbohydrate and fat should be used to supply most of the energy.
Feeding stuffs
Feed ingredients are selected on the basis of their nutrient composition and
available energy content as determined by chemical analyses. No single feed stuff contains
all nutrients that are needs in the correct proportion so that the nutrients in a balanced feed
formula are supplied by various feed stuffs. Each ingredient in feed formula is included
because it contributes a component necessary in the diet as shown in the proximate
composition of some ingredients present in table 3. (NRC, 1993) The composition of feed
stuffs may vary or fluctuate with season and geographic which affects feed formula. The
protein content of grains may vary from batch to batch due to differences in soil fertility,
time of harvest, genetic constitution and several other factors. The amino acid composition
of specific grains may also show wide variation. Animal protein sources are also subject to
variation as a result of manufacturing conditions and the nature of raw material Which they
are processed. Generally, the values given in composition tables are averages reflecting the
concentrations of nutrients most likely to be present.
Each feed stuff in any diet formulation should be present for a specific reason; i.e., it
is good for energy source, it is rich in a limiting amino acid, etc. In addition, each feed stuff
in a particular diet formulation should be the least costly ingredient available for its
particular function in the diet. This leads to assumption in feed formulation that same
nutrient in a particular feed stuff, such as an amino acid, is just as valuable as the same
nutrient in any other feed stuff. This allows feed formulators to interchange one feed stuff
29
Ingredients Quality Nutrient Requirements
Physical Examination Nutrient Balance
Chemical Analysis Apparent Digestibility
DIET FORMULATION
DIET PREPARATION
FEED
Pelletability Acceptability
Durability Water Stability
Sizes/ Fines Safety
Feeding/ Husbandry
Bioengineering Health
Management Breeding
Water
Feed Efficiency
Profit Fish Financial Input
QUALITY
Fig.1 Fish Nutrition and Aquaculture (Cited in Cho et al., 1985)
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
nutrition needs of fish equally well. While this assumption may not be entirely valid and
some nutritional judgement must be employed in any feed formulation, it does not seem to
be valid in most cases. As with the previously mentioned assumption, an awareness of the
potential mistake involved is necessary for the fish feed formulation so that allowances can
be made in diet formulation and problems can be anticipated and avoided.
A mixture of feed ingredients is used to meet nutritional needs. The number of
different feed stuffs used in commercial feed is small, primarily because few feed stuffs are
available that can provide the relatively high level of nutrients required by fish. The major
feed stuffs used in fish may be broadly classed as protein supplements and energy
supplements.
30
Factors in the selection of feed stuffs
Traditional feed stuff used for poultry and swine are believed to be suitable for
feeding shrimp and fish. The use of local feed stuffs however, has not been well documented
for fish and shrimp. Factors in the selection of feed stuffs are:
1. Suitable for species being cultured – knowledge of the feeding habit and nutrient
requirement of the animal are important.
2. Market price – cost can vary from one location to another. A way to consider the cost
effective is by relate cost and its nutritive value of a feed stuff as shown in this example:
Given: Local fish meal 45% Protein cost 12 Baht/ Kg
Chile Fish meal 60% Protein cost 15 Baht/ Kg
Thus: Local fish meal 12/0.45 = 26.67 Baht / Kg
Chile fish meal 15/0.60 = 25.00 Baht/ Kg
3. Available Quality and regularity of supply - with seasonal ingredients, long term storage
will become a problem.
4. Acceptability – whether it is palatable to the animal.
5. Pelletability and shelf life – high crude fiber and fat make a problem in pelletability.
Moisture should be below 10% and there should be no insect infestation. (Dy-penaflorida
1995)
Classification of feeding stuffs
Feed stuffs may be classified according to the important nutrients they contain. They
may come from conventional or non-conventional sources. Some of non-conventional
sources are also used in fish and shrimp feeds but are not yet commercially utilized as they
are limiting in availability.
1. Protein sources
Feed stuffs containing at least 20% protein are considered protein supplements.
Protein may come from animal or plant origin such as feed stuffs of animal origin have
generally high protein content ranging from 27-85% where plant sources have 20-45%.
Soybean meal, cottonseed meal, peanut meal, fish meal, meat and bone meal, blood meal,
offal meal, and cooked full fat soy bean meal have been used as protein sources in feeds for
fish. Protein quality is measured by its amino acid content. The ten essential amino acids
(EAA) required by fish and shrimp are arginine, methionine, valine, threonine, Isoluecine,
leucine, lysine, histidine, phenylalanine and tryptophan. The amino acids profiles differ
between animal and plant sources. Soybean meal provides most of the protein in
commercial fish diets. A small amount of fishmeal, a mixture of meat and bone, and blood
31
meal is typically added to fish feed to improve indispensable amino acid balance and to
increase palatability. Synthetic amino acids are not normally used in commercial fish feeds,
but recent data indicate that the use of synthetic lysine can improve protein quality of cotton
meal for catfish.
Animal protein sources are commonly marine animals while plant proteins are from
legumes and oil-bearing seeds. Legumes are valuable feed sources in the tropics because of
their abundance. Their leaves are rich in protein and minerals. The seeds are rich in lysine
though poor in methionine. Oil cake which are by products of vegetable and oil industry, are
also important. Plant protein sources may contain toxic or anti nutritive factors, but these
could be removed by soaking or treatment with heat. Iron salts and phytase
Conventional Animal and Plant protein sources

Source Characteristics
Fish meal Most common source of high- quality protein.
Shrimp head meal Good amino acids and fatty acid pattern, rich in carotenoids, high in
Shrimp meal chitin.
Squid meal Good amino acids and fatty acid pattern, rich in carotenoids.
Trash fish Good amino acids and fatty acid and fatty acids, contain cholesterol and
Meat & bone meal unidentified growth factor.
Mussel meal Nutrient composition is varied with species of fish included.
Soybean meal Contains growth promoting factors and attractants.
Copra meal Best amino acids profile but limited in sulphur amino acids.
Cottonseed meal Low in protein, abundant supply in the tropics.
Cowpea meal High in fiber, contain gossypol (toxic)
Gluten meal High in lipid
Peanut meal High protein and low fiber
Sunflower meal Good source of arginine, subject to aflatoxin contamination.
Contain no known toxins
32
Non-Conventional Animal and Plant protein sources
Sources Characteristics
Crab, Clam,Kapis, Supplies cholesterol, phospholipids, fatty acids attractant variable
Scallop, nutritional profile limited in supply.
Octopus meal Contain toxin which is removed by drying
Earthworm meal Contain uric acid
Chicken manure EAA levels close to chicken egg protein.
Milk and milk by product Rich in choline
Poultry by products High in chitin
Silkworm pupae Good amino acids and fatty acids , expensive
Brine shrimp Artemia High protein, usually cooked and dried
Golden cherry snail Rich in protein and digestible carbohydrate
Brewer’s grain High protein, good amino acids pattern low digestible require
Yeast boiling to inactive enzyme.
Low digestibility, contain mimosine , incorporation is limited (5-
Leaf meals of Ipil-Ipil 10%)
Cassava High protein content soaked or chopped and sun dried to remove
Papaya hydrocyanic acid.
Alfalfa High in protein low in fiber.
Safe for fish feed source of fat soluble vitamins.
2. Energy sources
Feed stuffs used to supply energy are defined as feed stuffs that are high in energy,
contain less than 18% fibre, and usually contain less than 20% protein. Feed stuffs
commonly used for energy in fish feeds include grains, grain milling by products and fat and
oils. Grains or grain milling by products are high in starch, which is relatively well utilized
by fish for energy. Cooking increase the digestibility of starch. Fats and oils are excellent
sources of energy and are highly digestible to fish. Feed stuffs used for energy in fish feed
are :
1. Lipid sources
Lipids or fat may also come from animals or plants animal lipids that are commonly
used in fish feeds are fish oils. Such as cod liver and sardine oil, squid liver oil, and beef
33
tallow. Plant lipid sources include corn, coconut, soybean, sunflower, peanut, cottonseed and
linseed oils.
2. Carbohydrate sources
Carbohydrate:- include starches, sugars and celluloses. They may come from cereals
or root crops. Cereals are important components in aquatic feeds. They are cheap source of
energy because of their high carbohydrate content and are also good sources of B vitamins,
especially the bran. Root crops are also excellent sources of carbohydrate but their use in
feed is limited because they are consumed by humans and cannot be digested well by aquatic
species. Carbohydrate sources include bread and wheat flour, starches (corn, potato, cassava,
and sago palm), rice bran and other rice by products, and sorghum
3. Vitamin and mineral sources

Vitamins and minerals are required in trace amounts for normal growth,
reproduction, health and general metabolism. Deficiency symptoms usually occur in
intensive system where natural food is not sufficient. Vitamins can be found listed in the
following table:
Vitamins Major natural sources

B1 (thiamine) Legumes, bran, yeast.
B2 ( Riboflavin) Yeast, liver milk, soybeans
B5 pantothenic acid Brans, yeast, animal offal fish flesh
B6 (pyridoxine) Yeast, cereal, liver
B12 (Cyanocobalamin) Fish meal, and viscera, slaughter house waste.
C (Ascorbic acid) Citrus fruits, fresh fish tissue, insects.
Niacin Yeast, legumes, forage
Biotin Liver, yeast, milk products
Folic acid Yeast, fish tissue and viscera, leaf meal
Choline Wheat germ, legumes.
Inositol Legumes, yeast, wheat germ.
A (Retinol) Fish oils.
D (Calciferol) Fish oils.
E (Tocopherol) Vegetable oils
K (Menadione) Leaf meals
34
4. Feed additives
Feed additives are substances usually added in trace amounts to a diet or feed
ingredients to preserve its nutritional characteristics; improve performance (growth and
survival) facilitate feed pelleting; facilitate feed ingestion; enhance acceptability and supply
essential nutrients in purified form. Feed additives can be classified as;
4.1 Antioxidants they are added to prevent or delay rancidity in fats and deterioration of
vitamins, pigments and amino acids. Antioxidants can be a naturally occurring
substances like vitamin E or synthetic chemicals like the following
Antioxidant Maximum allowance level

Ethoxyquin 150 mg/kg feed
BHT (butylated hydroxytouene) 0.02% of fat
BHA (butylated hydroxyanisole) 0.02% of fat
4.2. Mold Inhibitors -Feed stuffs and feeds with high moisture content (more than
12%) are susceptible to microbial attack, and consequent loss of nutritional value and
production of mycotoxin. To control deterioration, mold inhibitors are added at 0.25%
of feed. Some examples are:
Sodium or calcium propionate or its acid
Sodium, calcium or potassium sorbate or its acid
Sodium benzoate or its acid
Sodium or calcium ascorbate or its acid
Citric acid
4.3 Binders
Binders improve the water stability of feed and the efficiency of feed manufacture.
They also prevent disintegration of pellets during transport and storage. Normally, 5%
binder is used. Selection of binder is dependent upon the cost and availability;
adaptability to large scale processing techniques, desired water stability, digestibility by
the animal, reaction with other micro organisms in the intestinal tract and growing area.
The effectiveness of individual binders will depend upon; ingredient particle size,
manufacturing process and diet composition. (Heinen, 1980) Binder increase surface area
of gelatinizable material, thus increasing stability of pelleted binders may grouped as;
a. Natural sources
Marine plant extracts;-such as agar, alginates, carrageen
Plant extracts, exudates and flours;-
1. starch :-cassava starch , corn starch, sago palm starch , potato starch
2. Bread or wheat flour
Plant gums- gum arabic , guar gum, locust bean gum
Gluten- wheat gluten, corn gluten
35
3. Animal extracts;- gelatin, chitin
b. Synthetic binders – carboxymethycellulose (CMC)
4.4 Attractants or feeding stimulants
To increase feeding acceptability, the diet must have the correct appearance,
texture, density and attractiveness. The relative importance of these factors will depend
on the feeding behaviour of fish or shrimp (visual feeder or a chemosensory feeder).
Feeding stimulants are used to wean fish larvae from live food to an artificial diet or to
replace a practical fed with unconventional protein sources. Nutrient leaching can also
minimized since the feed will remain shorter in the water. Types of feed stimulants
1. Natural ingredients-squid meal, mussel flesh, shrimp, terrestrial worm, marine
fish oil, fish meal, fish solubles, fish protein hydrolysates and soybean protein
hydrolysates.
2. Purified synthetic derivatives- mixtures of L- amino acids (AA mixtures of
glycine, alanine, proline and histidine) betaine and soluble nucleotide bases.
4.5 Pigments
Pigments are added to facilitate food ingestion or impart color to fish carcass.
In the wild, the coloration is derived through ingestion of carotenoid pigments
(astaxanthin) of some invertebrates. Some animals including fish and shrimp are
believed to be unable to synthesize carotenoids de novo. Carnivorous fish can not
transform carotenoids and will deposit them in their unaltered state. Some naturally
occurring sources of carotenoids are
Astaxanthin; Crustacean meal and waste, pigmented fish oil (Capelin),
marine yeast, lobster shell cooked and boiled
Canthaxanthin. Brine shrimp nauplii (A. salina)
Xanthophyll- algae (Chlorella, spirulina sp.) Ipil- Ipil leaf meal, palm oil,
corn gluten, yellow corn, marigold petal meal ( Tagetes erects)
The natural carotenoid pigment is usually lacking in intensively farmed fish
and shrimp with no access to live food organism, thus it is necessary to fortify practical
rations with carotenoid preparation. These preparation are costly but they usually impart
same visual coloration of muscle as natural astaxanthin sources. Some synthetic
pigments available such as Asdtaxanthin, Zeaxanthin, Canthaxanthin and Lutein
4.6 Synthetic amino acids
Some protein sources are deficient in one or more amino acids, for example,
legumes and meat and bone meal are deficient in methionine. Others have imbalances
due to the presence of specific amino acid in disproportional levels, including leucine
and Isoluesine antagonism, and to a lesser extent arginine and lysine, cystein and
methionine antagonism. Results with crystalline amino acids supplementation in fish
studies, are varied. Some studies show acceptable utilization and others showed not well
utilized compare to those from intact protein. (Dy-penaflorida 1995)
36
Anti -Nutritional Factors and Toxins
Plant and some animal feed stuffs contain toxic or anti nutritive elements in
their natural state. (Liener, 1967) These harmful substances prevent the utilization of the
nutrients especially protein. Most of these toxins are removed by dry heat treatment or
cooking in water prior to feed processing.
Anti-nutritional factors and toxins Found in

Trypsin inhibitor unheated or underheated soybean.
Mimosine Ipil- Ipil leaf
Alkaloids green parts of potato plants
Goitrogens peanut, soybean.
Cyanogens cassava.
Sapogenin glycosides soybean meal, alfalfa
Cyclopropenoid fatty acids cottonseed oils
Gossypol pigment gland of cottonseed meal
Phytates sesame, soybean, peanut, cottonseed meals,
cereal hulls, germ & endosperm.
Hemagglutinins soybeans, other legumes.
Aflatoxins (Aspergillus sp.) peanut, cereals, sunflower, cottonseed, copra & cassava
Lipoxidase soybean meal
Anti vitamin D soybean meal
Anti vitamin E kidney beans
Thiaminase raw fish, herrings, mussels, clams,
Anti-biotins raw egg white
Botulin trash fish stored anaerobic
Salmonella contamination contaminated animal proteins, insects & rodent wastes
Pesticides & herbicides plant sources infected by spraying &
accumulated in fish and fish product
Heavy metals concentrated by animals and plants used as ingredient
source (e. g. mercury)
Peroxides Poorly stored and inadequately protected oils Those
with high polyunsaturated fatty acids.
37
Adverse Effect on Anti- nutritive Metabolites or Toxins
Trypsin inhibitor -Binds trypsin to form an inactive compound thus hampering proteolytic
activity of the enzyme which result to depress growth, increase size of
pancreas, decrease fat absorption and metabolizable energy. It has also
been found that inhibitor in raw soybeans cause contraction of gall
bladder, increase bile acid excretion, lowers intestinal proteolytic activity
and effects methionine metabolism.
Mimosine -A cyclic amino acid which is toxic at high levels.
It destroys hepato-pancreatic cells of shrimp.
Goitrogens inhibit the uptake of iodine by the thyroid gland thus preventing growth.
Cyanogens glycosides which release poisonous hydrocyanic acid upon hydrolysis
Gossypol reduce amino acid availability
Phytatse Binds proteins and minerals (Phosphorus, calcium, zinc, copper,
magnesium &etc.) reducing their availability
Hemagglutinins -agglutinates red blood cells. They are capable of releasing intestinal
membrane bound lipase and amylases causing these digestive enzyme to
eliminated in the faeces, thus reducing their digestive capabilities
Aflatoxin group of highly toxic and carcinogenic metabolites produced by
Aspergillus flavus, which cause liver damage.
Anti-vitamin factor forms complex with the vitamins making them unavailable
Pesticides and Chorinated hydrocarbons affect fat protein metabolism (dieldrin), and
vitamin C metabolism (Toxaphene)
Peroxides oxidize polyunsaturated fatty acids which will lead to poor growth and
survival, muscular dystrophy, de-pigmentation, and degradation of
pancreatic tissue.
In the feed composition table, it is stated whether the values are given as "as fed"
or as "received" basis (contains moisture). Sometimes it is specified that values are
given on a dry matter basis (DM). In feed formulation, as fed values are used.
"as fed" value is converted to DM as:
% DM protein = % protein as fed X 100
(100- % moisture)
DM analysis is converted to as fed basis as ;
% As fed protein = % protein DM X 100 - % moisture
100
In all cases data should always state whether they are report as "as fed" or "DM" basis
38
Fish Feed Formulation
In the past, fixed formulas were used because of the lack of sufficient nutritional
information. Presently nutritional data are available to allow the nutritionist to formulate fish
feeds on mathematics calculation or a least cost basis.
There are many method to formulate the feed for fish such as :
I. Mathematical techniques
The methods used for feed formulation are simple unless least cost diets are required.
Diets that contain few feed stuffs, or where levels of protein, energy, vitamins and minerals
are fixed, may be formulated using the Pearson square method, simultaneous equations or by
trial and error. For sample problems, feed formulation using Pearson’s square and algebraic
Equation method were given as follow:
Example 1. Diet with 2 ingredients

To formulate a diet that contains 30% protein with the following ingredients:
Fish meal 60% protein and rice bran 10% protein
1. A Pearson’s square method

1. Draw a square and place the desired protein level of the diet at the middle.
2. Place the two ingredients and their protein content at the left side of the square.
3. Subtract the protein content of each ingredient from the desired protein level of the
feed. Place the difference at the corner of the square diagonally opposite the ingredient.
( take absolute values only, that is, ignore the positive or negative signs)
4. Take the sum of the numbers at the right side of the square
5. Take the percentage of the numbers on the right (by using the sum obtained in step 4)
Fish meal 60% 20 parts
(20/50) x 100 = 40%
30
(30/50) x 100 = 60%
Rice bran 10% 30 parts
50 parts
The diet formula is thus:
Fish meal 40 g x 0.60 = 24 g protein
Rice bran 60 g x 0.10 = 6 g protein
Total 100 g = 30 g protein
1 B. Algebraic method
1. Assign a variable to each ingredient:
39
Let X = g fish meal / 100 g feed
Y = g rice bran/ 100 g feed
2. Let the first equation be for the amounts of ingredients:
X + Y = 100 ( g feed) (Eq 1)
3. Let the second equation be for the protein content:
0.60X + 0.10Y = 30 (g protein in 100) (Eq. 2)
4. Solve the two equation together. Multiply Eq. 1 by 0.1 to obtain Eq. 3:
0.1X + 0.1Y = 10 (Eq 3)
5. Subtract Eq. 3 from Eq. 2 and solve for X:
0.60x + 0.10y = 30
- (0.1X + 0.1 Y = 10)
0.5X = 20
X = 20/0.5 = 40 g fish meal
6. Substitute the value of X in Eq. 1 and solve for Y
40+ Y = 100
y = 100 - 40 = 60 g rice bran
To formulate a 30% protein diet, use 40 g fish meal and 60 g of rice bran\
Example 2. Diet with 3 or more ingredients

Formulate a diet that contains 40% protein using the following ingredients:
Fish meal 60% protein Soybean meal 40% protein
Rice bran 12% protein Wheat flour 12% protein
Assume a proportion of 2 parts fishmeal to 1 part soybean meal and a proportion of 1 part
rice bran to 1 part wheat flour.
A. Pearson’s square method

1. Draw a square and place the desired protein level of the diet at the middle
2. Group the ingredients based on protein and energy content. Calculate the weight
average protein content of each group
Protein sources
Fish meal (FM): 2 parts x 60% = 120
Soybean meal (SBM): 1 part x 45% = 45
3 parts = 165
Weight average: 165/3 = 55% protein
Energy sources
Rice bran (RB): 1 part x 12% = 12
Wheat flour (WF): 1 part x 12% = 12
40
2 parts = 24
Weight average 24/2 = 12% protein
3. At the left side of the square, place the groups of ingredients with their
corresponding weighed average protein content.
4. Proceed as in step 3 to 5 of Pearson’s square in Example 1a.
Protein Sources 55% 28
(FM& SBM) (28/43) x 100 = 65.12%
40
(15/43) x 100 = 34.88%
Energy sources 12% 15
(RB & BF) 43
5. Calculate the contribution of each ingredient within a group by multiplying the
percentage obtained in step 4 by the assumed proportion or fraction.
Protein sources
Fish meal (2 parts) = 65.12% x 2/3 = 43.41%
Soybean meal (1 part) = 65.12% x 1/3 = 21.71%
Energy sources
Rice bran (1 part) = 34.88% x ½ = 17.44%
Bread flour (1 part) = 34.88% x ½ = 17.44%
The diet formula is thus:

Fish meal 43.41g x 0.6 = 26.05 g
Soybean meal 21.41g x 0.45 = 9.77 g
Rice bran 17.44g x 0.12 = 2.09 g
Bread flour 17.44g x 0.12 = 2.09 g
100.0 g diet 40 g protein
2B. Algebraic method

1. Group the ingredients based on protein and energy content. Calculate the weighted
average protein content of each group as in step 2 of Pearson's Square above.
Weighted average from protein sources = 55%

“ “ “ “ = 12%
2. Assign variables:
Let X = g protein source/ 100 g feed Y = g energy source /100 g feed
3. Let the first equation be: X + Y = 100g feed Eq. 1.
4. Let the second equation be for the calculated protein levels of the two groups
0.55X + 0.12Y = 40 g protein Eq. 2.
41
5. Multiply Eq 1 by 0.12 to get Eq. 3.:- 0.12X + 0.12Y = 12 Eq. 3.
6. Subtract Eq 3. From Eq. 2. and solve for X:
0.55X + 0.12Y = 40
- (0.12X + 0.12Y = 12)
0.43X = 28
X = 28/0.43 = 65.12 g from protein source
7. Substitute the value of X in Eq. 1. to get value of Y:
65.12 + Y = 100
Y = 100 - 65.12 = 34.88 g from energy source
8. Calculate the contribution of each ingredient as in step 5 of Pearson‘s square.
Example 3. Diet with some ingredient in fixed percentages

Formulate the diet containing 40% protein with fish meal (60% protein) and copra
meal (20% protein). The diet will also contain for every 100 g:
wheat flour (12% protein) 15 g
Mineral mix 2g
Vitamin mix 2g
2 A. Pearson ‘s square method
1. Calculate the total weight of fish meal and copra meal that should be in the diet.
100 – 19 = 81 g / 100 g of feed
2. Calculate the amount of protein contributed by bread flour and subtract it from the total
amount of protein required. (Note that the mineral and vitamin mix do not contain protein).
15 g x 0.12 = 1.8 g protein from wheat flour
40 g protein - 1.8 = 38.2 g protein from fish meal and copra meal
3. Calculate the percent protein from the total weight contributed by fish meal and copra
meal (38.2/81) x 100 = 47.16% protein
Proceed with the steps of Pearson’s square method as in Example 1a.
Fish meal 60% 27.16
(27.16/40) x 81 = 55%
47.16
(12.84/40 x 81 = 26%
Copra meal 20% 12.84
40
The diet formula is thus: Fish meal 55 g x 0.60 = 33 g protein

Copra meal 26 g x 0.20 = 5.2 g protein
Wheat flour 15 g x 0.12 = 1.8 g protein
Mineral mix 2g
Vitamin mix 2g
42
Total 100 g 40 g protein
3 B. Algebraic Method
1. Calculate the total weight of fish meal and copra meal that should be present in diet.
100 – 19 = 81 g / 100 g feed
2. Calculate the amount of protein contributed by wheat flour and subtract it from the total
amount of protein required.
40 g – (15 x 0.12) = 38.2 g protein from fish meal and copra meal
3. Assign variations: Let X = g fish meal and Y = g copra meal
4. Let the first equation be:
X + Y = 81 Eq. 1.
5. Let the second equation be for the amount of protein:
0.60X + 0.2Y = 38.2 Eq. 2.
6. Multiply Eq 1 by 0.20 to form Eq. 3.
0.2X + 0.2Y = 16.2 Eq. 3.
7. Subtract Eq. 3 from Eq. 2 and solve for X :
0.60X + 0.2Y = 38.2 Eq. 2.
- (0.2X + 0.2Y = 16.2) Eq. 3.
0.40x = 22
x = 22/0.40 = 55 g fish meal
8. Substitute value of X in Eq 1 and Solve for Y:
55 + Y = 81
Y = 81 - 55 = 26 g copra meal
3. Use a worksheet
1. Prepare a work sheet like Worksheet 1. Fill in the require ingredients and the
corresponding amounts (columns 1 and 2.)
2. Fill in the given nutrient content based on proximate composition of each ingredient (insert
boxes of columns 3-7, worksheet 2)
3. Calculate the amounts of nutrients contributed by the ingredients (columns 3-7). For
example, 17g of fish meal with 72% protein contribute 12.2 g protein to the diet. To fill up
the columns, multiply the amount of ingredients (column2) and the % nutrient content (insert
boxes) as shown in Worksheet 2.
4. Calculate the contribution of soybean meal and rice bran by subtracting the known protein
contributions of the other ingredients from the total protein requirement of 35 g.
35 g protein - (12.2 + 2.4 + 0.7) = 19.7 g protein from soybean meal and rice bran
This 19.7g protein in 59 g of the two ingredients means a protein content of 33.4%
43
5. Use Pearson‘s square method to calculate the amounts of soybean meal and rice bran to be
added to the diet.
Soybean meal 42% 23.4
(23.4/32 X 59 = 43.2)
33.4
(8.6/32) X 59 = 15.8 g
Rice bran 10% 8.6
32
Fill in the rows for soybean meal and rice bran in Worksheet 3.
Determine the amounts of the remaining feed stuffs and the nutrients they supply.
Take lipids – the contributions of the five ingredients total 7.6. Since the total fat required is
9%, add 1.4% cod liver oil to fulfill the fat requirement. Now, the total amount of ingredients
is 99.4 g. To make up 100 g diet, add 0.6 g filler (Worksheet 3)
Compute the total cost of the feed (excluding processing) given the prices below:
Ingredient Cost (B/Kg) Cost (B/ 100 g diet)
Fish meal 18.00 17.0 x 0.018 = 0.31
Copra meal 10.00 14.0 x 0.010 = 0.14
Rice bran 12.00 43.2 x 0.012 = 0.52
Bread flour 6.00 15.8 x 0.006 = 0.09
Vit /Min Mix 300.00 3.0 x 0.300 = 0.90
Cod liver oil 100.00 1.80 x 0.100 = 0.18
Filler 20.00 0.60 x 0.020 = 0.012
2.212
This means that the total feed cost (excluding processing) is 22.12 Baht / Kilogram.
Compute the total energy contribution of protein, fat and carbohydrates (NFE) .
Protein: 35g x 4 Kcal/g = 140 Kcal
Fat: 9g x 9 Kcal/g = 81 Kcal
NFE: 33.8g x 4 Kcal/g = 135 Kcal
356 Kcal
This amount of energy satisfies the requirement in the sample problem.
In the worksheets of example 4, only the macro-nutrients (protein, fat. carbohydrate)
were balanced as specified in the problem. After the diet formula has been established,
the essential amino acids can be computed and balanced as well. In finalizing the feed
formulation, additives (attractants, anti-oxidants and mould inhibitors) are added to
improve diet palatability or acceptability, and to lengthen the shelf life. Some
adjustments are done to reached a final balanced formula. (Dy-penaflorida 1995)
II. Least cost formulation
44
The least cost formulation is the computerized process of evaluating all
ingredients, their nutrient levels and their costs so that when the diets are prepared, they
provide the best nutrient levels at the least cot. The emphasis is on providing optimal
nutrient levels and low cost. The formulation process is actually conducted using a
series of linear programming techniques. As the solution to the equation is solved, the
computer automatically adjusts the most expensive ingredient used and reformulates.
The computer continues reformulating until the most economical dietary formulation is
attained. If a feasible solution is not attainable the formulation process halts and a
statement to the effect is issued. The use of least cost computer program to formulate
feeds, the following information is needed.
1. Cost of feed ingredients and nutrient concentrations in feed stuffs
2. Nutrients requirement
3. Nutrients availability from feed stuff
4. Nutritional and non nutritional restrictions
The primary constraint limiting the use of least cost programs for formulating fish
feeds is that relatively few feed stuffs are available that suitable for use in fish feeds.
Many feed ingredients are unsuitable for use in fish feed because of their poor
nutritional content or because of manufacturing constrains. Nutrient levels
recommended for practical fish feeds are given in table
Several constraints that inhibit the widespread use of least cost formulation of
fish feeds in addition to the lack of sufficient number of suitable feed stuffs. It
include a lack of knowledge of the nutrient levels that results in maximum profit. It
opposed to levels that maximize weight gain. Fish feed mills do not have the capacity
to store a large number of different ingredients, and the logistics of obtaining a wide
assortment of feed stuffs on a timely basis. A limited form of least cost feed
formulation is used to formulate fish feeds. Cottonseed meal, milo and meat and bone
meal are often used to replace a part of soybean meal, corn and fish meal, respectively
depending on the cost.
One has to keep in mine that the process of least cost formulation is only as
good or as efficient as the nutritionist using it. Poor quality diets can be made with even
the best formulation programs if the nutritionist does not know how to use it. Every
diet must be evaluated carefully to assure that all nutrients are provided. Just because a
computer calculated the ration don't assume that it is adequate for the fish. Computer
formulation is an excellent method to evaluate the nutritive value of feed ingredients.
When formulating rations, the primary emphasis is always stressed upon providing the
optimum nutrients rather than reducing the feed cost. Most of the least cost formulation
report shadow price, it is the value of ingredient/unit which is change due to market
price and it indicate how much of these ingredient worth in the formula per unit price.
45
Feed manufacture
General material processing is any procedure undertaken to change the
physical characteristics of an ingredient to improve its blending ability or to increase its
nutrient availability. Feed manufacture involves grinding, mixing, agglomeration and
forming feed ingredients into homogenous pellets that are water stable. Feeds for fry or
small fingerlings are usually fed finely ground meal-type or pellet feeds that have been
crumbled to reduce particle size. The bulk of feeds used in the fish industry are
manufacture by pelleting or extrusion into floating pellets.
General scheme of feed manufacture is the same, regardless of the feed type,
floating or sinking pellets.
1. Size reduction procedures

1. Cutting or chopping
2. Crushing – roller mill application of compressive force
a. relatively uniform product- vary little pulverized material
b. If previously steam treated, the grain will have a flaked appearance.
Usually referred to as flaked or crimped ex. Ball mills, percussion mills, jaw crushers
3. shearing – combination of cutting and crushing
4. grinding - hammer mills , jet mills, centrifugal impact mills, attrition mills
Grinding Equipment – General impact grinding- particles to be reduced meet the
milling surface at a high velocity. Reduction rate depends on the velocity of the grain.
Type of grinding equipments are;
1. Jet mill material is blown into an impact plate at speed of 27,000 to 36,000
ft/min. Efficiency depend upon velocity, quality of material being reduced, quality and
conformation of the impact surface
2. Centrifugal impact mill – two parallel horizontal disk with rings of hard
steel impactors at the circumference. Used in the flour milling industry due to its fine
grinding capabilities.
3. Hammer mill most common grinding tool. It is versatility to produce a fine
grind from a greater variety of raw materials than any other machine. Rotor with steel
bars connected that beat grain into a screen. It consists of stationary or swing hammers
mounted on a rotor that is affixed to a rotating shaft. Shaft speed are 1,200-4,000 rpm.
dynamically and statically balance to avoid harmful vibration Peripheral speeds (speed
that hit the material) are 5,000 - 27,000 ft/min. It must be
4. Attrition mill consists of two plates or discs equipped with replaceable
wearing surfaces. This mill action by combination of cutting, crushing, shearing and
impact. Two plates or disc equipped with replaceable wearing surfaces. One disc
rotated (burro mill)- shredding, defiberizing, coarse reduction of easily ground
materials, blending of materials that are difficult to blend. Two disc rotated (double
46
runner mill or heavy duty plate mill or fine grinding mill) closeness of two plates, plate
types, rate of feed, plate wear, type of product and plate speed.
2. Sieving/Screening and grading

It is necessary to screen the ground feed material. A number 40 mesh size (425 um) is
usually used. Coarse materials that do not pass through the sieve are returned back to the
grinder and reground until all passes through the sieve. The screen action is accomplished by
mounting the screen or a number of screens (one over the other) on a mechanically vibrating
frame. The screens are normal mounted on this frame at a slightly downward slope to allow
the retained coarse material to discharge at the lower end of the screen sheet. For multi-
decked vibrating screens, the screen with the larger perforations is mounted on the top layer,
the subsequent screens with progressively smaller openings. In some feed mills, the hammer
mill is mounted just on top of the vibrating screen to facilitate material handing from the
hammer mill to the grading screen.
3.. Weighing
It is vary important step in feed preparation. The feed components as prescribed in
feed formulation must be weighed as accurately as possible. A good and accurate balance
with a tarring device is best as it can be adjusted to zero and allow weighing with the
container or weighing of components within the container.
4. Mixing of feed components
Most mixing of the feed components to form the meal is done in batch using
suitable type mixer. Mixing may be defined as the scattering of dissimilar parts into a blend.
Unless already premixed, liquids should be added after all dry ingredients have been mixed.
Total mixing time is critical and is influenced by the composition of the formula. Three
mechanisms are involved in the mixing process.
1. Transfer of groups of adjacent particles from one location in the mass to
another.
2. Diffusion: distribution of particles over a freshly developed surface
3. Shearing : slipping of particles between others in the mass
There are three general types of mixer used in the feed milling industry like
1. Horizontal ribbon type mixer (batch and continuous)
2. Vertical boot loading mixer
3. Oscillating screw mixer
Normally mixing is done through the horizontal ribbon type mixer, especially when
mixing ingredients of different particulate size with some liquid. The design and
configuration of mixing ribbon is essential in relation to the kind of material being mixed.
Mixing particularly when working with dry powders in the absence of dosing liquids can
cause fine feed particles to escape from the mixer unless proper covers are provided for the
mixing vessel. The power and speed of the mixer drives should be designed to meet the
47
needs of the mixer at maximum loading and material density. Mixer should be constructed
for easy loading, meal discharge as well as cleaning of mixer until every mixing operation,
since residue meal can cause quality problems in the subsequent mixing batch or load.
5. Meal conditioning
After feed ingredients are mixed into meal, it will require conditioning prior to
pelleting. Conditioning of the ground feed mixture (mash) is the most critical stage in the
manufacture of shrimp feeds. Conditioning is a combination of the introduction of certain
liquids, kneading of the meal and heat treatment to allow some of the starch and other binders
to gel. The meal is subjected to low temperature steaming of about 1-2 Kg/cm2 in enough
time so as to allow the starch material to form gel. Wet steam with condensation will cause
mesh clumping will choke the die and roller assembly and / or produce uneven colored
pellets. Saturated steam would be preferred and control of steam temperature is vary essential
in order not to affect some of the heat sensitive ingredients in the feeds. Steam is normally
added to the feed mixture to increase the moisture content to approximately 16-18% and
temperature to 80-90 c before entering the pellet die. Most pellet mills are equipped with one
to three direct or indirect steam conditioner barrels where steam can be injected directly into
the mesh or into the conditioner jackets to partially gelatinize starch and/or active binders
which aids in binding of ingredients. The jackets maintain higher temperatures within the
conditioning barrel. Under the conventional conditioning and pelleting process, about 30-
35% of starch is gelatinized. Depending on the amount of starch in the formula and type of
ingredients used. This level of starch gelatinization may not be sufficient to produce a good
water stable pellet. It is recommend that a pelleted shrimp feed should achieve at least 50%
starch gelatinization.
Increasing the retention time or use of multiple conditioners and control of steam
quality can raise starch gelatinization. Under these conditions, the feed mixture in the third
conditioner should reach the pellet a temperature of more than 90 c before entering the pellet
die.
6. Pelleting and Extrusion

Pelleting is a process of transforming soft feed mixture into hard pellets by forcing it to pass
through holes in a metal die plate which are then cut to a pre determination size. The
transformation of a soft, often dusty feed into a hard pellet is accomplished with compression,
extrusion and adhesion. By the nature of its application, shrimp feeds should be in solid form
that will not readily disintegrate upon contact with water. Since shrimp are slow eaters and
bottom dwellers, the water stability of shrimp feeds is important in minimizing the leaching
of nutrients from the feed at the same time reduces the contamination/pollution of pond water
cause by unconsummated and dissolved feeds.
48
7. Pellet cooling and drying
Vertical cooler drier is commonly used for small capacity feed plants in
conjunction with the pellet crumbing. Pellets are discharged from the mill into the top of a
flat sided hopper and grouped into an attached cooling bin pellets leave the bin at the bottom
via discharge gate at a rate regulated by the amount of pellets entering the cooler.
Horizontal cooler and dryer, this type of cooler uses a number of horizontal
moving perforated trays carrying the hot pellets counter current to the cooler air. The vertical
cooler, feed is discharged into a dust collecting system which removes the fines or particles
that separates from the pellets.
8. Pellet Crumble
The primary grains are ground through a hammer mill upon receiving. The
formulation is weighed, mixed, and then reground. After regrind, the mixed feed ingredients
are ether pelleted or extruded. The pellets are dried, crumbled, screened, fat coated and
bagged or stored in bulk.
References
Dy-penaflorida, Veronica. 1995 Lecture notes on fish nutrition. Ingredients for feed
manufacture . Aquaculture Department , Southeast Asian fisheries Development
Center. Training and information Division . Tigbauan, Iloilo, Philippines
Heinen, JM. 1980. Evaluation of some binding agents for crustacean diets. Prog. Fish Cult.
43:142-145.
Jiener, IE. 1967. Toxic factors in protein foods. Bailey L. (ed.) Techniques in protein
chemistry. 481-500 Elsevier Pub. Co., Amsterdam.
National research council 1993. Nutrient Requirements of fish. National Academy Press .
Washington, D.C.
Robinson, E. H. 1991. A practical guide to Nutrition, Feeds, and Feeding of catfish. Delta
Branch experiment station , Stoneville, Mississippi. MAFES Bulletin 979.
Tacon Albert G.J. 1987 The nutrition and feeding of farmed fish and shrimp - A training
manual 1. The essential nutrients. A report prepared for FAO trust Fund
GCP/RLA/075/ITA project. Support to the regional Aquaculture Activities for Latin
America and the Caribbean.
49
TABLE 1 Nutrient Requirements for Channel Catfish, Rainbow Trout, Pacific Salmon, Common Carp, and
Tilapia as Percentages of Diet, Milligrams per Kilogram of Diet, or International Units(IU) per Kilogram of Diet(as-
fed basis)
Channel Rainbow Pacific Common Tilapia
Catfish Trout Salmon Carp
Energy Base
(kcal DE/kg diet) 3,000 3,600 3,600 3,200 3,000
Protein, crude(digestible),% 32(28) 38(34) 38(34) 35(30.5) 32(28)
Amino acids
Arginine, % 1.20 1.5 2.04 1.31 1.18
Histidine, % 0.42 0.7 0.61 0.64 0.48
Isoleucine, % 0.73 0.9 0.75 0.76 0.87
Leucine, % 0.98 1.4 1.33 1.00 0.95
Lysine,% 1.43 1.8 1.70 1.74 1.43
Methionine + cystine, % 0.64 1.0 1.36 0.94 0.90
Phenylalanine + tyrosine, % 1.40 1.8 1.73 1.98 1.55
Threonine,% 0.56 0.8 0.75 1.19 1.05
Trytophan, % 0.14 0.2 0.17 0.24 0.28
Valine, % 0.84 1.2 1.09 1.10 0.78
n-3 fatty acids, % 0.5 - 1 1 1-2 1
n-6 fatty acids, % - 1 - 1 0.5 - 1
Macrominerals
Calcium, % R IE NT NT R
Chlorine, % R 0.9E NT NT NT
Magnesium, % 0.04 0.05 NT 0.05 0.06
Phosphorus, % 0.45 0.6 0.6 0.6 0.5
Potassium, % R 0.7 0.8 NT NT
Sodium, % R 0.6E NT NT NT
Microminerals
Copper, mg/Kg 5 3 NT 3 R
Iodine, mg/Kg 1.1E 1.1 0.6 - 1.1 NT NT
Iron, mg/Kg 30 60 NT 150 NT
Manganese, Mg/Kg 2.4 13 R 13 R
Zinc, mg/Kg 20 30 R 30 20
Selenium, mg/Kg 0.25 0.3 R NT NT
Fat soluble vitamins
A, IU/Kg 1,000 -2,000 2,500 2,500 4,000 NT
D, IU/Kg 500 2,400 NT NT NT
TABLE1 (continued)
E, IU/Kg 50 50 50 100 50
K, IU/Kg R R R NT NT
Water Soluble vitamins
Riboflavin, mg/Kg 9 4 7 7 6
Pantothenic acid, mg/Kg 15 20 20 30 10
Niacin, mg/Kg 14 10 R 28 NT
Vitamin B12mg/Kg R 0.01E R NR NR
Choline, Mg/Kg 400 1,000 800 500 NT
Biotin, mg/Kg R 0.15 R 1 NT
Folate, mg/kg 1.5 1.0 2 NR NT
Thiamin, mg/Kg 1 1 R 0.5 NT
Vitamin B6 mg/Kg 3 3 6 6 NT
Myoinositol, mg/Kg NR 300 300 440 NT
Vitamin C, mg/Kg 25-50 50 50 R 50
50
TABLE 2 Recommended dietary nutrient levels for carnivorous shrimp species 1/
Shrimp size class 2/
Nutrient level Larval PL1-25 PL 25-1g Juvenile Grower
Crude lipid. % min 14 13 12 11 10
Marine : plant lipid 3/ 6:1 6:1 6:1 6:1 6:1
Cholesterol, % min 4/ 2 1.5 1.5 1 1
Crude Protein, % min 65 62 59 55 53
Amino acids, % min 5/
Arginine 3.53 3.36 3.20 2.98 2.87
Histidine 1.00 0.95 0.91 0.85 0.82
Isoleucine 1.55 1.48 1.40 1.31 1.26
Leucine 3.18 3.04 2.89 2.69 2.60
Lysine 3.34 3.19 3.03 2.83 2.73
Methionine 1.23 1.17 1.11 1.04 1.00
Cystine 0.61 0.59 0.56 0.52 0.50
Phenylalanine 1.75 1.67 1.59 1.48 1.43
Tyrosine 1.77 1.69 1.61 1.50 1.45
Threonine 2.18 2.08 1.98 1.85 1.78
Trytophan 0.61 0.59 0.56 0.52 0.50
Valine 1.93 1.84 1.75 1.64 1.58
Carbohydrate, % max 5 10 15 15 20
Crude fiber, % max 6/ 1 1 1.5 1.5 2
Major minerals, %
Calcium, % max 3 3 2.5 2.5 2
Available phosphorus, % min 1.8 1.6 1.4 1.2 1.2
Potassium, % min 1.1 1.0 0.9 0.10 0.08
Magnesium, % min 0.18 0.15 0.13 0.10 0.08
Trace minerals, mg/kg min
Iron 100 90 80 70 60
Zine 120 110 100 90 80
Manganese 60 55 50 45 40
Copper 12 11 10 9 8
Cobalt 1.2 1.1 1.0 0.9 0.8
Iodine 6 5.5 5 4.5 4
Chromium 1.0 0.9 0.8 0.7 0.6
Selenium 0.25 0.23 0.21 0.19 0.17
Vitamin A IU/kg min 7/ 6000 5500 5000 4500 4000
51
TABLE 2 (continued.)
Shrimp size class 2/

Nutrient level Larval PL1-25 PL 25-1g Juvenile Grower
Vitamin D3 , IU/kg min 7/ 2000 1800 1600 1400 1200
Vitamin E (mg/kg min) 200 180 160 140 120
Vitamin K (mg/kg min) 12 11 10 9 8
Thiamine (mg/kg min) 30 28 26 24 22
Riboflavin (mg/kg min) 30 28 26 24 22
Pyridoxine (mg/kg min) 30 28 26 24 22
Pantothenic acid(mg/kg min) 75 70 65 60 55
Nicotinic acid(mg/kg min) 150 140 130 120 110
Biotin (mg/kg min) 0.25 0.23 0.21 0.19 0.17
Folic acid (mg/kg min) 6 5.5 5 4.5 4
Vitamin B12 (mg/kg min) 0.04 0.037 0.034 0.031 0.028
Vitamin C (mg/kg min) 500 450 400 350 300
Choline (mg/kg min) 1600 1500 1400 1300 1200
Inositol (mg/kg min) 700 650 600 550 500
1/ Dietary nutrient levels recommended for clear-water 'intensive' aquaculture systems (ie. Tanks, cages and
raceways)
2/ Shrimp size class: larval-protozoea substage 1 to post-larval substage 1 (PLI); PLI to PL25 (25 days from
PLI); Pl25 to lg;
juvenile- 1g to 10 g; grower-10g to harvest; broodstock-10g+
3/ marine lipid includes shrimp head oil, marine fish body oil, marine fish liver oil, or marine invertebrate oils.
So as to satisfy a
possible dietary phospholipid requirement, a concentrated source of phospholipid should be added in the
form of soybean oil
or a soy-lecithin preparation
4/ Cholesterol can be added eirther in purified form or by using dietary lipid sources naturally rich in cholesterol
such as shirmp
head oil.
5/ Amino acid requirement based on the carcass essential amino acid patten of the short-necked clam (Table
4)
6/ Maximum limit refers only to crude fibre of plant origin, and excludes crude fibre derived from shrimp meal
(ie. Chitin)
7/ Suggested minimum dietary vitamin levels required to prevent deficiency signs. Values in parentheses
indicate suggested
dietary vitamin levels talking into account processing, storage and leaching losses; the latter being 2 to 5
times greatet than
the recommended dietary requirements due to the extremely slow and extended feeding habits of marine
shrimp, and to
compensate for the considerable losses of vitamins which occur through leaching, However, the actual
multiplication factor
used should be adjusted in situ (ie. Either higher or lower) depending on the water stability of the diet and
the feeding response
of the shrimp to the diet, and finally on the time period the feed remains in the water before total
consumption.
52
TABLE 3 Proximate Composition and Digestible and Metabolizable Energy Values for Natural and Chemically Defined Ingredients
Commonly Used in Fish Feeds (as-fed basis)
Metabolizable
Digestible Energy Energy Proximate composition
Typical (kcal/kg) (kcal/kg) (%)
Dry
Ingredient International Matter Rainbow Channel Chinook Rainbow Crude Crude Crude
Feed Number (%) Trout Catfish Salmon Trout Protein Fat Fibre Ash
Alfalfa meal, dehydrated, 17 percent protein 1-00-023 92 513 730 - 859 17.1 2.8
24.1 9.8
Blood meal, spray dehydrated 5-00-381 93 4,289 3,410 1,864 3,410 89.2 0.74 1.0 2.3
Brewers grains, dehydrated 5-02-141 92 - - - - 23.1 6.4 13.7 3.7
Canola meal, prepress solvent extracted 5-06-145 93 2,725 - 2,373 - 38.0 3.8
11.1 6.8
Corn distillers grains with solubles, 5-28-236 91 - - - - 27.0 9.3 9.1 6.4
dehydrated
Corn distillers solubles, dehydrated 5-28-237 90 2,265 - - 2,283 27.6 8.5 4.6 7.5
Corn gluten meal, 60 percent 5-28-242 91 4,260 - - 3,554 60.4 1.8 1.5 2.1
Corn 4-02-935 88 - 2,200 - - 8.5 3.6 2.3 1.3
Corn, extrusion cooked 4-02-935 88 - 3,060 - - 8.5 3.6 2.3 1.3
Cotton seed meal, sovent extracted 5-01-619 92 2,474 3,110 - 2,468 41.7 1.8 11.3 6.4
Crab meal, process residue 5-01-663 92 - - - 3,214 32.0 2.5 10.6 41.0
TABLE 3 (continued)
Metabolizable
Dry
Fish solubles, condrated 5-01-969 50 - - - - 31.5 6.1 0.5 9.6

Fish solubles, dehydrated 5-01-971 93 3,426 - - 3,345 64.3 8.2 1.3 2.5
Fishmeal, anchovy, mechanically 5-01-985 92 4,204 - 4,828 4,328 65.4 7.6 1.0 14.3
extracted
Fishmeal, catfish by-product, 5-09-835 92 - - - - 50.8 9.6 0.5 18.0
mechanically extracted
Fishmeal, herring 5-02-000 92 4,340 - 4,876 - 72.0 8.4 0.6 10.4
Fishmeal, menhaden 5-02-009 92 - 4,060 4,063 - 64.5 9.6 0.7 19.0
Fishmeal, tuna 5-02-023 93 - - - - 59.9 6.8 0.8 21.9
54
TABLE 3 (continued)
Metabolizable
Dry
Fishmeal, white 5-02-025 92 3,211 - - 2,974 62.3 5.0 0.5 21.3

Meat meal 5-09-323 93 - - - - 55.6 8.7 2.3 27.0
Meat and bone meal 5-09-322 94 3,186 2,930 - - 50.9 9.7 2.4 29.2
Molasses, sugar cane, dehydrated 4-04-695 94 - - - - 9.6 0.8 6.2 12.5
Peanut meal, solvent extracted 5-03-650 92 - 3,370 - - 49.0 1.3 9.9 5.9
Poultry by-product meal 5-03-798 93 3,459 - 3,633 2,982 59.7 13.6 2.1 14.5
Poultry feather meal 5-03-795 93 - 3,400 3,250 - 83.3 5.4 1.2 2.9
Rice bran, with polishings 4-03-928 91 - 2,110 - - 12.8 13.7 11.1 11.6
Rice bran, with germ 4-03-930 91 - - - - 14.0 1.5 12.9 10.8
Solvent extracted
Rice polishings 4-03-943 90 - - - - 12.8 14.6 5.3 7.4
Shrimp meal, process residue 5-04-226 88 - - - - 39.5 3.2 12.8 27.2
55
TABLE 3 (continued)
Metabolizable
Dry
Sorghum (milo) 4-04-444 89 - - - - 9.9 2.8 2.3 1.8

Soybean seed, steam cooked, full fat 5-04-597 90 3,809 - - 3,641 38.0 18.0 5.0 4.5
Soybean meal, solvent extracted 5-04-604 90 - 3,010 - - 44.0 1.1 7.3 6.3
Soybean meal, solvent 5-04-612 - 2,934 - 3,250 2,566 48.5 0.9 3.4 5.8
Extracted without hulls
Sunflower meal, solvent extracted 5-04-739 93 - - - - 45.5 2.9 11.7 7.5
Wheat 4-05-268 88 - 2,400 - - 12.9 1.7 2.5 1.6
Wheat bran 4-05-190 89 - 2,790 - - 16.4 4.0 9.9 5.3
Wheat flour 4-05-199 88 1,588 - - - 11.7 1.2 1.3 0.4
Wheat middlings 4-05-205 89 2,173 - 2,032 2,237 17.0 4.3 8.0 4.6
Yeast, brewers, dehydrated 7-05-527 93 3,522 - - 2,922 42.6 1.0 3.2 6.6
Yeast, torula, dehydrated 7-05-534 93 3,421 - - - 49.0 1.5 2.2 7.7
Casein a 5-01-162 91 - 4,400 - - 84.3 0.6 Trace 2.1
56
TABLE 3 (continued)
Metabolizable
Dry
Cellulose powder a b 96 - - - - 0 0 92.6 0

Corn starch a b 88 - 2,700 - - 0.2 Trace 0.08 0.08
Corn strach, cooked a b 88 - - - - 0.2 Trace 0.08 0.08
20 percent of diet - 3,400 - - - - - -
40 percent of diet - 2,800 - - - - - -
Dextrin a 4-08-023 90
30 percent of diet - 2,920 - - - - - -
60 percent of diet - 1,920 - - - - - -
Gelatin a 5-14-503 90 - 4,700 - - 87.6 0.1 Trace Trace
NOTE : Dash indicates data were not available.

a Purified ingredients
b An International Feed number is not assigned
57

Aquaculture of Indigenous Mekong Fish SP

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Aquaculture of Indigenous Mekong Fish SP

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Component Report No.

Aquaculture of Indigenous Mekong Fish Species

Training in Feeds and Feeding for Indigenous Mekong Fish Species

Amonrat Sermwatanakul, Nantiya Unprasert Mali Boonyaratpalin and

Bangkok, May 2004

Outline of the Course and Schedule

Nutrient requirements were described with respect to protein, fat, carbohydrate,

Feed formulation part of the program included introduction to nutrient

Proteins are composed of amino acids which may be described as essential

Carbohydrates are found in greater abundance primarily in plants and

Results of the Workshop

• Simple approaches to assessing nutritional requirements

• The basics of natural food production in ponds

• The above training carried out

• A report to the Coordinator,

• A presentation (e.g. PowerPoint) covering the main outcomes of the training

• Participants presented their ongoing or planned research on feed and feeding

• Prepare the final report as described under 'Outputs'

Implementation of the training event

The Component provided logistical support as well as secretarial support, office

Activity Days Start Finish

Report preparation 27 February 2004

1 Introduction to fish nutrition 3.5

Lectures and discussion= 21 hrs., Practicum 6.5 hrs.

Number of participants: 15-20

Name Qualification Affiliation

Tuesday, February 24, 2004

08:30-12:00 Introduction to Fish Nutrition

13.00-16.00 Fish Feeds

16.00-16.30 Discussion Period

Wednesday, February 25, 2004

08:30-12:00 Feeding Fish

14:00-16:00 Feeding Systems and Feed Management

16.00-16.30 Discussion Period

08:30-12:00 Fish Feed Formulation Practice

13:00-16:30 Open Discussion

Friday, February 27, 2004

08:30-12:00 Formulating your own feed (lecture)

13:00-16:00 Fish Feed Formulation Practice (continued from Feb 26)

16.00-16.30 Discussion Period

Breaks: 10.00-10.15 and 14.30-14.45

4. Have you ever been involved in fish nutrition?

Cambodia Loa PDR Thailand Vietnam

Area of Job Researcher (2) Fish Breeder Research

Each group prepared their own farm made feed as follows.

Country: Udonthani, Thailand

Title: Dietary Requirement of Cirrhinus microlepis Broodstock

Experimental unit = 9 pens in a 3200 m2 earthen pond

Experimental design = CRD with 3 treatments and 3 replications

Tuesday, February 24, 2004

08:30-12:00 Introduction to Fish Nutrition

13.00-16.00 Fish Feeds

16.00-16.30 Discussion Period

Wednesday, February 25, 2004

08:30-12:00 Feeding Fish

14:00-16:00 Feeding Systems and Feed Management

16.00-16.30 Discussion Period

08:30-12:00 Fish Feed Formulation Practice

13:00-16:30 Open Discussion

Friday, February 27, 2004

08:30-12:00 Formulating your own feed (lecture)

13:00-16:00 Fish Feed Formulation Practice (continued from Feb 26)

16.00-16.30 Discussion Period