Ingredient selection and Feed Formulation for Fish and Shrimp

Prof. Kim, Jeong-Dae Ph.D.Aquafeed Nutritionist

Kangwon National UniversityChuncheon 24341, Korea

Talk on Nov. 26, 2019University of Brawijaya, Indonesia

Good Ingredients for Better Feed

38.1kg

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Requirements?

Nutrients

Manufacturing processFeed handling/storage

Customer Input/Feedback

Feeding Practices-Quantity and Frequency

Feed design-Formulation system

Nutrient Management Process영양관리 순서도

Ingredients

1.소비자요구 2.요구량측정

3.연구/원료/설계

4.생산5.운송

6.현장적용

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Feed Design

• Nutrients• Ingredients• Non-nutritive Ingredients

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Feed Design

• Nutrients:- Requirements (요구량)- Availability (이용성)- Interactions (상호관계)

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• Ingredients:- Physical properties (물리적특성) - Nutrient content (영양성분)- Nutrient availability (영양이용성)- Anti-nutrients (반영양인자)- Attractability/Palatability(기호성)- Storage (저장)- Color (색깔)

• Non-nutritive Ingredients:- Binders (점착제)- Pigments (착색제)- Antioxidants (항산화제)- Mold inhibitors (항곰팡이제)

Feed Selection

• Feed type – nutrition level of growing stage• Feed Form (floating/sinking)• Feed size

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Feed Selection: Broodstock and Larvae

• Not designed for rapid growth• Higher energy • Must supply nutrients needed for

gonad development and early larval stages

• Fatty acids• Vitamins• Pigments

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• Some species, especially marine or carnivorous, cannot move directly to feeds (먹이생물)

• Require live foods or highly specialized larval feeds (전문사료)

• Micro-feeds• Liquid feeds

Broodstock Feeds Larval Feeds

Feed Selection: Starter & Growout

• High protein/energy (고단백질/에너지)• Particle sizes < 2mm (사이즈 2mm미만)• High attractability/palatability(기호성)• High digestiblity(소화율)

• Poorly developed digestive system• Fortified with micronutrients(미량원소)• Immune?(면역?)

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• Majority of feed volume and cost(사료량과가격)

• Reduced nutrient density(적정영양수준)

• Focus on efficient growth(사료효율)

Starter Feeds Growout Feeds

Feed Selection – Finisher and Specialty

• Similar to growout feeds(사료량,가격)• Lower nutrient density(적정영양수준)• Prepare fish for harvest (수확감안)

• Opportunity to impact final product(최종생산물감안)

• Body composition (체성분)• Pigmentation (착색)• Shelf-life (유통기한)• Off-flavor (해감내)

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• Medicated (약제)• Immune (면역)• Low-pollution (저오염)• Recirculating system (순환여과식)• Organic (유기농)

Finisher Feeds Specialty Feeds

Feed Selection – Form

• Microparticulate (미립자)• Mash (분말)• Crumbles (크럼블)• Pellets (펠렛)• Extruded pellets (익스트루젼-압출성형)

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Feed Selection – Size

Maximum particle size is limited by size of mouth opening

사료최대크기는 입 벌린어류 입크기로 판단

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Fish meal/oilCereal

by-product meals & oils

Oilseed meals,

Pulses & Oils

Novel feed ingredients

Land animal protein meals

and fats

40essential nutrients

Nutrient sources from agricultural farming activity

Which ingredients can be used for aquafeed production?

Nutrient sources from fishing activity

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Feed safety

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Feed Ingredients and Physical Characteristics

Formulating Feeds

• Nutritionally complete feeds should be used whenever natural foods are absent or where natural foods only make a small contribution to nutrition

• When substantial amounts of natural productivity are available, supplemental feeds don’t need to contain all essential nutrients

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Nutrient Requirements

• Most requirements for nutrients that have been published focus on juvenile fish/shrimp

• Many represent single lab experiments, unchallenged, unsupported by others

• Optimum performance can be affected by management, environmental factors and fish/shrimp size

• In formulating diets for a species for which nutrient requirements are unknown, those for a related species are used

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Nutrient Requirements

• Most variation among aquatic species can be associated with whether the animals are: 1) coldwater vs. warmwater; 2) freshwater or marine; 3) finfish vs. crustaceans

• Values in nutrient requirement tables only represent minima, don’t allow for processing or storage losses

• AA’s and minerals are stable, while vitamins and lipids are not stable (affected by heat, oxidation, light, moisture, etc.; store in cool area)

• 50% of ascorbic acid is lost in processing, half-life of 2-3 months in storage

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Physical Properties

• Ground meals are not suitable for feeding to aquatic animals due to poor ingestion, feed conversion, and reduced water quality

• Pellets need to be stable in water until consumed by the target animal• Good pellet stability required for slow-feeding species such as shrimp• Particle size is important to insure appropriate consumption

various sizes of fish feed particles

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Physical Properties

• Diet texture is also important for some aquatic species: some prefer moist vs. dry feeds (e.g., eels vs. salmon)

• sometimes farmers prefer floating pellets, allowing confirmation of ingestion• shrimp prefer sinking pellets (density greater than that of water, 1 g/cm3)• floating feed can be detrimental with respect to consumption by competitors

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Practical Feed Ingredients

• Ingredients used in practical fish/shrimp feeds can be classified as: protein sources (including amino acids) energy sources (COH) lipid sources (also essential fatty acids) vitamin supplements/premixes mineral supplements/premixes growth/pigment enhancers ingredients improving palatability ingredients improving preservation/storage

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Protein SourcesIngredients Crude Protein %

Fish meals > 65 %

Plant proteinsCereals, Pulses, Oil seeds < 25 %Oil seed meals 25 - 48 %

Novel proteinsConcentrates, Isolates, Extracts, Single Cell Proteins, Algae, Insect meals 55 - 70 %

Terrestrial Animal by Products?? > 50 %

Essential Amino Acid Balance Anti-Nutritional Factors

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Fish Meal (FM)

• If made from good quality whole fish, properly processed, it is the highest quality protein source commonly available

• Rich source also of energy and minerals

• Highly digestible, highly palatable, also serves as an attractant

• Usually contains about 65% protein, that is around 80% digestible

• High in LYS, MET, TAU (deficient in plant sources)

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Fish Meal (FM)

• Fish meal also contains 1-2.0% n3 HUFA fatty acids, essential to many fish and all shrimp• If made from by-products, its quality is not as good as trawler-caught

- White fish meal vs. Brown fish meal(whole fish)• Only problem observed: high ash content can sometimes result in mineral imbalance• Used sparingly because of high cost• Can be partially replaced by soybean meal and other animal meals

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Soybean Meal (SBM)

• Soybean meal has one of the best essential amino acid profiles of all protein-rich plant feedstuffs

• SBM does not appear to be deficient in any EAA for catfish• The EAA can be deficient for eels, because their Met/Cys requirement is twice that of

catfish• Some fish find SBM unpalatable, for this reason maximum levels are suggested

- NSP effect

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Proximate composition of various ingredients

1Fish meal LT is fish meal that is dried at a low temperature (Low Temperature), i.e. lower than 70C and with steam. 2Haemoglobin powder consists of red blood cells that are separated from the blood plasma and that are spray dried. 3Wheat is used as a source of starch for the expansion of the pellets (see chapter on extrusion).

http://www.tilapiastichting.nl/Downloads/6%20-%20AHM%20Terpstra%20-%202015%20-%20The%20Composition%20and%20Production%20of%20Fish%20Feeds.pdf

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Percentage of essential amino acids (EAA)1 in fishmeal (FM), rendered meat meal (MM), poultry by-product meal (PBM),blood meal (BM), soybean meal (SBM)

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어분 대체 시 고려 사항

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• Palatability/NSP• Cost• Growth performance/Health• Productivity

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Fish meal alternatives

Ingredients C. protein Price Remarks

Fish meal 68 2300 EAA, EFA, UGF supply

W. gluten 80 2050 Plant protein source

Corn PC (EM75) 78 1700 Plant protein source

Axol-P 76 1500 Hydrolyzed Protein + Feather meal+LCM

Protam (LCM)* 72 1300 Bacterial protein- High lysine

Tankage meal 68 1250 Animal protein/fat source

PBPM* 65 810 Animal protein/fat source

Soy PC 65 2100 Plant protein source- less ANF

Corn gluten 60 720 Plant protein source

FSBM* 55 1300 Plant protein/AA source- fermented

Soybean meal 46 505 Plant protein source

SLP* 45 1100 UGF, PL, Cholesterol

W. flour 13 275 Pelleting binder

Fish oil 1800 EFA, energy supply

Additives Vit, Min, AA, PL, antioxidant, mold inhibitor

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*Poultry by-product meal, Lysine cell mass, fermented soybean meal, squid liver powder.

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Products and by-products from fermentation -bactreria

• BioProtein (Norferm / StatOil)Biomass derived from Methylococcus spp.using methane as a substrate.Protein content was approx. 74% and digestibility on a par with some fish meals. Initially very promising results in salmon with incorporation of 30% of feed.

Norwegian Committee for Food Safety (2006) set a limit of 6% incorporation in salmon feeds

• Protorsan (Ajinomoto)Dehydrated bacterial cells of Cornybacterium glutamicum melassecolaContains about 70% CP and 10% crude fatProduced as a by-product of production of L-glutamic acid by fermentation using sugar-containing substrates e.g. molasses or starch hydrolysates.

• PL73 (Eurolysine)Contains the protein fraction of the bacterium Brevibacterium lactofermentumand is a by-product of L-lysine production

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Soybean Meal (SBM)

• Soybean meal is commonly used to spare fish meal, however, only to a pointtrue for chinook, but not for catfish

• Shrimp will consume high SBM feeds, but diet must be supplemented with fish meal at some point

• Another problem involves losses in energy, minerals and lipids in diets where SBM replaces FM or other animal byproduct proteins

• Another variety of soybean meal is known as “de-hulled”• de-hulled soybean meal contains 25% less ME, 85% less available P and 90% less n-3

FA’s than anchovy meal• soybeans also contain trypsin-inhibitors• trypsin inhibitor reduces digestibility of soy protein by the enzyme trypsin• solution: most soybeans are roasted prior to milling (destroys inhibitor)

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Full-fat Soybeans

• Full fat soybean meal is different from regular SBM in that it has a full fat complement• Fat has not been solvent extracted• 18% fat vs. 0.5%• Often used as an energy source or for general balancing of the formula• Mainly used in salmonid (cold water) fish diets• REM: too high fat = reduced nutrient intake

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Grains and By-products

• Grains are primarily used as COH sources• When whole, they contribute about 62%-72% of dietary starch• Starches are fairly well digested by warm-water species (60-70%), but not by cold• Heating via extrusion improves digestibility by 10=15%• Can also be used as binding agents

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Grains and By-products

• Corn is commonly used in the U.S., but is high in xanthophyll (a pigment), giving tissue a yellow color (not good for fish sales!)

• Corn gluten meal is high in protein (60%) and contains high levels of MET (excellent for formulation)

• Rice bran often used in developing countries due to local rice production• Rice bran is a reasonable COH source, but is high in fiber and fat• Wheat gluten is a good protein source, but too expensive, often used as a

binder

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Animal By-products

• Meat and bone meal is a byproduct of the slaughter house- contains 50-55% crude protein

• Protein quality is low, so only marginally useful and varies dependent upon meat source- can be a good source of energy and P

• Another problem: high ash content• Digestibility improved by flash- or spray-drying• Poultry by-product meal (PBM) is often used by mills also producing

chicken feed• Feather meal high in protein, but indigestible

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Crustacean Meals

• Shrimp waste meal is a reasonably good feed ingredient, if heads are included

• Otherwise, the shell is primarily chitin and of limited digestibility• The ammonia in chitin accounts for about 10-15% of the nitrogen in

whole meal• Also a reasonable source of n-3 fatty acids, cholesterol and

astaxanthin (carotenoid)• Highly palatable and often serves as an attractant in feeds at 1-2%• Others: krill meal, Artemia meal

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Proximate composition of common fish feed materials ranked according to crude protein content (DM)

Sources: Hasan 2009; Skretting; Novus

*Note high levels of “space consuming” ash in seaweeds

Fats and Oils

• Used as energy sources, provide essential fatty acids, attractant, coating of pellet to reduce abrasion

• Both animal and plant fats can be used, animal fats cheaper, better attractants

• Marine lipids often added as oils if FM level is low (otherwise no source of marine FA’s)- sources: menhaden, shark, cod liver

• must be careful in storage of oil, feeds with oils due to rancidification

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Fibrous Feedstuffs

• Most monogastric animals (e.g., fish) do not digest fibrous feedstuffs well• It is unlikely that adding fiber to diets already with more than 3-5% will have

any beneficial effect• High fiber content reduces binding capacity of feeds, inhibits intake (due to

reduced palatability), increases rate of passage and waste production- sources: brans

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Binding Agents

• Binding agents are really needed for pelletized feeds, but not necessarily for extruded feeds - Shrimp pellets by wheat gluten and heat treatments(post-conditioning)

• In extruded feeds, all ingredients are gelatinized by high temperature and bind together well as a result of the process

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Non-nutrient Diet Components

Basic Facts

• In addition to the essential nutrients, feeds may contain organic and inorganic materials that have various effects on aquatic species:beneficial, detrimental or negligible

• They can affect growth, health or the processed product- may be naturally occurring, intentionally or unintentionally added- can be produced via microbial growth

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Toxins and Antimetabolites

• The more important toxins affecting animal feeding are those associated with molds- these are called “mycotoxins”

• Three important genera are Aspergillus, Penicillium and Fusarium• They exist and grow anywhere as long as there is enough COH substrate, no less than

14% moisture, adequate temperature, oxygen• Usually produced in feedstuffs prior to harvest, but also result from poor storage

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Aflatoxin and Ochratoxin

• Aflatoxin is the mycotoxin of greatest concern in feeding of culture species

• both outright toxic and carcinogenic• liver (hepatoma) and blood clotting problems• rainbow trout are highly sensitive at 1 ug/kg

exposure• traditionally, sources include corn, cottonseed

and peanuts• aflatoxin contamination varies year to year

• Ochratoxin are compounds produced by Aspergillus and Penicillium molds

• widely found in nature• typically associated with kidney toxicity• toxic level is 4.7 mg/kg in diet• other mold toxins have been found in warm-

blooded animals, but not in fish• most mold toxins also destroy nutrients in feeds• example: Pseudomonas can separate glutamic

acid from folic acid, making it ineffective

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Microbial Toxins in Commercial Fish/shrimp Feeds

• Usually not known that the feed is contaminated• commercially-processed feeds are less likely to have these toxins• screened against international transport and by feed manufacturers by law• must contain less than 20 ppb• up to manufacturer to require testing• not destroyed by steam pelleting or extrusion• presence in feeds reduced by proprionic acid

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Histamine

• This is a toxic compound found in fish meal, a typical feed ingredient• results from bacterial removal of COOH (carboxylic acid) from the EAA

histidine• comes from improper storage of raw fish prior to production of fish meal• causes a reduction in growth rate• usually comes from “dark” meat portion of fish• other fish meal toxin is “gizzerosine”

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Phytic Acid and Gossypol

• Phytic acid is an organic molecule related to inositol• integral component of plant feedstuffs and holds 60-70% of the phosphorus• problem is, it’s poorly available to fish• reduces availability of zinc, P, Ca etc.

• Gossypol is a component of pigment lands in the cotton plant• limits availability of cottonseed meal used in feeds (suppresses growth rate and

causes liver damage)

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Fish Oils, Fiber

• Marine fish oils contain 20-25% PUFA’s• the “autoxidation” of PUFA’s results in formation of large numbers

of free radicals and peroxide compounds• these are toxic due to reaction with other nutrients, limiting

availability• also cause cellular/subcellular damage• severity of effect reduced by Vit E

• Fiber can also be mildly “toxic” as it increases rate of gut passage• high rate of passage causes reduced availability of nutrients

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Diet Additives:

• Hormonal control used to produce mono sex cultures of fish

- reduces reproduction/increases growth

- ex. Androgenic steroids (ethyltestosterone) fed to tilapia fry

= 90% males

- does not work the same on all fish

• Some feeds can be formulated with antibiotics for treatment of Vibriosis, other bacterial infections

• Three antibiotics approved in U.S. are sulfadimethoxine, sulfamerazine and terrymycin(oxytetracycline, OTC)

• OTC commerically available as “medicated” fish (shrimp) feed, 1,500 mg/kg

• Must not feed medicated diets within 14-21 days from slaughter/harvest (more regulations!)

AntibioticsHormones

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Attractants

• Attractants are materials added to feeds to serve as intake (feeding) stimulants

• They are cost effective since they cause shrimp/fish to eat feeds that otherwise would not be attractive (consumed)

• Facilitates inclusion of by-products

• Usual inclusion level is around 0.5-1.0 %, largely due to cost

• Examples: krill meal, Artemia meal, fish oils, fish meal

• Oxidation of lipids in feeds or feedstuffs can cause reduction of the nutritional value of certain lipids and vitamins

• It can also result in production of toxic free radicals and peroxides

• Potential for formation of these toxic compounds reduced by synthetic compounds such as BHA (butylated hydroxyanisole, BHT (butylated hydroxytoluene)

• Also via natural compounds (Vit E)

Antioxidants

Diet Additives:

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Aquafeed Formulation

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Digestible Energy Digestible Protein

GrowthFeed Conversion

HealthFatty Acids Economy

Environment Amino Acids

Vitamins & Minerals Antinutritional FactorsUnwanted Substances

Formulation

Main Factors Affecting Feed Performance?

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Feed Formulation

• Principle• Cover fish nutritional needs in terms of:

• Digestible energy• DP / DE ratio • Amino acid balance• Fatty acids• Vitamins and minerals

• With a choice of available raw materials

• In a pellet which is:• Dry (<10% H2O, legal limit 14%)• Highly palatable• Strong• Floating or sinking according to needs• As cheap as possible

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Feed Formulation

1 The Linear Equation2 The LP Matrix

LP-based feed formulation software

• has at least three databases: one each for nutrients, ingredients, and products.- Nutrients play the central role in the formulation system because they link both

ingredients and products. The database may be a simple list of nutrients with corresponding units or have additional information such as the species to which the nutrients may be relevant.

- Ingredients: A database on ingredients stores four vital pieces of information: ingredient names, availability of the ingredients for formulation, nutrient composition, and cost.

- Product specifications: Product specifications define the nutrient levels desired in the formula and the ingredient inclusion levels. Either a lower limit and/or an upper limit for each nutrient and ingredient are set

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Nutrients, ingredients, and product specs

Nutrient Unit

Moisture %

Crude protein %

Crude fat %

Crude fiber %

Ash %

Macro minerals (calcium, phosphorus, sodium, etc. Phosphorus is usually expressed on an available basis)

%

Trace minerals (zinc, manganese, iron, copper, selenium, etc.)

mg/kg

Amino acids (arginine, histidine, lysine, methionine, etc. The amino acids are usually expressed on a digestible basis)

%

Fatty acids and other lipid components (linoleic acid, linolenic acid, EPA, DHA, cholesterol, phospholipids, etc.)

%

Starch and nonstarch polysaccharides

%

Energy (usually expressed as digestible energy for aquafeeds)

kcal/kg or MJ/kg

Vitamins (vitamin A, thiamin, riboflavin, etc.)

IU/kg, mg/kg, or µg/kg

Nutrient Unit Value

Dry matter % 88.00

Crude protein % 46.30

Crude fat % 2.70

Crude fiber % 4.50

Ash % 7.20

Digestible energy kcal/kg 3150.00

Calcium % 0.35

Phosphorus % 0.64

Available phosphorus

% 0.25

Sodium % 0.03

Linoleic % 1.25

Potassium % 0.05

Linolenic % 0.16

Arginine % 2.70

Lysine % 2.72

Methionine % 0.61

Met + Cys % 1.24

Threonine % 1.82

Digestible arginine % 2.34

Digestible lysine % 2.30

Digestible methionine

% 0.52

Digestible Met + Cys

% 1.05

Digestible threonine

% 1.59

Choline mg/kg 2706.00

Matrix value of ingredient (soybean meal)List of nutrients used in aquafeed formulation

Nutrient specifications Minimum MaximumCrude protein, % 28Crude fat, % 5.5Crude fiber, % 4Ash, % 15Digestible energy, kcal/kg 2600Digestible lysine, % 1.43Digestible methionine + cysteine, % 0.9Digestible threonine, % 0.74Linoleic acid, % 0.8Linolenic acid, %Available phosphorus, % 0.5Calcium, % 2.5Sodium, % 0.4Potassium, % 1.0Magnesium, % 0.15Vitamin C, mg/kg 50Choline, mg/kg 500

Ingredient specifications, %

Fishmeal, 62% protein

Soybean meal, 46% protein 50Rapeseed meal 10Poultry by-product meal, 55% protein 25Blood meal, spray-dried 5Maize 20 30Rice bran 15Salt 1Limestone 1

Fish oil 0.5

Vitamin premix 0.1

Lysine HCl

dl-methionine

l-threonine

Trace mineral premix 0.5

Mold inhibitor 0.2

Limit for each nutrient and ingredient

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Overview of formulation process

Availability

Price

Nutrient composition

Ingredients Productspecifications

Formulate

Formula results

Production/QA

Analyze and reformulate if

necessary

Nutrient limits

Ingredient limits

Suresh, Aquafeed Formulation (2016)

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Target fish’s nutrient requirements vary:

- Freshwater Vs. Sea water- Sex/Growing stage- Culture condition

TemperatureStocking densityRearing methodWater exchange rate

- Experimental diet composition (purified/practical)

• Expressed still as % diet, not g/kg bw/d

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Formulation of nutrient requirements-based diets are Still a Dream!

Practical diet formulation (EP)

Ingredient 9% FM 6% FM 3% FM 0% FMSBM 32.48 34.82 37.17 39.52FM- Menhaden 9.00 6.00 3.00 -Poultry by-product M 16.00 16.00 16.00 16.00Milo 35.47 33.82 32.33 30.68Corn gluten - 1.67 3.17 4.84Fish oil 3.96 4.22 4.47 4.72DCP 1.50 1.88 2.27 2.65Bentonite 1.00 1.00 1.00 1.00Mold inhibitor 0.15 0.15 0.15 0.15Vitamin premix 0.34 0.34 0.34 0.34Mineral premix 0.08 0.08 0.08 0.08Stay-C 35% 0.02 0.02 0.02 0.0220191126 57

Chemical composition (% fed basis)

Component 9%FM 6%FM 3%FM 0%FMC. protein 35.7 35.9 36.2 36.6C. fat 8.4 8.3 8.6 8.4C. fiber 2.4 1.8 2.1 1.9C. ash 8.2 7.9 7.9 8.1Ca 1.3 1.3 1.2 1.2P 1.2 1.2 1.2 1.3Lys 2.0 2.0 1.9 1.8Met+Cys 1.1 1.1 1.1 1.1%protein from plant sources 54.9 60.4 66.0 71.5%protein from animal sourecs 45.1 39.6 34.0 28.5

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Trophic-level value of agriculture, aquaculture and fishery(Durate et al., 2009)

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Marine Fish Farming

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Fermentation

SCP, Lysine & Cell mass

Reduction of Trophic-level value (Microbial protein to fish)

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Reduction of Trophic-level value (Algal fatty acids to fish)

EVONIK, 2017

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TERIMA KASIH

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