ONLY FOR EDUCATIONAL PURPOSES.

1. Evaluation of
Aquafeed ingredients
and Feed Quality
Sourav Deb Sharma
M.F.Sc Scholar
ICAR-CIFE

2. Table of contents
01
02
04
03
Introduction
Biological evaluation
Physical evaluation
Chemical evaluation
05
Conclusion

3. INTRODUCTION
 In a recent time fish has become
important source of high quality protein
at low cost for human consumption and
also plays role in the prevention and
treatment of various diseases.
 Horizontal expansion of the aquaculture
is limited due to non availability of land
for fish culture.
 One of the input for increasing the
production is using high quality feed.
 Using the commercial feed in aquaculture
increase the cost by 60 percent of total
fish production cost.

4. PHYSICAL
EVALUATION
01

5. PHYSICAL EVALUATION
 Smell, taste, sight, touch.  The compound microscope: To identify
the internal structure of feed components
 The stereomicroscope: To identify the
external structure.
Techniques for sample
preparation
 Screening
 Floatation
 Measurement of how fast and how
much of the feed will be taken by the
aquatic animal.
 Explore opportunities for growth and expansion of
the product or service
Use of senses Feed Microscopy
Bulk density of feedstaffs
 Must be dry, free flowing, and
uniform in appearance.
Attractability

6. Physical evaluation is easy but
rough in nature. One must be
highly trained to identify the
changes in the nature of the raw
material/feeds.
01
Adulteration:- Admixture of a pure
substance with some cheaper and
low quality substance. It is done
intentionally usually to make money.
02
CONT.

7. CHEMICAL
EVALUATION
02

8. Moisture
CHEMICAL EVALUATION
Feed
sample
Moisture
free sample
Ash
Crude
protein
Fat free
Residue
Crude
fibre
Crude fat
Oven dry at 105º C
Ether extraction
Incineration at 550º C
Digestion
Distillation
Titration
Boil in weak acid
Boil in weak alkali
Oven dry at 105º C
Incineration at 550º C
Proximate analysis

9. ALPINE SKI HOUSE
BIURET METHOD
• Spectrophotometric method.
• Applicable for extracted liquid fish protein aliquotes.
• Biuret test is used to detect protein as it can detect peptide
bonds between amino acids.
9
lowry METHOD
 This method estimates total protein of feedstuffs
and feeds.
 It uses Folin-Ciocalteu reagent and has a blue end
color with extinction at 660 nm.
 The principle behind this method is the reduction
of the phosphomolybdic acid-phosphotungstic acid
(Folin-Ciocalteu) reagent by tyrosine and
tryptophan residues in the protein.
METHOD OF PROTEIN EVALUATION
TRUE PROTEIN VALUE DETERMINATION

10. • Amino acid analysis is done using either an amino acid analyzer (AAA) or high
performance liquid chromatograph (HPLC) with fluorescence detector.
• HPLC utilizes o-phthalaldehyde/N-acetylcystine (OPA/AcCys) and Na
hypochlorite reagent.
• The amino acid is identified in the resulting chromatogram by the retention time
of the peak and quantified by the area under such peak.
DETERMINATION OF PROTEIN QUALITY
In the amino acid analysis by AAA,
ninhydrin solution is used for color
development instead of OPA/AcCys.
CONT.
Amino acid composition

11. ALPINE SKI HOUSE
• The EAAI method of chemically evaluating a protein
considers all essential amino acids rather than the most
limiting amino acid with respect to some standard.
• EAAI is the geometric mean of the amino acid found by
comparing the content of the ten essential amino acids in a
feed protein with that found in fish tissue protein.
11
Essential Amino Acid Index(EAAI)
CONT.
In which a, b, ... j are the percent of essential amino acids in the
food protein and ae, be,… je are the percent of the respective
amino acids in whole egg protein.

12. ALPINE SKI HOUSE
• The quality of lipid contained in feedstuffs and feeds
is determined by its fatty acid composition which is
obtained through gas chromatographic (GC) analysis.
• It is a technique of separating sample into its
constituents and then measuring or identifying
components.
• This technique requires that samples should be
volatile.
12
FIG: A sample gas chromatographic analysis
showing retention times of various fatty acids
in a feed sample.
METHODS OF LIPID EVALUATION
MEASUREMENT OF LIPID QUALITY

13. 13
13
TESTS FOR LIPID RANCIDITY
• Peroxide value (PV): Peroxide value is
defined as the reactive oxygen content
expressed in terms of milliequivalents
(meq) of free iodine per kg of fat..
01
• Free fatty acid values may be determined
using the titrimetric method. The presence of
these free fatty acids can speed up oxidative
deterioration.
02
• Thiobarbituric acid number (TBA): This
method involves heating the oil with
thiobarbituric acid in the presence of a
strong acid.
• A red coloration is produced as a result of a
reaction between the thiobarbituric acid
and malonaldehyde.
03

14. METHOD OF VITAMIN EVALUATION
• High Performance Liquid Chromatography (HPLC).
The HPLC is a powerful analytical method or tool
with a wide range of applications.
• It provides high resolution, sensitivity, and automatic
operation, and is efficient in determining the amount
of vitamins contained in feedstuffs and feeds
FIG: A sample HPLC chromatogram of a vitamin
mixture

15. 15
15
METHODS OF ENERGY DETERMINATION
• Digestible energy(DE):.
01
• Metabolizable energy(ME): The ME is the
portion of the gross energy consumed by the
fish for growth, fattening, or heat
production.
• It does not appear in the feces or in the urine
of fish.
02
• Net Energy(NE): This represents the most
precise measure of the energy needs of fish
and the capacity of different feeds to meet
these needs.
• NE values allow for the energy lost in the
urine and feces, but they also take into
consideration the energy lost as heat during
nutrient utilization.
03
The caloric content of the feed sample is determined
through the use of a bomb calorimeter..

16. ALPINE SKI HOUSE
16
Urease activity
ANALYSIS OF TOXINS IN FEEDS
• Found in mainly raw soybean.
• Heat treatment of soybean meal at 120º C to
remove this enzyme.
• The urease activity of soybean mat be determined
by pH increment method, titration method and
Nash reagent method
Aflatoxin
• Class of potent toxins produced by the mold
Aspergillus flavus .
• Sources: Groundnut cake, copra, peanuts, corn, rice,
and legumes produced and stored under hot and
humid conditions.
• The chromatographic methods most widely and
routinely used for aflatoxin analysis.
Gossypol
• Endogenous toxin present in the gland of
cottonseed which persists during meal production
unless removed by a special process, or, unless the
cottonseed is a glandless variety.
• Sources: Cotton seed meal
 glandless (gossypol level - 0.01%)
 glanded variety (avg, 1.3%)
• The gossypol level in a feed ingredient is
determined spectrophotometrically.
Permissible limit: <0.01%

17. BIOLOGICAL
EVALUATION
03

18. 18
18
PARAMETERS TO BE MONITORED IN FEEDING
Absolute growth, wt gain = wf - wi,
where: w f = final wt; wi = initial wt
Absolute growth rate=
𝑤𝑡 𝑔𝑎𝑖𝑛
𝑑𝑎𝑦
=
𝑤𝑓−𝑤𝑖
𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑤𝑡
.
01
Relative growth
% 𝒘𝒕 𝒈𝒂𝒊𝒏 = 𝒘𝒇 − 𝒘𝒊 ∗
𝟏𝟎𝟎
𝒘𝒊
02
Relative growth rate
% 𝑤𝑡 𝑔𝑎𝑖𝑛
𝑑𝑎𝑦
= 𝑤𝑓 − 𝑤𝑖 ∗
100
𝑤𝑖 𝑑𝑎𝑦𝑠 𝑜𝑓 𝑐𝑢𝑙𝑡𝑢𝑟𝑒
03
Specific growth rate(SGR)
𝑺𝑮𝑹 = (𝒍𝒏 𝒘𝟐 − 𝒍𝒏 𝒘𝟏 )/(𝒕𝟐 − 𝒕𝟏))
04
Growth

19. 19
19
CONT.
Feed conversion ratio(FCR)
𝑭𝑪𝑹
= ((𝑀𝑎𝑠𝑠 𝑜𝑓 𝑓𝑜𝑜𝑑 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑 𝑑𝑟𝑦 )
/((𝐼𝑛𝑐𝑟𝑒𝑎𝑠𝑒 𝑖𝑛 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑎𝑛𝑖𝑚𝑎𝑙 𝑤𝑒𝑡 )
.
01
𝑭𝒆𝒆𝒅 𝒆𝒇𝒇𝒊𝒄𝒊𝒆𝒏𝒄𝒚 %
= 𝒘𝒕 𝒈𝒂𝒊𝒏 ∗
𝟏𝟎𝟎
𝒅𝒓𝒚 𝒇𝒆𝒆𝒅 𝒄𝒐𝒏𝒔𝒖𝒎𝒆𝒅
02
% 𝑫𝒊𝒈𝒆𝒔𝒕𝒊𝒃𝒊𝒍𝒊𝒕𝒚
= 𝑵𝒖𝒕𝒓𝒊𝒆𝒏𝒕 𝒂𝒃𝒔𝒐𝒓𝒃𝒆𝒅 ∗
𝟏𝟎𝟎
𝒏𝒖𝒕𝒓𝒊𝒆𝒏𝒕 𝒄𝒐𝒏𝒔𝒖𝒎𝒆𝒅
03
Efficiency of feed utilization
Digestibility of nutrients

20. 20
20
CONT.
Protein efficiency ratio(PER)
𝑃𝐸𝑅 =
𝑤𝑡 𝑔𝑎𝑖𝑛
𝑝𝑟𝑜𝑡𝑒𝑖𝑛 𝑖𝑛𝑡𝑎𝑘𝑒
.
01
𝑩𝒊𝒐𝒍𝒐𝒈𝒊𝒄𝒂𝒍 𝒗𝒂𝒍𝒖𝒆 𝑩𝑽 =
𝑵 𝒓𝒆𝒕𝒂𝒊𝒏𝒆𝒅
𝑵 𝒂𝒃𝒔𝒐𝒓𝒃𝒆𝒅
02
𝑵𝒆𝒕 𝒑𝒓𝒐𝒕𝒆𝒊𝒏 𝒗𝒂𝒍𝒖𝒆 𝑵𝑷𝑽 𝒐𝒓
𝑷𝒓𝒐𝒅𝒖𝒄𝒕𝒊𝒗𝒆 𝒑𝒓𝒐𝒕𝒆𝒊𝒏 𝒗𝒂𝒍𝒖𝒆 𝑷𝑷𝑽
= 𝑩𝑽 ∗ 𝒅𝒊𝒈𝒆𝒔𝒕𝒊𝒃𝒊𝒍𝒊𝒕𝒚
03
Efficiency of protein utilization

21. CONCLUSION
• The choice of high quality feedstuffs for incorporation
into aquafeeds is crucial to the success of an
aquaculture venture.
• Proper methods of feedstuff and feed evaluation
should be learned and applied to attain an effective
feed. The systematic evaluation of feedstuffs and feed
includes physical, chemical, and biological methods.
• Among these methods, the biological method which
involves actual feeding experiments gives a more
accurate estimate of feed utilization.

22. sourav.fnftmb306@cife.edu.in
THANK YOU
SOURAV DEB SHARMA