UNIVERSITI PUTRA MALAYSIA
NUTRITIVE VALUE AND BIOAVAILABILITY OF PROTEIN FEEDSTUFFS FOR BROILER CHICKENS
FAIRDA LIM
FPV 1985 2
This is to certify that we have read this thesis
entitled "Nutritive Value and Bioavailability of Protein
Feedstuffs for Broiler Chickens" by Fairda Lim, and in
our opinion it is satisfactory in terms of scope, quality
and presentation as partial fulfilment of the requirements
for the degree of Master of Science.
Assoc. Pr
ZAINUDDIN, Ph.D.
ssor/Dean of Graduate Studies
Universiti Pertanian Malaysia
(Chairman Board of Examiners)
0?LP�( I.R. SIBBALD, D.Sc.
Chairman, Poultry Nutrition Program
Animal Research Centre
Canada
(External Examiner)
Professor/Deputy Vice-Chancellor
Universiti Pertanian Malaysia
(Internal Examiner) j)
----I -�" ." rt!u'/';::>hd� P. V I JCHULATA, Ph.D.
Lecturer
Faculty of Veterinary Medicine & Animal Sciences
Universiti Pertanian Malaysia
(Internal Examiner)
Department of Science
Faculty of Veterinary Medicine & Animal Sciences
Universiti Pertanian Malaysia
(Supervisor)
This thesis was submitted to the Senate of Universiti Pertanian Malaysia and was accepted as partial fulfilment of the requirements for the degree of Master of Science.
ALA��DDIN' Ph.D. Associate Professor/ Dean of Graduate Studies
NUTRITIVE VALUE AND BIOAVAILABILITY OF PROTEIN FEEDSTUFFS FOR BROILER CHICKENS
THESIS
A thesis submitted in partial fulfilbnent of the requirements for the Degree of Master of Science
at Universiti Pertanian Malaysia
by
FAIRDA LIM
Faculty of Veterinary Medicine and Animal Science Universiti Pertanian Malaysia
Serdang, Selangor, Malaysia 1985
ACKNOWLEDGEMENT
The author expresses her sincere gratitude and appreciation
to her supervisor , Professor Dr. Rudy I . Hutagalung , for his
valuable supervision and guidance in the planning and implementation
of experiments and the preparation of dissertation , and Dr . Yeong
Shue Woh , as a consultant and the provision of facilities during
the course of experimental work .
The author is also grateful to the Direc tor-General of MARDI
for providing scholarship and study leave to undertake the higher
degree programme at the Universiti Pertanian Malaysia .
Sincere thanks are extended to Dr . V . Rhagavan of the Federal
Flour Mill for the supply of FFM soybean meal and Mr . Sunny Heng
Kee Siang , the Manager for Export and Development Division ,
Industrial Business Area , I . C . I . (Malaysia ) Sdn . Bhd . for the
supply of "Pruteen" for the experiments .
Special gratitude and appreciation is due to Mr . Ong Hwee Keng
for his motivation throughout the programme.
Special thanks are also extended to the Staff of MARD I,
including all personnel in the Animal Nutrition Laboratory ,
Mr. Ahmad Shukri bin Haji Othman for the analysis of data and
Mr . Ong Boon Thuan for his amino acid analyses .
Last but not least , apprec iation is also due to Dr . C . Devendra
for his encouragement.
ii
TABLE OF CONTENTS
TITLE
ACKNOWLEDGEMENT
TABLE OF CONTENTS
LIST OF TABLES
LIST OF FIGURES
LIST OF APPENDIX TABLES
LIST OF ABBREVIATIONS
ABSTRACT
CHAPTER I INTRODUCTION
CHAPTER II LITERATURE REVImoJ
MET HODS OF FEED EVALUATION
I. Ch emical evaluation
(i) Proximate analysis
(ii) Gross energy
(iii) Minerals
(iv) Fibre analysis
(v) Amino acid
(vi) In vitro evaluation
2. Feeding values of proteinaceous feeds tuffs
Page
�
vii
viii
x
3
3
3
6
6
8
8
9
Comparative nutritive values of some 10 protein sources
Fish meal 10
Soybean meal 11
Pruteen 12
iii
3. Biological evaluation
(i) Bioavailable energy and determination
Page
13
13
Apparent Metabolizable Energy 14 (AME)
True Metabolizable Energy (TME) 15
Indirect measurement of ME 16 Selection of a bioassay 16
(ii) Amino acid availability (AAA) and determination
Biological assays
Enzymatic assays
Chemical assays
Microbiological assays
Faecal and ileal assays
AAA from TME determination
17
1 9
21
21
22
23
24
CHAPTER III EXPERIMENT I
Introduction 25
Materials and Methods 25
Results 26
Discussion 32
CHAPTER IV EXPERIMENT 2
Introduction 3 5
Experiment 2a 35
Materials and methods 3 5
Results 40
Discussion 47
Experiment 2b 52
iv
CHAPTER V
Materi al s and method s
Re sul ts
D i scus s ion
EXPERIMENT 3
Introduc tion
Materials and method s
Results
Discuss ion
CHAPTER VI GENERAL D ISCU S S ION
CHAPTER VII SUMMARY AND CONCLU SION
BIBLIOGRAPHY
APPEND ICES Appendix
Appendix Tab les
v
Page
52
55
6 1
64
64
6 7
7 1
7 6
7 8
7 9
Al
A4
TABLE
2
3
4
5
6 .
7
8
9
1 0
1 1
1 2
1 3
1 4
LIST OF TABLES
Chemi cal comp osi tion of pr otein sour ces
Amino acid profi les of protein sources
In vitro digestibility of proteinanceous feed stuf f s
Composition of starter diet s in Experiment 2a
Composition of f inisher diet s in Experiment 2a
Effects of fish meals and Pruteen on the performance of broi ler chickens (8 weeks)
Eff ect s of dietary levels of f i sh mea l s and Pruteen on apparent digestibilities of the starter diet s by br oiler chickens
Compo sition of starter diet s in Experiment 2b
Comp osit ion of f ini sher diet s in Experiment 2b
Eff ects of level and type of soybean mea l s on the performance of broi ler chickens (8 weeks)
Effects of dietary level and typ e of soybean meals on apprent digestibi lities of the starter diet s by broiler chickens
True metabolizable energy (TME ) and nitrogencorrected met aboliz ab le energy TMEn) values
Amino acid avai lability (% DM) of proteinanceous f eedstuff s in chi ckens
Amino acid and true avai lable amino acid (TAAA) of proteinaceous feedstuf f s
vi
PAGE
27
30
3 1
3 7
3 8
44
45
53
54
5 9
6 0
6 8
6 9
7 0
LIST OF F IGURES
FIGURE PAGE
1 Average body weight through 8 weeks , Experiment 2a 41
2 Feed intake through 8 weeks , Experiment 2a 42
3 Feed /gain through 8 weeks , Experiment 2a 43
4 Average body weight through 8 weeks , Experiment 2b 56
5 Feed intake through 8 weeks , Experiment 2b 5 7
6 Feed/gain through 8 weeks , Experiment 2b 5 8
vii
TABLE
1
2
3
4
5
6a
L IST OF APPENDIX TABLES
Chemical analyses of starter diets , Experiment 2a
Chemical analyses of finisher diets , Experiment 2a
Chemical analyses of starter diets , Experiment 2b
Chemical analyses of finisher diets , Experiment 2b
Data on digestibility trial , Experiment 2a
Data on total weight gain , performance trial of Experiment 2a
6b Data on total intake , performance trial of Experiment 2a
PAGE
A 4
A 4
A 5
A 5
A 6
A 7
A 8
6c Data on feed/gain , performance trial of Experiment 2a A 9
7 Data on digestibility trial , Experiment 2b AIO
8a Data on total weight gain , performance trial of Experiment 2b All
8b Data on total intake , performance trial of Experiment 2b A12
8c Data on feed/gain , performance trial of Experiment 2b A13
9 Feed and faecal data for true metabolizable energy determination
10
11
12
13
14
15
Amino acid composition of feed and faecal samples (% DM )
Analysis of variance on digestibility of DM , Experiment 2a
Analysis of variance on digestibility of C P , Experiment 2a
Analysis of variance on digestibility of EE , Experiment 2a
Analysis of variance on digestibility of CF, Experiment 2 a
Analysis o f varianqe on digestibility of NFE , Experiment 2a
viii
A14
A15
A16
A16
A17
A17
A18
TABLE PAGE
16 Analysis of variance on total weight gain , Experiment 2a A18
17 Analysis of variance on total intake , Experiment 2a A19
18 Analysis of variance on feed/gain , Experiment 2a A19
19 Analysis of variance on digestibility of DM, Experiment 2b A20
20 Analysis of variance on digestibility of CP , Experiment 2b A20
21 Analysis of variance on digestibility of EE , Experiment 2b A21
22 Analysis of variance on digestibility of CF , Experiment 2b A21
23 Analysis of variance on digestibility of NFE , Experiment 2b A22
24 Analysis of variance on total weight gain , Experiment 2b A22
25 Analysis of variance on total intake , Experiment 2b A23
26 Analysis of variance on feed/gain , Experiment 2b A23
ix
AAA AAAA ADF ADL AME BE CF CP DCP DM EE ( FE + UE ) ( FEn + UEn ) FFM ( FN + UN ) FSM (FmE + UeE ) ( FmEn + UeEn ) g GE IE IN kcal kg ME MJ N NDF NE NFE SBM SS TAAA TME TMEn
LIST OF ABBREVIATIONS
amino acid availability apparent amino acid availability acid detergent fibre acid detergent lignin apparent metabolizable energy bioavailable energy crude fibre c rude protein d igestible crude protein dry matter ether extract excreta energy excreta energy corrected to zero nitrogen balance Federal Flour Mill faecal and urinary nitrogen fish meal Metabolic faecal plus endogenous urinary energy nitrogen corrected energy excretion of unfed bird gram gross energy energy input nitrogen input kilocalorie kilogram metabolizable energy megajoule nitrogen neutral detergent fibre net energy nitrogen free extract soybean meal Soon Soon true amino acid availability true metabolizable energy TME corrected to zero nitrogen balanc e
x
ABSTRACT
An abs tract of the thesis presented to the Senate of
Universiti Pertanian Malaysia in partial fulfilment of the
requirement for the Degree of Mas ter of Science .
NUTRITIVE VALUE AND BIOAVAILABILITY OF PROTEIN FEEDSTUFFS
FOR BROILER CHICKENS
by
FAIRDA LIM
Augus t , 1 985
Supervisor: Professor Dr . Rudy I . Hutagalung
Faculty: Kedoktoran Veterinar dan Sains Peternakan
Three experiments were conducted to evaluate the nutritive value
and biological availability of nutrients in proteinaceous feedstuffs
for broiler chickens. These include Thai fish meal ( FSM ) , Peruvian
FSM, China soybean meal ( SBM ) , two locally-processed SBM , Federal
Flour Mill ( FFM ) and Soon Soon ( SS ) , respectively, and single-cell
protein "Pruteen" .
In Experiment 1 , the nutritive values of these feedstuffs were
subjected to the following chemical analyses: proximate, minerals,
detergent fibre, amino acids and in vitro digestibility . C rude
protein ( C P ) contents of fish meals ( on dry matter basis ) were
xi
54 . 1 7% for Thai FSM and 60. 98% f or Peruvian FSM. Ash cont ent in
Thai FSM was higher ( 25.86%) than Peruvian F SM (15 . 5 1 %) . Gros s energy
in Peruvian FSM ( 4 , 488 kcal/ g ) was higher than Thai FSM (3 , 78 1 kcal/g ) ,
attributing mainly to higher fat content in the Peruvian FSM ( 1 0 . 3 7%)
than in the Thai FSM ( 7 . 5 9%) . In vitro evaluation of fish meals
showed that the digestible CP (DCP ) was higher for Thai FSM ( 4 2 . 2%)
than the Peruvian FS�1 (40 . 0%) . SBM of China origin showed highest
protein content (44 . 4 1 %) f ollowed by FFM SBM (43 . 09%) and SS SBM
( 4 1 . 04%) . However , the amino acid profile s showed that China SBM
had the lowest total amino acid value ( 42 . 8 1 % ) and SS the highest
(44 . 08%) . In vitro evaluation showed that China SBM had the
highest DCP (38 . 33%) and that of the two locally-proces sed SBM were
comparable (35 . 1 5 and 35 . 64%) . Pruteen had a high CP value ( 7 1 . 94%)
and DCP (6 1 . 0%) , but low in crude fibre (! 1 %) , and high in gro ss
energy (4 , 5 2 7 kcal/g ) .
Experiment 2 consisted of two trial s . In the first trial , the
effects of Thai FSM (5 to 3 2 % ) and Pruteen ( 3 and 5%) inc lusion �n
the diets on digestibility and performance by broiler chickens were
studied . The results showed that diets cont aining FSM exhibited higher
nutrient digestibility and superior performance than the contro l diet
without added FSM . Optimal performance was obtained at 1 0 and 1 5% FSM
inclusion . In the second trial the inclusion of Peruvian FSM ( 28/22%)
was inferior to Thai FSM ( 3 2 / 25 % ) in nutrient digestibility and
p erf ormance . Pruteen gave comparable nutrient digestibility and
p erformance as tho s e on FSM-based die ts , although it was observed that
there was growth depression and low f eed intake at higher levels of Pruteen
inc lusion. The second trial s tudied the effect of type and level
xii
of SBM-based diets on the digestibility an d performance by broiler
chickens. The data showed that digestibility and performance were
superior at lower level (20%) of SBM inclusion in diets containing
FSM than higher level (44/35%) of SBM with no added FSM. This
study revealed that SS SBM had relatively lowest quality among the
three SBM.
Experiment 3 was carried out to determine the true metabolizable
energy (TME) as bi oavailable energy� and amino acid availability of
protein feedstuffs. The results showed that Thai FSM contained
3.30 kcal TME/g and Pruteen , 3.23 kcal TME/g. The TME for FFM SBM
was the highest (2.92 kcal!g) and China, the lowest (2.69 kcal/g)
of the SBM. Thai FSM and Pruteen showed high true amino acid
availability (TAAA) (average 89.0 and 92.0%, respectively). The
TAAA of China SBM was lowest (74.1%) whereas the two locally-processed
SBM had the same TAAA (86.7 and 86.4%, respectively).
xiii
CHAPTER I
INTRODUCTION
Efficient animal production depend s on the availability of
feedstuf f s of good qua lity and undoubt ly protein qua lity is an important
measure . The quality of protein source is the mo st variable among
feed ingredient s and also g eneral ly , the more expensive in cost .
Therefore , the study of feed protein quality deserves thorough
assessment , invo lving the study of the general quality of feed stuffs ,
the amino acid composition and the bioavailabi lity of nutrient s to
the animals .
Chemical analyse s provide e stimates of the nutritive quality of
feed stuff s and p lay an important part in providing information on the
amino acid profiles of feed stuf f s . Bioassay , digestibility and
performance trials need to be included to assess the nutritive value
of feed stuffs adequately. Bioavailable nutrients such as amino acids
and energy in feedstuff s are of fundamental importance in determining
value . Knowledge of the amount and availability of various nutrient s
supplied by the feedstuff s is essential in order to avoid inefficient
feeding and enormous was tage of feed re source s .
The determination amino acid availability is time-consuming and
expensive . This has delayed the publication of data on available
amino acid requirements and the formulation of diet s based on available
am�no acids . It i s desirable t o emp loy a simp ler and more rapid
method in determining the amino acid availabi lities in f eedstuf f s .
True metabolizable energy (TME), an estimat e of bioavailable
energy , is important in estimating requirements and formulating rations .
A rapid bios s ary f or TME in feedstuff s for poultry has been
developed (Sibbald , 1 97 6 ) and an adaption of the TME procedure
(Likuski and Dorrell , 1 97 8 ; Sibbald 1 97 9b , c ) can be used to assay
for amino acid availability (AAA) .
Optimal bird performance depend s very much on diet formulation
of correct amount of the various nutrient to meet accurately the
quantitative nutritional requirement s . Emphasis is placed on the
relationships between compositional attribut es and animal performance .
Thus , the nutrient content and availability in feed s tuf f s is
becoming important in livestock feeding and needs to be considered
for precise diet formulation and preparation .
Feed f ormulation in Malaysia , at present relies on chemical
compo sition and overseas nutrient availability data. It is not real ly
appropiate to use the se value , in view of the variabi lity due to
variety, location and methods of produc tion of the locally available
proteinaceous feedstuf f s . However , work on nutrient availability
of the locally available proteinaceous feedstuffs is still very much
limited in Malaysia . Consequent ly, thi s study seeks to elucidate
by chemical and biological evaluation , the quality, the TAAA and the
TME of some of the locally availabl e proteinaceous feedstuff s used
Ln broiler diet s . The se inc lude various fish meal s and soybean meals .
In addition , the recent ly introduced single ce l l pro tein in Malaysia ,
designated as Pruteen, produced from Methylophi lus methylotrophus
by anaerobic fermentation of methanol in the presence of ammonia was
chosen for evaluation .
2
CHAPTER II
LITERATURE REVIEW
A. METHODS OF FEED EVALUATION
Evaluation of feedstuff s �n the context of animal
production is very important for effici ent feeding to improve
performance of the animals and to avoid enormous wastage of
feed resources . The simp lest evaluation of feed stuff s is a
compari son of their gross ef fects as in feeding trial s . A
further ref inement i s the chemical analyses of feed stuf f s and
subsequently, determination of the digestible nutrient s and
bioavailabi lities in the different feed stuf f s . The se chemical
and bio logical means of evaluating feeds tuf f s are accepted
standardised methods of provid ing inf ormation on composi tion
and/or nutritive values .
1 . Chemical Evaluation
Chemi stry is very important in predicting the value
of feeds tuf f s . Chemical analyse s provide means to det ermine
the common nutrient s such as protein and its amino acid
profi le , f ibre , fat , ash, minera l s and ca lor if ic content .
These procedures are described in the book Off icial
Methods of Ana lyses , publi shed by the Association of Official
Analytical Chemi s t s (AOAC , 1 9 75 ) .
(i) Proximate Analysi s
The need t o deve lop and quantitative system
3
f or classifying and evaluating feedstuf f s has
long been recognized by anima l nutritionists .
Henneberg and Stohman , Horking in the ear ly par t
o f the nineteenth century at the Heende
Experimental Station in Germany , introduced the
'Heende' Sys tem of feed analysis which de scribes
feeds in terms of their proximate components :
moisture , ash, fat , fibre , crude protein and
nitrogen-free extrac t . At present , the
proximate analysi s is probab ly the mo st
generally used chemical scheme for de scribing
feed stuff s .
The Heende analysis does not define the
nutrient content of feed s tuff s . It is an index of
nutritive value only because the fractions that it
isolates are correlated with some of the properties
of feeds that have nutritional significance .
Consequent ly, it is a useful de scriptive device in
establishing the characteristics of feedstuf f s .
Al though the Heende analysis is inadequate
as a measure of the chemical composition, it provides
useful information on the nutritive value of feedstuf f s
in general and i s s ti l l re tained a s a routine method
of analysis . In addition , mos t laboratories
supp lement the data obtained from proximate
analysis with more detai led analyses carried out by
modern technique s .
4
The general princip les of pr oximate analysis
are given below : -
(a)
(b)
(c )
(d )
(e)
Crude Pro te in (CP) This is approximate by
mUltip lying the Kj eldahl ni trogen value by the
factor 6 . 25 . Dep ending on the type of
feedstuff , a greater or les ser part of the
ni trogen is pre sent as true protein while
the remainder i s inorgani c ni trogen salts ,
amides , etc .
E ther Extract (Crude Fat ) (EE ) This i s
measur ed as diethylether or petro leum ether
extracted material . It includes no t only oils
and fats but fatty acid s , resins , chlorophyl l ,
e t c .
Crude Fibre (CF ) Thi s refers to organic
mat ter inso lub le Ln hot di lute sulphuric acid
and dilute sodium hydr oxide solution . It is
noted that the CF in many cases is a mis leading
indication of the diges tibility and nutritive
value of a feedstuff as has been shown by
the development of methods for partitioning
the f ibre frac tion (Van Soest , 1 96 7 ) .
Ash This i s de termined by igni ting samp les
unti l they are free of carbon .
Nitrogen Free Extract (NFE ) This i s the
difference between the sum of the other
5
cons ti tuents and the original dry weight .
In other words , it is what remains after the
other group s of comp onents have been detected
by analysis. NFE inc ludes sugar s , s tarche s,
etc .
(ii) Gross Energy
The animal obtains energy from its feed s . The
energy present in the feed i s measured by converting
i t into heat energy and determining the heat produced .
The conver sion is carried out by oxidizing the feed
by oxidizing the feed by burning i t ; the quanti ty of
heat resulting from the comp lete oxidation of unit
weight of a feedstuff � s known as the gross energy
or heat of combus tion of that feedstuff .
(iii) Minerals
It i s important to know mineral contents of
feedstuff s because of the great inherent variabi lity .
Macro-elements like calcium (Ca) , pho sphorus (P) and
magnesium (Mg ) may be measured . The ashed samp les
are digested in acids and analysed for the
miner al s . C a and Mg are determined by atomic
absorption, and P i s determined by co lor imetric
method , involving the vanadomolybdate comp lex .
(iv) Fibre Analysi s
Crude f ibre has been and remains a common
means of evaluating f ibre content in feedstuff s .
6
However , the need f or bett er methods of the fibre
analysis arises due to the def i ci encies of crude
f ibre analysi s and asso ciat ed proximate analysis
(Van Soe st , 1 977 ) . The we aknes ses of the CF method
have been known for a long time but have on ly
re cent ly be come widely re cognized . The err or of the
CF assay stems from it s f ai lure to re cover lignin
and other f ibre comp onent s (Van Soest , 1 97 5 ) .
Fibre s are variable in their comp os it ion
and properties , and it is not p os sible t o des cribe
the characterist i cs and' amou�t of fibre with a
sing le value. They wi l l require ana lysis f or
comp onent s in cluding lignin , ce l lulose and hemi ce llu lose .
The Deterg ent System of Fibre Analyses
The obj ect ive of the detergent system of ana lys es
is to fr acti onate f eeds of p lant origin re lat ive to
their nutritive avai labi lity and f ibre content .
f The ce ll wa ll comp onent s of feed ar e re covered l.n the
neutral detergent f ibre (NDF ) , whi le the acid
detergent f ibre (ADF ) divides the se int o fract ions 1.1
s o lub le and inso lub le in IN acid. The acid s o luble
fract ion include s the hemi ce l lu lose and ce l l wal l prote in
whi le the residue (ADF ) re cover s ce l lu lose and lignin
whi ch is the least digestible non-carbohydrate
fractions . Improved systems f or f ibre ana lysi s
(Goering and Van Soe st , 1 970) have been wide ly
accepted in resear ch laborat ori es as r ep la cement s
7
f or the older crude f ibre trechnique .
Studies have been conduc ted in the use of the
detergent f ibr e analyses in formulating broiler
rations (Strong and Ho lder , 1 97 8 ) . The study
concluded that ADF is a more accurate and
repeatab le analys i s for fibre than 1S CF . Thus ,
when data are avai lable , ADF might be preferable to CF
in formulating poultry diet s .
(v) Amino Acid
The crude protein value doe s no t distinguish
between protein and non-protein ni trogen and as sumes
that all protein contains 1 6% ni trogen and i s of
equal bio logical availability . I t gives no
indi cation of the content of amino acid s which constitute
the protein and which are required by the animal . The
amino acid composi tion and levels are useful in
predicting the potential nutri tive value of the
pro tein to the animal .
The amino acid composi tion and levels can
be determined accurately by ion-exchange
chromatography of protein hydrolysates .
(iv) In Vitro Evaluation
� vitro evaluation provides a means of rap id
assessment of nutrient bioavai lability . Procedures
have been developed and modified to imitate the
action of the animal digestive system and are used
8