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