Indonesian Journal of Nutrition

and Feed Science

Vol. 2 No. 1, January 2011

Editorial Board : Toto Toharmat (Editor in Chief)

A. R. Alimon
Ali Agus
Arnold Sinurat
E. R. Ørskov
Nahrowi
Komang G. Wiryawan
Yantyati Widyastuti

Administration : Ratih Windyaningrum

and Secretary Sri Suharti

Editorial Address : Faculty of Animal Science

Department of Nutrition and Feed Technology
Bogor Agricultural University (IPB)
Campus IPB Dramaga, Bogor 16680, Indonesia
Phone : (0251) 8626419, e-mail : jurnal_aini@yahoo.com

Indonesian Journal of Nutrition and Feed Science, published original papers in the field of nutrition
and feed science. Manuscript submitted for publication and inquires regarding them should be sent to
the Editorial Board, Indonesian Association of Nutrition and Feed Science (AINI), Faculty of Animal
Science, Bogor Agricultural University, Bogor 16680, Indonesia. Authors are earnestly requested to
consult the “Instruction to Authors” published in Volume 2 Number 1. January 2011.

Indonesian Journal of Nutrition and Feed Science, published three times a year. AINI also conducted a
biannual National Seminar and Symposium in Nutrition and Feed Science
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

Indonesian Journal of Nutrition and

Feed Science
January 2011 Vol. 2 No. 1 : 1-45

CONTENTS

The Respond of Sago (Metroxylon sagu Rottb) Offered in the Ration Supplemented with
Probiotic on Production and Physical Quality of Duck’s Egg. A. A. A. S. Trisnadewi, T. G.
B. Yadnya ...................................................................................................................................... 1

Extracted Beta-Mannan from Copra Meal as an Alternative to Antibiotic Growth

Promotants in Broiler Diets. B. Sundu, E. Santo, L. Daisy, Damry, H. B ................................. 6

Growth Performance of Bali Cattle Bull (Bos sondaicus) Fed Fermented Cocoa
(Theobroma cacao L. ) Waste. I. M. Mastika, I. W. Supartha, I. W. Wiranatha, A. W. Puger .... 10

The Use of Cellulolytic Microbes from Cattle Rumen Fluid to Improve In vitro
Digestibilty of Fermented Robusta Coffee Pulp (Coffea canephora sp.). L. M. Yusiati, C.
Hanim, F. Az-zahra ...................................................................................................................... 15

Improving the Nutritive Value of Total Mixed Ration Based on By-products Fermented
by Rumen Liquor and Enzyme. I. M. Mudita, A. A. P. P. Wibawa, I. W. Wirawan, N. W.
Siti, I. G. L. O. Cakra .................................................................................................................... 20

The Evaluation of Nutrient Quality of Ramie Leaves Silage and Hay in Complete Mixed
Ration of Etawah-Crossbreed Goat Using in vitro Technique. Despal, I. M. L. Hutabarat, I.
G. Permana, R. Mutia.................................................................................................................... 26

Addition of Chlortetracycline to improve the Performance of broilers fed Local diets. R.

Murwani, I. A. Setyawan, A. K. Ariesta ....................................................................................... 32

Efficacy of Methionine Addition in Laying Hens Fed Corn Soy- Palm Kernel Meal Based
Diet. M. Ridla, Sumiati, J. Jachja, T. Toharmat, I. G. Permana and Nahrowi .............................. 36

The Effect of Earthworm Supplementation in the Ration on Growth Performance,

Carcass Production, and Abdominal Fat of Broiler. N. D. Dono, R. Damanik, J. Pasaribu,
A. Wibowo..................................................................................................................................... 42
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

REVIEWER ACKNOWLEDGMENT

Editorial team would like to thank the following scientists for their valuable contribution to the Indonesian Journal
of Nutrition and Feed Science, Vol. 2 No. 1, 2011.

Ali Agus Faculty of Animal Science, Gadjah Mada University, Indonesia

Arnold Sinurat Center for Research in Animal Science, Indonesia
A. R. Alimon University Putra Malaysia, Malaysia
Caribu Hadiprayitno Faculty of Animal Science, Jenderal Soedirman University, Indonesia
E. R. Ørskov Rowett Research Institute, England
Komang G. Wiryawan Faculty of Animal Science, Bogor Agricultural University, Indonesia
Nahrowi Faculty of Animal Science, Bogor Agricultural University, Indonesia
Ning Iriyanti Faculty of Animal Science, Jenderal Soedirman University, Indonesia
Osfar Sofjan Faculty of Animal Science, Brawijaya University, Indonesia
Suwarno Faculty of Animal Science, Jenderal Soedirman University, Indonesia
Sri Suharti Faculty of Animal Science, Bogor Agricultural University, Indonesia
Toto Toharmat Faculty of Animal Science, Bogor Agricultural University, Indonesia
Yantyati Widyastuti Research Center for Biotechnology, Indonesia
Indon. J. Nutr. and Feed Sci., January 2011, pages 1-5 Vol. 2 No. 1
ISSN 1410-2390

The Response of Sago (Metroxylon sagu Rottb) Offered in the Ration Supplemented
with Probiotic on Production and Physical Quality of Duck’s Egg

T. G. B. Yadnya * and A. A. A. S. Trisnadewi

Faculty of Animal Husbandry, Udayana University
Jl. PB Sudirman, Campus Bukit Jimbaran 80361 Denpasar, Bali-Indonesia
(Received 09-11-2009; Accepted 26-03-2010)

ABSTRACT

The research was carried out to study the response of sago (Metroxylon sagu Rottb)
inclusion in the ration which was supplemented with starnox to increase the production
and improve physical quality of duck’s egg. The experiment was designed using a
Completely Randomized Design (CRD) with three treatments consisted of : ration without
sago and starnox as control (C0), ration with 10% sago and 1% starnox supplementation
(C1), and ration with 20% sago and 1% starnox supplementation (C2). Each treatment
had five replicates with four ducks in each replicates. The data observed were feed
efficiency, egg production, and physical quality of duck’s egg. The results showed that
ration containing 10 - 20% sago could increase feed consumption and total egg weight
(P<0.05), and also improved feed conversion significantly (P<0.05) compared to that of
control treatment. The ration containing 10% and 20% sago increased hen day production
and the egg number significantly (P<0.05), meanwhile the average of egg weight was
not significantly (P>0.05) affected. However, the treatments had no significant effect
(p<0.05) on egg index, pH, specific gravity (P>0.05) but on the percentage of egg yolk
increased significantly (P<0.05). The physical composition of eggs with sago containing
ration showed that the eggshell percentage and egg yolk colour increased significantly
(P<0.05), and decreased the cholesterol (P<0.05) compared to that of control treatment.

Key words : Metroxylon sagu, starnox, production, physical quality, duck, egg

INTRODUCTION to be supplemented with probiotic, and one of them

The demand of maize increases as the source of
energy for human being, as well as feed for livestock,
so that maize needs to be substituted with appropriate
ingredients such as sago (Metroxylon sagu Rottb). The
nutrient of sago is almost the same as maize, but sago
has lower protein content which is only 5,28%, while
protein content of maize is 9,7% (Harsanto, 1992).
High fibre can reduce ration digestibility, so sago need
* Correspondence author
Faculty of Animal Husbandry, Udayana University
Jl. PB Sudirman, Campus Bukit Jimbaran 80361 Denpasar
Bali-Indonesia
e-mail: belawayadnya_fapet@yahoo.com
is starnox.
Starnox is a combination of starbio and pignox
(product of Medion, Bandung) contains enzymes,
and pignox contains minerals (Zn, Cu, Mn), vitamin,
antibiotic and methionin. Belawa and Suwidjayana
(2000) inclution of offer probiotic in the ration containly
high fibre (sawdust) improved the digestibility of
ration and fibre significantly. The starbio offered at
the ration containly different level of fibre increased
egg production significantly (Ariana, 2001). Starnox
offered in the ration increased egg production (Kenda,
2006). Belawa (2002) obtained that the ration offered
with different fibre sources and starbio reduced the
cholesterol content of duck’s eggs.
Sago offered as the substitution of maize at 0,
10, 20 and 30% combined with starnox on grower
phase did not affect the performance of duck at

January 2011 
YADNYA AND TRISNADEWI Indon. J. Nutr. and Feed Sci.

10 weeks of age (Witariadi et al., 2005). But the (C2). Each treatment consists of five replications
influence on egg production and egg quality had and each replication consists of four layer ducks.
not yet been studied, so further research is needed. The variables measured were as follows:
a. Feed consumption was ration given during
research minus residues of ration (g/head)
MATERIALS AND METHODS b. Total amount of egg was egg yielded
during research (egg/head during research)
The experiment was conducted in Guwang c. Average of egg weight was total egg weight
Village, Gianyar Regency in Bali for 12 weeks, divided by number of egg yielded during research.
while the dermination of the quality of duck egg d. Total egg weight was egg weight
was conducted in the Laboratory of Animal Products yielded during research (g)
Technology, Faculty of Animal Husbandry, Udayana e. Hen Day Production is number of egg
University for 4 weeks. yielded divided by number of duck
during research multiplied by 100%
Ducks
f. Physical quality of egg, included:
The ducks used in the experiment were 1. Egg index is egg length divided by width of the
layer ducks from UD Mertasari, Guwang egg
Village which were originally obtained 2. Physical composition was weight of each egg
from ducks breeder in Tabanan Regency. included eggshell, white and yolk of egg
divided by weight of egg and multiplied by
Ration and drinking Water 100
The rations were formulated based on 3. Colour of yolk was colour of egg yolk compared
Scott content (1982). The composition and to the rock yolk colour fan
nutrient of the rations are indicated in Table 1. 4. Egg acidity (pH) was measured with digital
pH
Table 1. The composition and nutrient content of the 5. Specific gravity of egg was determined by a
ration for ducks at the age 24 – 36 weeks hydrometer apparatus
6. Egg cholesterol by Liberman - Burchad was
Ingredients Treatment Rations
determined (Plummer, 1977).
(%) C0 C1 C2
The data were analyzed statistically with
Yellow maize 55.36 45.36 35.36
analysis of variance, and further analysis was
Sago - 10.00 20.00 continued using Duncan test (Steel and Torrie, 1989).
Fish meal 8.12 8.12 8.12
Rice bran 14.72 15.82 16.84
Copra meal 9.31 7.31 6.31 RESULTS AND DISCUSSION
Soybean meal 11.97 11.97 11.97
Feed conversion ratio (FCR)
Starnox - 1.0 1.0
NaCl. 0.52 0.42 0.40 The sago offered at the level of 20% in the
Nutrient composition ration increased feed consumption (Tabel 2). This
may be due to high fibre content in sago (21.68%)
ME (kcal/kg) 2879.6 2926.9 2831.9
(Harsanto, 1992) causing higher consumption
Crude protein (%) 19.3 18.36 17.89
to fulfill the energy requirement. Wahyu (1992)
Fibre (%) 4.64 5.77 6.72 reported that the first factor influencing consumption
Fat (%) 4.64 5.77 6.72 level in duck was the requirement of energy.
Calcium (%) 0.27 0.29 0.30 Starbio and pignox (starnox) added in the ration
Phosphor available (%) 0.39 0.33 0.31 improved the total egg weight yielded. The existence
of cellulase, pectinase, ligninase, protease and lipase
enzymes increased digestibility of the ration and also
Animal and treatments were allocated in a more amount of nutrient absorbed. Pignox consists
completely randomized design. The treatments consists of vitamins, minerals especially Zn assisting the
of ration without sago and starnox (C0), ration with metabolism, and also methionine amino acid useful
10% sago and supplemented with 1% starnox (C1), and in egg formation. Increasing the total egg weight
ration with 20% sago and supplemented with 1% starnox

 January 2011
Vol. 2 No. 1 THE RESPON OF SAGO

caused the feed conversion ratio more efficient. In
fact, the result indicated that 10 - 20% of sago offered
with starnox supplementation had lower FCR values
than the control treatment. Kenda (2006) reported
starnox in the ration containing different fibre content
improved total egg weight and more efficient in ration
utilization compared to that in control treatment.
Egg production
Sago offered in the ration supplemented with
starnox increased egg production, both for total egg
number and total egg weight (Table 3). Ariana (2001)
obtained that starbio addition in the ration containing
different fibre increased egg production. Kenda (2006)
reported that using starbio and pignox in the ration
improved number of eggs and hen day production. The
average egg weight was not significantly improved
(P>0.05), this might be caused by insufficiency of
supply nutrients. Ariana (2001) obtained that starbio
addition in the ration containing different fibre
increased egg production. Kenda (2006) reported that
using starbio and pignox in the ration improved number

Table 2. The effect of sago (Metroxylon sagu Rottb) offered in the ration supplemented with starnox on feed conversion ratio
and total eggs weight at the first layer phase

Treatments
Variables SEM
C0 CI C2
Feed Consumption (g/duck) 12743.30b 12900.16b 13657.86a 176.43
Total Egg Weight (g/duck) 2790.21b 3079.52a 3104.78a 45.34
FCR 4.60a 4.18b 4.40c 0.05
Note: C0 : Ration without sago and starnox; C1 : Ration with 10% sago and 1% starnox; C2: Ration with 20% sago and 1% starnox;
Means in the same row with different superscript differ significantly (P<0.05) ; SEM : standard error of the treatment means.

Table 3. The effect of sago (Metroxylon sagu Rottb) offered in the ration supplemented with Starnox on bali duck egg
production at the first layer phase
Treatment
Variables SEM
C0 C1 C2
Hen Day Production (%) 53.88b 59.83a 60.23a 0.85
Total Number of Egg (egg/head) 45.25b 50.25a 50.59a 0.70
Average Egg Weight (g/egg)ns 61.05 61.22 61.37 0.19
Note: C0 : Ration without sago and starnox; C1 : Ration with 10% sago and 1% starnox; C2: Ration with 20% sago and 1% starnox;
Means in the same row with different superscript differ significantly (P <0.05); SEM : standard error of the treatment means.

of eggs and hen day production. The average egg
weight was not significantly improved (P>0.05), this
might be caused by insufficiency of supply nutrients.
Physical quality of eggs
Egg index is comparison between length
and width of the eggs. If the egg index value
close to one, it means that the egg closely become
circular. The result showed that egg index was not
significantly different, this possibly because of the
egg weight average was not significantly different,
so the size of eggs were the same. Nutrient needed
by duck in the ration have been fulfilled, and did
not affect the length and width of the eggs (Table 4).
Sago offered in the ration supplemented with
starnox improved the colour of yolk. This might be
related to the addition pignox that containing nutrients
which are deposited in the yolk. Pignox consists of
vitamins soluble in fat (A, D, E, K), vitamin B-complex
(Bl, B2, B6, and B12), also the micro minerals (Mn,
I, Fe, Co, Cu, Zn) and methionine amino acid. The
nutrients in pignox have similar nutrients composition
found in the yolk colour. The acidity degree (pH) of
egg was not significantly different. This is possibly
because of the same egg weight and specific gravity
of egg, so the content of ion H+ in the egg is equal.
Sago offered in the ration supplemented with
starnox improved the yolk and egg shell percentage,
while decreased the percentage of the white egg
percentage. It is proven that the ration containing

January 2011 
YADNYA AND TRISNADEWI Indon. J. Nutr. and Feed Sci.

Table 4. The effect of sago (Metroxylon sagu Rottb) offered in the ration with starnox supplementation to physical quality
of Bali duck egg at the first layer phase
Treatments
Variables SEM
C0 C1 C2
Egg index ns 0.68 0.60 0.70 0.39
Colour of yolk 6.90b 7.46ab 7.8a 0.19
Acidity (pH) ns 6.6 7.06 7.12 0.10
Egg specific gravity ns 1.070 1.072 1.078 1.19
Egg physical composition (%)
Eggshell ns 10.53 11.26 12.13 0.42
Albumin 55.55 a
53.35 b
52.28 b
0.48
Yolk 33.92b 35.39a 35.59a 0.35
Egg cholesterol 570.29 a
411.34 b
418.45 b
22.45
Note: C0 : Ration without sago and starnox; C1 : Ration with 10% sago and 1% starnox; C2: Ration with 20% sago and 1% starnox;
Means in the same row with different superscript differ significantly (P<0.05); SEM : standard error of the treatment means.

sago and starnox increased the specific gravity or
eggshell percentage. Starbio improved digestibility
of ration, so higher amount of nutrients absorbed,
and pignox consists of nutrient needed for yolk
formation such as : Mn, I, Fe, Co, Cu, Fe. Higher
feed consumption could cause higher value of specific
gravity and eggshell than the control treatment (C0).
Sago offered as a fibre source decreased
egg cholesterol. Fibre content in C1 in the form
of glucosides bound cholesterol, so reduced the
cholesterol absorbed or accumulated in egg (Alan
et al. 1976, In Budaarsa, 1997). Besides, in large
intestine the fibre was fermented into propionic acid
which affect the cholesterol formation in the liver,
through formation of 3 methyl, 3 hydroxy glutenyl,
and -CoA reductase enzyme. Cholesterol formation
in the liver decrease, meaning that the cholesterol
circulation decreased and finally less cholesterol
accumulation in eggs. Bestari et al. (1984) obtained
that fibrous ration reduced the cholesterol content in
eggs. Belawa (2002) reported that starbio offered in
fibrous ration reduce the cholesterol content in eggs.
CONCLUSION
Based on the result of the research, it could be
concluded as follows:
1. Ration containing 10 - 20% sago which was
supplemented with starnox could improve feed
conversion ratio and egg production of Bali duck
at first layer phase.
2. Ration containing 10 - 20% sago which was
supplemented with starnox could improve the
physical quality of egg, especially increased
the percentage and colour of egg yolk and also
decreased the cholesterol content in eggs.
ACKNOWLEDGEMENT
Thanks due to the Rector of Udayana University
through the Head of Research Institute of Udayana
University which provided fund (DIPA) for budget
year 2008, so that the research could be conducted.
REFERENCES
Ariana, I N. T., 2001. Pengaruh Tingkat Serat yang Berbeda
dalam Ransum yang disuplementasi dengan Strabio Plus
terhadap Produksi Telur Itik Bali. Laporan Penelitian.
Fakultas Peternakan Universitas Udayana, Denpasar.
Belawa, Y. T. G. dan I N. Suwidjayana. 2000. Respon
Pemberian Effective Microorganism-4 (EM-4) dalam
Pakan Berserat terhadap Daya Cerna Ransum pada
Ayam Pedaging. Prosiding Seminar Nasional, BPTP
Bali.
Belawa, Y. T. G. 2002. Pengaruh Pemberian Tingkat Serat
Berbeda yang Disuplementasi dengan Probiotik
Starbio terhadap Konversi Ransum, Kadar Protein,
Asam Urat, Kolesterol Darah dan Komposisi Karkas
Itik Bali Umur 10 Minggu. Proc. Seminar Nasional.
IP2TP, Denpasar.
Bestari, A. P., Suerat, A. D. Setioko, F. Setiadi dan Ulupi.
1984. Pengaruh Berbagai Tingkat Serat Kasar Dalam
Ransum terhadap Produksi dan Kualitas Telur Itik
Tegal. Proc. Agro-Industri, Peternakan di Pedesaan.
Balai Penelitian Ternak, Ciawi Bogor. Hal. 120-127

 January 2011
Vol. 2 No. 1
Budaarsa, I . K. 1997. Kajian Penggunaan Rumput Laut
dan Sekam Padi untuk Menurunkan Kadar Lemak dan
Kolesterol Daging Babi. Disertasi S3. IPB Bogor.
Harsanto. P. B. 1992. Budidaya Tanaman dan Pengelolaan
Sagu. Cetakan Ketiga, Kanisius. Yogyakarta .
Kenda, Y. 2006. Pengaruh Pemberian Starbio dan Starnox
dalam Ransum terhadap Efisiensi Penggunaan
Ransum pada Itik Bali Fase Peneluran Pertama (24-
32 minggu). Skripsi. Jurusan Peternakan, Fakultas
Pertanian Univeristas Warmadewa, Denpasar.
Plummer, D. T. 1977. An Introduction to Practical
Biochemistry. McGraw-Hill Book Co. Ltd. New
Delhi.
Scott, M. L., M. C. Neisheim and R. J. Young. 1982. Nutrition
of the Chicken. 2nd Ed. M.L. Scott and Assoc. Ithaca,
New York.
THE RESPON OF SAGO
Steel, R. G. D. And J. H. Torrie. 1989. Principles and
Procedure of Statistics. McGraw Hill Book Co. Inc.
New York.
Wahyu, J. 1992. Ilmu Nutrisi Unggas. Cetakan ke-3. Gadjah
Mada University Press. Yogyakarta.
Witariadi, N. M., N. G. K. Roni, I G. L. O. Cakra. 2005.
Evaluasi terhadap Substitusi Jagung dengan Sagu
Cincang dalam Ransum yang disuplementasi ”
Starnox” pada Itik Bali Awal Periode Pertumbuhan.
Laporan Penelitian Fakultas Peternakan Universitas
Udayana Denpasar.
January 2011 
Indon. J. Nutr. and Feed Sci., Januari 2011, pages 6-9 Vol. 2 No. 1
ISSN 1410-2390

Extracted Beta-Mannan from Copra Meal as an Alternative to Antibiotic Growth

Promotants in Broiler Diets

B. Sundu a*; E. Santo b, L. Daisy b and Damry H.B a

a
Department of Agriculture, Faculty of Animal Husbandry, University of Tadulako, Palu, Indonesia
b
Agricultural Beureau, Jl. R. A Kartini No. 80, Palu, Indonesia
(Received 09-11-2009 ; Accepted 20-08-2010)

Abstract

An experiment was conducted to determine the prebiotic function of extracted beta mannan
from copra meal in an attempt to replace antibiotic growth promotants. A total of two
hundred DOC (day old chicks) were used and the birds were distributed into 6 treatment
diets. The birds were given starter diets from day 1-21 and grower diets from day 22-42. The
treatment diets were: (1) control diet, (2) control diet + avilamycin, (3) control diet + 0.025%
copra mannan, (4) control diet + 0.05% copra mannan, (5) control diet + 0.075% copra mannan
and (6) control diet + commercial mannan (yeast mannan). Feeds were offered ad libitum
and water was available at all times. A completely randomized design was used with 6
treatment diets and 5 replicate cages. Differences among treatments were tested by analysis
of variance and then Duncan Multiple Range Test. Data indicated that supplementation
of antibiotic (avilamycin), 0.05% copra mannan and 0.02 % commercial mannan increased
(P<0.05) body weight gain. Feed conversion ratio, feed intake, proportion of gram negative
bacteria, mortality, intestinal dimension were not affected by antibiotic and mannan.
Caeca pH of birds fed the commercial mannan in the diet was lower than those of birds
fed the control diet. In conclusion, 0.05% copra mannan can be used to replace antibiotic
(avilamycin) and the body weight of birds fed the 0.05% copra mannan increased to the
same level of the body weight of birds fed the diet either supplemented with antibiotic or
commercial mannan.

Key words : Beta mannan, copra meal, antibiotic, broiler

Introduction been banned in European Union since January 2006.

The public is placing some blame of antibiotic
resistance of some potential pathogen bacteria due
to wide use of antibiotic growth promotants in
poultry. Debate over resistance found among some
negative bacteria, such as E. coli and Salmonella, has
generated objection to antibiotic use. Therefore, the
use of antibiotic growth promotants in broiler diets has
*Correspondence author
Animal Husbandry Department, Agriculture Faculty
University of Tadulako
E-mail : b_sundu@yahoo.com
Glucomannan (mannan oligosaccharides) from yeast
(Saccharomyces cereviciae) has been commercially
introduced as a feed additive for poultry since 2003.
The capacity of mannan oligosccaharydes to block
the colonization of pathogenic bacteria in the intestine
of broilers resulted in an increased body weight gain
and a decreased population of intestinal pathogenic
bacteria and mortality (Lyons, 2002).
The efficacy of using beta mannan from palm
kernel meal as an alternative to antibiotic growth
promotants has been well reviewed by Sundu et al.
(2006). Copra meal containing 30-40% beta mannan
may have properties, in common, with the mannan
from palm kernel meal or from yeast (Sundu and

 January 2011
SUNDU ET AL. Indon. J. Nutr. and Feed Sci.

Table 1. Nutrient composition of the experimental diet A method of Kusakabe and Takashi (1988) was
Ingredients Starter diet Grower diet used to extract beta mannan from copra meal. A 24
(%) (%) liters of 24% NaOH was added to 3 kg in a stainless
steel bucket. The mixture was occassionally stirred for
Soybean meal 25.00 24.00
24 hours at room temperature. The slurry is filtered
Corn 57.80 59.40 through a cloth bag. The filtrate was neutralized with 12
Fish meal 12.00 10.00 NH2SO4 until the pH solution is about 5.5. Resultants
Rice bran 3.00 6.00 precipitate (copra mannan) collected by centrifugation,
Dicalcium phosphate 1.50 0.9 was dialysed against tap water to remove salts.
Detection of gram negative bacteria was done at
Salt 0.30 0.66
the laboratory of plant diseases at Tadulako University.
Methionine 0.10 0.55 To detect gram-negative bacteria, the method of gram
Lysine 0.10 0.11 stain was used based on the procedure of Tortora et
Premix 0.20 0.20 al. (1982). The heat-fixed smear was covered with
a basic purple dye. After a short time, the dye was
Crude protein 23.13 21.00
washed off and the smear was covered with iodine.
Crude fibre 3.5 3.6
When the iodine was washed off, both gram positive
ME (MJ/kg) 13.39 13.39 and negative bacteria appear dark violet. The slide was
Lysine 1.1 1.0 then washed off with an ethyl alcohol solution. This
Methionine + cystiene 0.9 0.79 solution acted as a decolourizing agent, removing the
purple colour from the cells. The alcohol was rinsed
and the slide was stained with safranin. The purple dye
Table 2. Experimental diets and the iodine combine with the bacteria and colour
Diet Composition was dark violet. Bacteria that retain this colour after
R1 Basal diet attempted decolourization were classified as gram
positive bacteria (dark violet). Bacteria that was
R2 Basal diet + antibiotic (avilamycin) (6 ppm)
invisible (colourless) was gram negative. Application
R3 Basal diet + 0.025% extracted copra mannan of safranin caused the gram negative bacteria to turn
R4 Basal diet + 0.050% extracted copra mannan pink.
R5 Basal diet + 0.075% extracted copra mannan A completely randomized design was used
with 6 treatment diets and 5 replicate cages. The
R6 Basal diet + 0.020% commercial from yeast treatments diets were: (1) control diet, (2) control diet
+ avilamycin, (3) control diet + 0.025% copra mannan,
(4) control diet + 0.05% copra mannan, (5) control
Dingle, 2003). This experiment, therefore, aimed to diet + 0.075% copra mannan and (6) control diet +
examine the potential use of extracted beta mannan commercial mannan (yeast mannan).
from copra meal as a feed additive in broiler diets.
Data were analyzed of variance. Differences
among treatments were tested by analysis and then
Materials and Methods Duncan Multiple Range Test (Steel and Torrie, 1980).

The study was conducted in the poultry house

at the University of Tadulako Palu, Indonesia. Two
hundred day old chicks were used as experimental
animals. They were placed in brooder cages for two
weeks. The birds were, then, transferred to the floor
cages.
From day 1-21, the birds were fed starter diets and
grower diets were offered from day 22 -42. Feed and
water were given ad-libitum throughout the trial. To
avoid the use of unnecessary antibiotic, antibiotic-
containing feed additives were not applied in this
experiment. Feeds were offered ad libitum and water
was available at all times.
Results and Discussion
The means of body weight gain, feed intake,
feed conversion ratio (FCR), mortality, intestinal
dimensions, caeca pH and proportion of gram negative
bacteria are shown in Tables 3 and 4. Feed intake, FCR,
mortality, proportion of gram negative bacteria and
intestinal dimesions were not affected by treatments
(P>0.05) while the effects of treatments on body
weight gain and caeca pH were significant (P<0.05).
Supplementation of diet with either 6 ppm avilamycin,
0.05% extracted copra mannan or 0.02% commercial

January 2011 
Vol. 2 No. 1 EXTRACTED BETA -MANNAN

Table 3. The effect of extracted copra mannan on the response of broiler chickens.
Treatments Body weight Feed intake FCR Mortality
gain (g) (head)
Control diet (CD) 1501b 3053 2.03 2
CD + avilamycin 1650a 3075 1.86 2
CD + 0.025% extracted copra mannan 1625 ab
2834 1.74 2
CD + 0.050% extracted copra mannan 1701 a
2809 1.65 3
CD + 0.075% extracted copra mannan 1544 b
2876 1.86 2
CD + 0.020% yeast mannan commercial 1705 a
2794 1.64 2
Note : Means in the same column with different superscript differ significantly (P < 0.05)

Table 4. Response of digestive tract of broilers fed extracted copra mannan

Treatments Length of intestine Weight of intestine pH Proportion of
(cm/kg BW) (g/kg BW) caeca negative gram
bacteria
(%)

Control diet (CD) 132.2 22.6 7.3a 24.0

CD + avilamycin 137.9 32.4 7.1 ab
25.8
CD + 0.025% extracted copra mannan 142.7 26.6 7.1ab 16.4
CD + 0.050% extracted copra mannan 127.9 37.3 6.9ab 20.0
CD + 0.075% extracted copra mannan 140.8 39.1 6.8ab 20.4
CD + 0.020% yeast mannan commercial 123.5 32.5 6.7b 20.0
Note : Means in the same column with different superscript differ significantly (P <0.05)

yeast mannan had higher body weight gain than those
of birds fed the control basal diet. Caeca pH of birds
fed the diet supplemented with commercial yeast
mannan was significantly higher (P<0.05) than the pH
caeca of birds fed the control diet.
The use of antibiotic (Evangelisti et al., 1975) to
improve bird performance has long been believed. A
positif effect of using antibiotic, in this current study,
was an increased body weight gain from 1501 g to
1650 g. This improvement may be due to improved gut
health. Related to a decreased in bacterial population in
the caeca of birds fed the diet containing antibiotic. It
is not hard to razionalise this fenomenon as antibiotic
have a killing properties againts bacteria. This
condition could minimize negative effect of pathogen
bacteria.
Improvement of live weight gain was also found in
birds fed the commercial yeast mannan. These results
were supported by the previous findings of Shane
(2006). Birds fed the diet supplemented with 0.05%
extracted copra mannan could reach growth equal to
that of chickens fed the commercial yeast mannan.
This finding may justify that copra mannan at a right
concentration could have a prebiotic properties. It is
hard to elaborate why concentration below or above
0.05% did not work well on this experimental diet.
Caeca pH of birds fed commercial mannan
was lower than those of birds fed the control diet.
Interestingly, caeca pH of birds fed extracted copra
mannan tended to linearly decrease over an increased
concentration of copra mannan in the diet. The
mechanism that produces the decreased pH due to
mannan supplementation is still unclear. It may be
through mode of action in which mannan, either from
yeast or copra meal, are fermented in the caeca as a
results of indigestibility of this fraction in the digestive
tract of broiler. One of the product of fermentation
is lactic acid (Wang and Gibson, 1993; Okumura et
al., 1994), which prevents the growth of pathogenic
bacteria such as E. coli and Salmonella (gram negative
bacteria). This may be the reason of a small decrease
in proportion of gram negative bacteria. In conclusion,
extracted copra mannan could be used at 0.05% to
replace antibiotic growth promotant. The efficacy of
copra mannan was equal to commercial mannan.

 January 2011
SUNDU ET AL.
Conclusion
Extracted copra mannan could be used at 0.05%
to replace antibiotic growth promotant. The efficacy of
copra mannan was equal to commercial mannan.
References
Evangelisti, D.G., A.R. English, A.E. Girard, J.E. Lynch &
L.A. Solomons. 1975. Influence of subtherapic levels
of oxytetracycline on Salmonella typhimirium in
swine, calves and chickens. Antimicrobial Agents and
Chemotherapy, 8: 664-672.
Kusakabe, I. & R. Takashi. 1988. Enzymatic preparation
beta 1-4 mannooligosaccharides and beta 1-4 gluco-
mannooligosaccharides. Methods in Enzymology,
160: 518-523.
Lyons, T. P. 2002. Navigating from niche markets to
mainstream: A feed industry Kakumei. Proceedings
of Alltech’s 16th Anuual Asia Pacific Lecture Tour, pp:
1-16.
Okumura, J., M. Furuse, T. Kawamura, K. Toyoshima,
M. Sugawara, T. Suzuki, G. Seo & Soga, H. 1994.
Effects of glucooligosaccharides and bacteria on egg
production rate and caecal bacteria population in
the chicken. Japanese Poultry Science, 31: 189-194.
Indon. J. Nutr. and Feed Sci.
Shane, S. M. 2006. Mannan oligosaccharide in poultry
nutrition. Mechanisms and benefits. Nottingham
University Press. UK.
Sundu, B., A. Kumar & J. Dingle. 2006. Palm kernel meal
in broiler diets: its effect on chicken performance and
health. World’s Poultry Science Journal. 62: 316-325.
Sundu, B. & J. Dingle .2003. Use of enymes to improve
the nutritional value of palm kernel meal and copra
meal. Queensland Poultry Science Symposium, 11,
(14): 1-15.
Steel, R. G .D. & J. A. Torrie.1980. Principles and procedures
of statistics. New York, McGraw Hill.
Tortora, G. J., B. R. Funke & C. L. Case. 1982. Microbiology,
an introduction. Cummings publishing company,
Menlo Park, California.
Wang, X. & G.R. Gibson, 1993. Effects of in vitro
fermentation of oligofructosa and inulin by bacteria
growing in the human large intestine. Journal of
Applied Bacteriology, 75: 373-380
January 2011 
Indon. J. Nutr. and Feed Sci., January 2011, pages 10-14 Vol. 2 No.1
ISSN 1410-2390

Growth Performance of Bali Cattle Bull (Bos sondaicus) Fed Fermented Cocoa
(Theobroma cacao L.) Waste

I M. Mastika a, I.W. Supartha b , I.W. Wiranatha c, dan A.W. Puger a*

Faculty of Animal Husbandry, University of Udayana
a

Jl. PB Sudirman Denpasar Kampus Bukit Jimbaran 80361, Bali-Indonesia

b
Faculty of Agriculture, University of Udayana
Jl. PB Sudirman Denpasar Kampus Bukit Jimbaran 80361, Bali-Indonesia
c
Livestock Services of Tabanan Regency, Bali, Indonesia
Br. Dauh Pala, Ds. Dauh Peken, Jln. Kutilang No. 6
(Received 09-11-2009 ; Accepted 10-05-2010)

ABSTRACT

An experiment was carried out to study the growth performance of Bali cattle (Bos sondaicus)
bull fed fermented cocoa waste. A total of 16 Bali cattle with the range of body weight 220-
225 kg were divided into two groups of 8 cattle each following the pairing method. The
first group was given 2 kg head/day of fermented cocoa by product and the second was
given 2 kg/head/day of commercial feed. Native grass and water were provided ad libitum.
This experiment was run for 12 weeks. Final body weight of the cattle were 257.5 kg and
263 kg respectively for those given 2 kg fermented cocoa by product and 2 kg commercial
feed (P>0.05). Daily weight gain of the cattle fed fermented cocoa waste and commercial
feed were 451 g/day and 488 g/day respectively (P>0.05). Total intake dry matter recorded
during the experiment were 595.61 kg and 582.32 kg for those given fermented cocoa
waste and commercial feed (P<0.05). However, there is no significant different on the feed
conversion ratio for both groups. Digestibility coefficient of crude fibre, dry matter, organic
matter, crude protein, crude fat and NFE were not significant different for both groups. It
was concluded that fermented cocoa waste has a prospective future for ruminant feed in
Indonesia.

Key words : Bali cattle, fermented cocoa by product, digestibility, feed intake

INTRODUCTION species. In tropical country like Indonesia, the day

The constan increase of population in Indonesia
result in the productive land has been transformed
to resettlements and this condition reducing to some
extend the forage availability for animal feeds. This
situation will become a mayor constraint in the future
in increasing animal production especially ruminant
* Correspondence author
Fakultas Peternakan Unud, Jl. PB. Sudirman Dps Bali
Telp: (0361)222096
E-mail: puger_anton@yahoo.com
length or the climate, soil condition will affect also
the quality and quantity of the forage production,
although some effort have been made to improve
pasture production by introducing new species of
grass and legume but the degree of success varies
from one to another place. To overcome this problem,
the possibility of using agriculture and industrial
agriculture by product must be considered as one
among those possible alternatives in feeding animals.
Cocoa (Theobroma cacao L) by product is one of the
estate plantation which is potentially producing waste
which can be used as animal feeds (Laconi, 1998 and
Mastika, 2006).
Cocoa plantation in Indonesia in general and in
Bali especially, has increased continously by 7% per

10 January 2011
MASTIKA ET AL.
year. in 2006, the plantation area of cocoa in Bali
was 11,154 ha. The Jembrana and Tabanan regency
are considered as the centre of cocoa production, and
the total seed production estimated 2.62 ton/ha/year
(Disbun Bali, 2006). The ratio is of seed and cocoa
shell is 1: 3.4 in fresh materials (Ginting, 2004). Based
on the figure it was calculated that the cocoa waste
availability in Bali was 8.91 ton/ha/year.
The nutrient content of cocoa shell was reported
by Suharto (2004). The crude protein, fat, crude fiber
and total digestible nutrient (TDN) was 9.15%,
1.25%, 32.7% and 50.3% respectively. Although the
nutrient content is good for animal feed, however, the
material contains some limiting factor such as tannin,
theobromine and high crude fiber content as considered
as a toxic factors (Ginting, 2004). Theobromine and
tannin substances has a high affinity to protein and
carbohydrate causing these two nutrients decreased
in their availability (Amirroenas, 2005) and finally
affecting the growth of rumen microorganisms which
is finally responsible for animal growth. Agricultural
by product quality can be done improved using
biofermentation process (Purnama, 2004). The
common biofermentation agent is Aspergillus niger
and the fermentation process of cocoa shell decrease
theobromine and tannin content up to 80-83% and 91-
98% respectively (Guntoro, 2006), and significantly
increase protein content (91.98%) from 8% to 15.3%
crude protein (Purnama, 2004).
Cocoa shell is a high potential of animal feed due
to the high content availability of its nutrient content,
however, since cocoa shell is known containing
theobromine and tannin, it is worth therefore to run
the experiment using A. niger to ferment cocoa shell
as cattle feed.
MATERIALS AND METHODS
Animals
A total of 16 Bali cattle bull with the average
weight of 220 kg, age approximatelly 2 years were
used in this experiment. Animals were divided into
two treatment groups of 8 animals each as replicates.
Feeds and water
As basal feeds, all animals were given native
grass and each group was supplemented with either 2
kg commercial feed per head per day as treatment A,
or 2 kg of ground fermented cocoa shell as treatment
B. Fresh native grass and water were provided ad
libitum.
Indon. J. Nutr. and Feed Sci.
Management and Housing
Animals were placed in individual pen in one shed
and the size of each pen was 1.5 m x 2 m, and each of
them was provided separate feeding throughs either for
grass or concentrate. The size of feeding through for
grass was 0.8 x 0.5 x 0.5 m and for concentrate was 0.4
x 0.5 x 0.5 m. A 20 litres plastic bucket is also provided
for drinking water. Before the commencement of
the experiment, all animals were vaccinated againt SE
and dewormed.
Cocoa shell for fermentation
Fermenting agent used in this biofermentation
was A. niger provided by BPTP Bali Province. The
preparation of fermentation followed the method
described by Guntoro (2006). One kg of sugarcane,
1 kg fertilizer (NPK), 1 kg of urea and 1 litre of
concentrated seed of A. niger were mixed in 100 lt of
clean fresh water. This mixture was kept in the room
temperature for 3 days before used. The fresh cocoa
shell was cut into 2-3 cm pieces and 1,000 kg of these
fresh chopped cocoa shell were mixed with 100 litres
A. niger solution and then was placed in a fermenting
box. Fermentation process was run for 5-6 days then
the fermented cocoa shell was dried under the sunshine
for 2-3 days. The dry fermented cocoa shell were
ground using grinding machine (chopper; Kimdong
185 Type made in China), to make a powder.
In this experiment a pairing method was used,
consisted of two treatments and 8 replicates. Animal
in the first group were offered native grass + 2 kg
commercial feed as treatment A and the second group
were given fresh grass + 2 kg of fermented cocoa as
treatment B. This experiment was run for 12 weeks.
Body weight and daily weight gain, feed consumption,
feed conversion ratio were used as parameters of the
treatment effects.
At the end of the experiment, all data collected
were analyzed using Pairing Test (t-test) as explained
by Chang (1979).
RESULTS AND DISCUSSION
Data of laboratory analysis showed that there
were quite significant changes of the nutrients and
antinutrients content of cocoa shell fermented with
A. niger compared to those non fermented materials
(Table 1).
The protein content of the feed stuffs increased
by 102.18% and on the other hand the crude fiber
content decreased 7.05% compared to unfermented
cocoa shell. This finding is in line with the results
January 2011 11
Vol. 2 No. 1 GROWTH PERFORMANCE

Table 1. Nutrient content of commercial feed, fermented cocoa, and unfermented cocoa by product.
Feedstuffs
Nutrients/ Antinutrients Unfermented cocoa
Commercial feed Fermented cocoa
by product
Nutrients
Dry matter % 92.750 88.080 88.260
Crude protein % 9.720 13.870 6.860
Crude Fiber % 21.625 25.820 27.780
Fat % 2.599 5.190 4.770
Total Digestible Nutrient % 58.134 49.580 21.030
Nitrogen Free Extract % 54.337 43.306 39.980
Digestible energy Mcal/kg 2.563 2.186 -
Ash % 11.510 11.120 8.870
Antinutrients
Theobromine % - 3.460 5.110
Cyanide ppm - 91.130 132.980
Source : Data of proximate analysis from Nutrition Laboratory, Faculty of Animal Husbandry, Udayana University

reported by Guntoro et al. (2002) and Guntoro (2006).
It was postulated that biofermentation using A. niger
could significantly improve the quality of the cocoa by
product as an alternative feedstuff for animals.
Table 2 shows that when this fermented cocoa shell
was given to Bali cattle at the rate of 2 kg/head/day,
the animals grew as good as those given commercial
feed supplementation (P>0.05) (treatment A) that
was 257.50 kg vs 263.0 kg. The same trend in body
weight gain and daily weight gain were performed
by the animals fed either fermented cocoa shell or
commercial diet supplementation.
However, feed intake of the cattle fed fermented
cocoa feed significantly (P<0.05) higher than that
given commercial concentrate (Table 2), on the other
hand energy intake and feed conversion ratio of both
treatments were not significant. The lower feed and
energy was apparently responsible for the slightly
lower body weight gain of the cattle fed fermented
cocoa feeds. As stated by Parrakasi (1999) and Ginting
(2004), that feed consumption of the ruminant basically
to statisfy their energy requirement, thus the animals
will stop eating whenever their energy requirement is
satisfied or fulfilled.
The hemicellulose content as neutral detergent
fiber (NDF) fraction of the cocoa shell is high, but
most of it is bound to tannin or theobromine causing
this materials have a low or can not be digested in the
rumen (Amirroenas, 2005). Biofermentation using A.
niger causing the tannin and theobromine content were
reduced significantly (Purnama, 2004; Guntoro 2006).
They postulated that decreasing the content of both
tannin and theobromine and finally the availability
of the hemicellulose that can be digested will be
increased. Probably, the higher rate of hemicellulose
digestion in animals given fermented cocoa shell
(treatment B) causing higher crude fiber digestibility
(Table 2) and stimulate higher feed consumption on
fermented cocoa fed animals. This finding supported
by the report of Preston (1986) that the more accessible
of hemicellulose to microorganism in the rumen will
therefore increase total fermentation and the rate of
fermentation. The interesting point in this experiment
was that the feed conversion ratio of the cattle fed
commercial feed vs fermented cocoa shell was
slightly higher on treatment B although statistically
not significant. Probably this differences due to the
fact that the NFE consumption of cattle on treatment
B was lower than cattle on treatment A. Other factor
that could be responsible for a lower efficiency of
fermented cocoa fed animals is probably the tannin
and theobromine content is not completely discarded
during biofermentation process (Table 1).
From the present results, it was suggested that
cocoa shell waste has a potential role for animal feed
in the future. With the technology development,
the quality of this feedstuff can be improved by
physical, chemical and biofermentation process.
Supplementation of 2 kg/head/day of fermented cocoa
gave no significant different (451 g/head/day vs 488

12 January 2011
MASTIKA ET AL. Indon. J. Nutr. and Feed Sci.

Table 2. Body weight, body weight gain, feed intake, feed conversion ratio of bali cattle bull offered 2 kg commercial feed or
2 kg fermentation cocoa shell supplementation/head/day
Treatments
Parameters SEM
A B
Body Weight
Initial body weight (kg) 222.00 219.63 5.35
Final body weight (kg) 263.00 257.50 4.87
Body weight gain (kg) 41.00 37.88 2.16
Daily body weight gain (g/head/day) 488.00a 451.00a 25.76
Dry Matter Intakes
Total grass (kg) 426.67a 450.81b 3.16
Total Feeds (kg) 582.32a 595.61b 3.11
Daily grass (kg/head/day) 5.11a 5.37b 0.04
Daily Feeds (kg/head/day) 6.93a 7.09b 0.04
Feed energy consumption (Mcal DE/head/day) 17.21 16.87 16.26
Feed Conversion Ratio (FCR) 14.78 16.32 1.00
Digestibility Coefficient (%)
Dry matter 62.22 60.59 1.80
Organic matter 64.48 64.54 0.68
Crude protein 69.28 68.95 2.34
Crude fiber 57.37 63.12 1.87
Crude fat 67.61 54.85 5.21
NFE 70.42 64.85 1.62
Note : A : fresh native grass fed ad libitum + 2 kg commercial feed; B : fresh native grass ad libitum + 2 kg fermented cocoa shell;
SEM = standard error of the treatment means; Means in the same row with different superscript differ significantly (P <0.05)..

g/head/day) in gain compared to those given 2 kg
commercial feed supplementation and this daily gain
is almost double compared to animal fed grass only
which was only 200 g-300 g/head/day as reported by
Sukanten et al., (1990). The overall finding is that the
supplementation of fermented cocoa shell will reduce
feed cost in animal production.
group association who help in feeding and looking after
the animals during the period of experimentation. The
Laboratory Staffs of the Faculty of Animal Husbandry
for the assistance in analysing the nutrient content of
the feedstuffs are also acknowledged.
REFERENCES

CONCLUSION Amirroenas, D.E. 1990. Mutu Ransum Berbentuk Pellet

It was concluded that fermentation of cocoa shell
with A. niger increased nutrients content and decreased
theobromine and cyanide of cocoa shell. Daily gain,
feed conversion ratio of Bali cattle fed 2 kg/head/day
of fermented cocoa shell were similar to those given
of commercial feed. The overall finding suggested
that the supplementation of fermented cocoa shell
will reduce feed cost and has a prospective future for
ruminant production.
ACKNOWLEDGEMENT
The authors wish to thanks all staffs of the
Goverment of Estate Province of Bali for fund support
in this research. Thanks are also due to the farmers
dengan Bahan Serat Pod Coklat (Theobroma cacao
L) untuk Pertumbuhan Sapi Perah Jantan. Thesis
Magister. Fakultas Pascasarjana, IPB.
Chang, L. C. 1979. The Concept of Statistics in Connection
with Experimentation . Extention Bulletin No 13.
Food and Fertilizer Technology Center. Taiwan.
Dinas Perkebunan Provinsi Bali. 2006. Demoplot
Pengendalian Penggerek Buah Kakao dengan
Pola Integrasi. Laporan DISBUN Provinsi Bali
bekerjasama dengan Fakultas Pertanian Unud.
Dinas Peternakan Provinsi Bali. 2006. Data Populasi
Peternakan di Provinsi Bali Tahun 2002 sampai dengan
2006. Dinas Peternakan Provinsi Bali. Denpasar.
Ginting, S.P. 2004. Tantangan dan Peluang Pemanfaatan
Pakan Lokal untuk Pengembangan Peternakan

January 2011 13
Vol. 2 No. 1
Kambing di Indonesia. Disampaikan pada Lokakarya
Nasional Kambing Potong. Pusat Penelitian dan
Pengembangan Peternakan. http://peternakan.
litbang. deptan. go. id. [30 Januari 2007].
Guntoro, S. 2006. Mengolah Limbah Perkebunan untuk
Pakan Ternak. Balai Pengkajian Teknologi Pertanian
(BPTP) Bali.
Guntoro, S., I. M. Rai Yasa, I. N. Sumawa, Sumartini &
Rubiyo. 2002. Laporan Hasil Pengkajian Sistem
Usaha Tani Ternak Kambing dengan Tanaman
Industri. Proyek Pengkajian Partisipatif Bali. Balai
Pengkajian Teknologi Pertanian. Bali.
Laconi, E. B. 1998. Peningkatan Mutu Pod Kakao melalui
Ammoniasi dengan Urea dan Biofermentasi dengan
Phanerochaete chrysosporium serta Perjalanannya
ke dalam Formulasi Ransum Ruminansia. Thesis.
Program Pascasarjana. IPB. Bogor.
Mastika, I. M. 2006. Pengolahan Limbah Kakao sebagai
Pakan Alternatif dalam Laporan Akhir Demoplot
Pengendalian PBK dengan Pola Integrasi. DISBUN
dan HPT FAPERTA UNUD.
Parakkasi, A. 1999. Ilmu Nutrisi dan Makanan Ternak
Ruminan. Cetakan Pertama. Penerbit Universitas
Indonesia (UI-Press), Jakarta.
Preston, T.R. 1986. Better Utilization of Crop Residues and
By-product in Animal Feeding; Research Guideline
2. A Practical Manual for Research Worker; FAO
Animal Production and Health . Paper 50/2. FAO of
14 January 2011
GROWTH PERFORMANCE
United . Rome.
Purnama, I. N. 2004. Kajian Potensi Isolat Kapang Pemecah
Ikatan Tannin pada Kulit Buah Kakao (Theobroma
cacao L). Abstrak. Skripsi. Program Studi Nutrisi
dan Makanan Ternak. Departemen Ilmu Nutrisi dan
Makanan Ternak. Fakultas Peternakan. IPB. http://
fapet. ipb. ac. id [23 Mei 2007].
Suharto. M. 2004. Dukungan Teknologi Pakan dalam
Usaha Sapi Potong Berbasis Sumber Daya Lokal.
Makalah disampaikan pada Lokakarya Nasional Sapi
Potong. C.V. Lembah Hijau Multifarm, Surakarta.
http://peternakan.Litbang. deptan.go.id. [2 November
2006]
Sukanten, I. W., S. Putra & A. W. Puger. 1990. Pengaruh
Sistem Tiga Strata terhadap Penampilan Sapi Bali
dengan Program Peternakan Inti Rakyat. Proceeding
Seminar Nasional Sapi Bali. 21-22 September 1990.
Denpasar, Bali.
Indon. J. Nutr. and Feed Sci., January 2011, pages 15-19 Vol. 2 No. 1
ISSN 1410-2390

The Use of Cellulolytic Microbes from Cattle Rumen Fluid to Improve In Vitro
Digestibility of Fermented Robusta Coffee Pulp (Coffea canephora sp.)

L. M. Yusiati, C. Hanim and F. Az Zahra

Nutritional Biochemistry Laboratory, Faculty of Animal Science, Gadjah Mada University
Jl. Fauna No. 3 Bulaksumur, Yogyakarta 55281 Indonesia
(Received 09-11-2009 ; Accepted 14-10-2010)

ABSTRACT

The research was conducted to examine in vitro digestibility of fermented robusta coffee
(Coffea canephora sp.) pulp using rumen cellulolytic microbes. Cellulolytic microbes as much
as 0%, 5%, and 10% was added each to 200 g of coffee pulp (dry matter basis). Fermentation
was done anaerobically for 21 days at room temperature. The fermented coffee pulp was
examined for it’s odor, color, texture, pH, and the presence of fungi. The fermented robusta
coffee pulp samples were taken out and dried at 55oC, for analysis of dry matter (DM),
organic matter (OM), crude fiber (CF), crude protein (CP), ether extract (EE), and nitrogen
free extract (NFE), dry matter digestibility (DMD), organic matter digestibility (OMD), and
crude fiber digestibility (CFD). one way analysis of variance of completely randomized
design and continued by Duncan’s new multiple range tests were used to examine the
differences between mean values. addition of cellulolytic microbes 5% and 10% decreased
the fungi growth. Addition of 5% cellulolytic microbes did not affect the pH value (5.81 vs.
5.91). The addition of 10% cellulolytic microbes decreased pH from 5.81 to 5.57. Addition
of cellullolytic microbes 5% and 10% decreased CF content (P<0.01) as much as 9.14% and
21.12% from 42.99% (from the result of addition of 0% cellulolytic microbes) to 39.06% and
33.91% respectively. The NFE content of coffee pulp increased (P<0,01) by 56.83%, when it
was fermented by addition of 10% rumen cellulolytic microbes, while the addition of 5%
cellulolytic microbes did not affect NFE content. The increasing of inoculum from 5% to
10%, significantly increased NFE content by 24.21%. Nevertheless addition of cellulolytic
microbes did not give significant effect on DM, OM, CP and EE content of fermented coffee
pulp. The CFD, OMD, and DMD of fermented coffee pulp without addition of cellulolytic
microbes were 30.25%, 30.30% and 36.83%. Digestibility of crude fiber, organic matter and
dry matter increased significantly by 9.52%, 11.65%, and 8.85% after being treated with
5% and increased by 13.85% , 24.49%, 14.23% after being treated with 10% respectively.
It could be concluded that addition of 10 % cellullolityc microbes in robusta coffee pulp
fermentation altered physical and chemical composition as well as in vitro digestibility.

Key words : Fermented coffee pulp, cellullolytic microbes, in vitro digestibility

Introduction competitive uses with people. Various studies have

The problems facing by animal feeding to
support animal production in the developing countries
amongst which are poor feed quality, high prices and
*
Coresspondence author
Nutritional Biochemistry Laboratory, Faculty of Animal Science
Gadjah Mada University, Jl. Fauna Number 3 Yogyakarta 55281
E-mail : yusiati@yahoo.com
been done concerning with grass substitution by
agro-industrial by products, in animal diets. Krishna
and Umiyasih (2006) stated that there were five
inconventional feed materials that had potency as
cattle feed, they were cassava stem (756,761.14 ton/
year), maize cob (106,796.25 ton/year), soy bean
straw (106,692.40 ton/year), cocoa pod (1,103.86 ton/
year), and coffee pulp (312.42 ton/year). By product
of coffee is very interesting to be evaluated plantation

January 2011 15
YUSIATI ET AL.
for feedstuff substitution. Bressani (1979) reported
that the dry weight of coffee berries are represented
by 29% coffee pulp, 12% coffee hulls, 55% coffee
beans, and approximately 4% mucilage. Coffee pulp
disposal has created enviromental problem. Therefore,
several attempts have been made to re-use the coffee
by-products for different purposes, including animal
feeding (Braham and Bressani, 1979), production of-
amylase under solid state fermentation (Murthy et al.
2009) and cultivation and production of Pleurotus sp
by solid state fermentation (Murthy and Manonmani,
2008).
Representative values of the proximate chemical
composition of fresh pulp were dry matter 23%, crude
fiber 14.59%, crude protein 9.01%, ether extract
2.10%, nitrogen free extract 67.81% while chemical
composition of pulp 2-3 days after being collected
from the berry had dry matter 92%, crude fiber 22.61%,
crude protein 11.62%, ether extract 2.82%, nitrogen
free extract 53.47% (Elias in Braham and Bressani,
1979). Those proximate compositions show a relative
low nutritive value due to high level of crude fibre.
The fibrous parts of plants are made up exclusively
from cellulose, a polymer of D-glucose with β-
(14)- linkage. A variety of other polysaccharides
are present in plant cell walls. The cellulose fibrils are
cemented together by a matrix of three other polymeric
material, hemicelluloses, pectin and extensin (Mathew
and Holde, 1990). Fiber breakdown in the rumen is
a result of complex microbial processes involving
fibrolytic microorganisms includes bacteria, fungi and
ciliate protozoa. They produce a range of cellulolytic,
hemicellulolytic and pectinolytic enzymes which are
necessary for plant cell-wall degradation. Cellulose,
the main constituent of crude fibre, can be hydrolyzed
by cellulolytic microbes to its simplest constituent,
glucose. Major cellulolytic species in the rumen
fluid were Bacteroides succinogenes, Ruminococcus
flavefaciens, Ruminococcus albus and Butyrivibrio
fibrisolvens (Ishler, 1996).
One of the most significant factors which affect
digestibility is the chemical composition of feedstuff.
Therefore the changes in their chemical composition
as an effect of preservation, storage, maturity crop or
any others treatments will affect digestibility value. It
is generally believed that the percentage of crude fiber
affect nutrient digestibility. The greater the amount
of fiber, the lower the digestibility of the nutrient.
Application of fibrolytic inoculums had been used to
improve the quality of fibrous materials, such as rice
straw fermented by lignocellulolytic microbes (Yusiati
et al.,1995), palm cernel cake (Yusiati et al., 2009) and
caccao pod (Hanim et al., 2009) fermented by rumen
16 January 2011
Indon. J. Nutr. and Feed Sci.
cellulolytic inoculum.
Based on the above considerations, this study
was conducted to evaluate the effect of using rumen
cellulolytic microbes to ferment coffee pulp and
improve quality and in vitro digestibility of fermented
coffee pulp.
Materials and Methods
Microbe enrichment
Rumen fluid as a source of microbes was taken
out from fistulated rumen of Ongole Cross bred cattle
before morning feeding. Those fluid was examined
for carboxymethyl cellulase (CMC-ase) activity just
before used. The activity was measured based on the
amount of reducing sugar produced from the hydrolytic
reaction of CMC as a substrate by cellulolytic enzyme,
buffered at pH 5.5 The determination of reducing sugar
was carried out by ferricyanide reaction (Halliwel et
al., 1985). The CMC-ase activity was presented per
gram protein of the enzyme, which was measured by
Lowry’s Folin–Ciocalteu method using Bovine Serum
Albumin (BSA) as a standard (Plummer, 1971).
Specific activity was defined as activity per unit of
protein (unit/mg).
Rumen fluid as much as 10% was added into
medium suggested by Skinner (1971) that consisted of
0.2 g (NH4)2SO4; 0.1 g MgSO4.7H2O; 0.2 g K2HPO4;
0.4 g CaCO3; 2 ml NaCl 1 %; 1 g Cystein HCl; 2 drop
of resazurin (0.1%); 120 ml H2O; and 80 ml sterilized
rumen fluid and 2 g of cellulose. All steps during
preparation were carried out anaerobcally by flowing
CO2 gas into the fermenter. Fermentation was done at
39°C for 7 days.
Rumen cellulolytic microbes cultivation
The inoculum growing in the enrichment media
was taken out, and it was added into the glass fermenter
filled with growing medium as much as 10% from
the total medium. The medium consisted of 0.2 g
(NH4)2SO4; 0.02 g MgSO4.7H2O; 0.4 mg NaCl; 1.8339
g, K2HPO4.3H2O; 0.2 g yeast extract; 1 g cystein HCl;
2 drop of resazurin, 80 ml steril rumen fluid and 120
ml H2O. As much as 4 g of cellulose was added to the
medium as substrate (Skinner (1971). The process was
done totally in anaerobic condition by CO2 gassing.
The fermenters was kept at 39°C for 7 days. The
cellulolytic inoculum then was ready for coffee pulp
fermentation.
Vol. 2 No. 1
Fermentation of coffee pulp
Coffee pulp weighed 600 g were divided into
three parts. Rumen cellulolytic microbes as much as
0%, 5%, and 10% were added to each part of coffee
pulp (dry matter basis) respectivelly in 250 ml glass
silos. Fermentation was done anaerobically at room
temperature for 21 days. The fermentation was done
in three replicates.
Evaluation of fermented coffee pulp quality
At the end of the fermentation period the glass-
silos were opened for sampling. The samples were
examined for their odor, color, texture, pH, and the
presence of fungi. Content of dry matter was directly
determined from the fresh fermented samples prior to
55°C drying for measurements of nutrient digestibilty.
Fermented coffee pulp samples from each silo were
kept at −20°C for further analyses of organic matter,
crude fiber, crude protein and ether extract (AOAC,
1990) and also in vitro nutrient digestibility.
in vitro digestibility of nutrients determination
In vitro digestibility of nutrients in the fermented
coffee pulp, were evaluated in duplicate for each
sampel. The procedure included 48 h of incubation
of 0.5 g plant material with rumen fluid according to
the first step of two-stage fermentation technique of
Tilley and Terry (1963). Rumen fluid free of saliva
contamination was obtained before morning feeding
via rumen fistula from two cattle fed with King grass
and concentrate (70:30). Samples of the fermented
coffee pulp and the residues of digestion were assayed
in duplicate for content of DM (drying at 105°C for
24 h) and residual ash (4 h at 600°C) and crude fiber.
Based on the data obtained from the above analysis, in
vitro digestibility of dry matter (DMD), organic matter
(DMD), and crude fiber (CFD) could be calculated.
The data were subjected to statistical analysis by
adopting One Way Completely Randomized design,
and continued by Duncan’s New Multiple Range Test
(DMRT) (Astuti, 1981).
RESULTS AND DISCUSSIONS
Physical quality and pH of fermented Coffee Pulp
were shown in Table 1. The addition of cellulolytic
microbes 5% and 10% acid, it’s texture reduced, and
fungi growth but the brown color remained the same.
Addition of 5% cellulolytic microbes did not affect
the pH value (5.81 vs. 5.91), but the addition of 10%
cellulolytic microbes decreased pH from 5.81 to 5.57
(P<0.05).
THE USE OF CELLULOLYTIC MICROBES
Reduction of pH value might be due to increased
the lactic acid produced by lactic acid bacteria,
which were naturally found in the forages crop (Lynn
and Harrison, 2001). In homofermentative lactic
acid bacteria metabolism, the majority of glucose
present in the medium is converted to pyruvate via
glycolysis pathway, which is then reduced to form
lactic acid. The acid also produced by another group,
heterofermentative lactic acid bacteria from hexoses
or pentoses via pentose phosphate pathway, along
with volatile fatty acid, CO2, and ethanol. The acids
produced reduced the pH of medium.
The composition of fermented coffee pulp are
shown in Table 2 which demonstrated that addition
of 5% and 10% rumen cellulolytic microbes in coffee
pulp fermentation, decreased the crude fiber content
about 9.14% and 21.12% . The decreasing of crude
fiber content could be understood as the cellulolytic
microbes have abilities in degrading cellulose, a major
component of crude fiber. Cellulolytic activity of those
microbes was presented by CMC ase activity with the
value of 0.25 U/mg protein. Degradation of cellulose
became its monomer, glucose, was also be indicated
by increasing of nitrogen free extract (NFE) content
of fermented coffee pulp. The NFE content of coffee
pulp increased by 56.83%, when its fermentation was
using 10% of rumen cellulolytic microbes (P<0.01),
while the addition of 5% cellulolytic microbes did
not affect NFE content. The increasing of inoculum
from 5% to 10%, sinificantly increased NFE content
by 24.21%. The values of organic matter, CP and
EE were not affected. The decreasing values of CF
content as an effect of cellulolytic microbes, was in
agreement with Rojas et al. (2003) who reported that
cellulose content of ensilage Coffee pulp was lower
than oven dried-coffee pulp. Reduction of CF content
as an effect of fermentation also reported by Penaloza,
et al. (1985). It was reported that fermented Coffee
pulp had a higher total amino acid content and a lower
cell wall constituent value, primarily cellulose and
hemicellulose than the original pulp, after subjected to
a solid-state fermentation process.
Table 3 shows the nutrient digestibility of coffee
pulp treated with rumen cellulolytic microbes. The
nutrients digestibility including digestibility of
crude fiber, organic matter and dry matter increased
significantly by 9.52%, 11.65%, and 8.85% after being
treated with 5% and increased by 13.85%, 24.49%,
14.23% after being treated with 10 % of innocula
respectively. The increasing of CF digestibility was
considered due to the decreasing of CF content of
coffee pulp by fermentation. Luiza (2000) described
that cellulase is a complex mixture of enzyme proteins
January 2011 17
YUSIATI ET AL. Indon. J. Nutr. and Feed Sci.

Table 1. Physical quality and pH of coffee pulp after fermentation using rumen cellulolytic microbes

Addition of rumen cellulolytic microbes

Physical Quality
0% 5% 10%
Acidity Slightly acid acid acid
Color Brown Brown Brown
Texture Rough Smooth Smooth
Presence of fungi little Very little Very little
pH 5.91a 5.81a 5.57b
Note : Means in the same row with different superscript differ significantly (P <0.05)

Table 2. Chemical composition of coffee pulp after fermentation using rumen cellulolytic microbes (% DM)

Chemical Composition Addition of rumen cellulolytic microbes

Significant
(%) 0% 5% 10%
Dry Matter 59.74 59.70 63.19 ns
Ash 12.76 13.39 12.99 ns
Crude Fiber 42.99 C
39.06 B
33.91 A
P <0.01
Crude Protein 29.05 28.63 27.31 ns
Ether Extract 2.65 3.24 3.00 ns
Nitrogen Free Extract 16.70 a
19.85 a
26.19 b
P <0.05

Table 3. Nutrient digestibility of coffee pulp after fermentation (%)

Digestibility Addition of rumen cellulolytic microbes Significant

0% 5% 10%
Crude fiber 30.25A 33.13B 34.44C P <0.01
Organic matter 30.30A 33.83B 37.72C P <0.01
Dry matter 36.83a 40.09b 42.07b P <0.05

with different specificities to hydrolyze glycosidic
bonds. The three major cellulase enzyme are:
endocellulase (1,4- ß-D-glucan- 4-glucanohydrolase,
endoglucanase, EG, EC 3.2.1.4.), exocellulase (1,4-
β-D-glucan-cellobiohydrolase, CBH, EC 3.2.1.91)
and β-glucosidase (β-D-glucosido-glucohydrolase,
cellobiase, EC 3.2.1.21). Those enzymes break the
linkages between components of crude fiber, therefore
other enzymes produced by rumen microbes are
facilitated to digest any compounds inside the cell. As
the result, the digestibility of organic and dry matter
increased.
Conclusion
The use of rumen cellulolytic microbes at the
level of 5% and 10% in coffee pulp fermentation
increased physical and chemical quality as well as in
vitro digestibility. The addition of rumen cellulolytic
microbes at the level of 10%, resulted the highest
qualities and digestibility of coffee pulp.
REFERENCES
Association of Official Analytical Chemists (AOAC). 1990.
Official Methods of Analysis of the Association of
Official Analytical Chemists, Food Composition;
Additives; Natural Contaminants. Helrich, K. (Ed.),
15th Edition. Virginia, USA, AOAC, Inc.
Astuti, M. 1981. Rancangan Percobaan dan Analisis
Statistik. Bagian I. Fakultas Peternakan. Universitas
Gadjah Mada. Yogyakarta.
Braham, J. E. & R. Bressani. 1979. Coffee Pulp :
Composition, Technology, and Utilization. Institute
of Nutrition of Central America and Panama.
Bressani, R. 1979. The by-products of coffee berries.
In : Coffee Pulp : Composition, Technology, and

18 January 2011
Vol. 2 No. 1
Utilization. J.E. Braham and R. Bressani. (Eds).
Institute of Nutrition of Central America and Panama
Elias. 1979, Chemical composition of coffee-berry by-
products. In:Coffee Pulp: Composition, Technology,
and Utilization. J.E. Braham and R. Bressani.(eds)
Institute of Nutrition of Central America and Panama
Halliwel, G., N. N. B. A. Wahab & A. H. Patle. 1985.
Chemical Compotition of endo 1,4-β-glucanase to
cellulolitic in trichodrma koningii. J. App. Biochem.
7: 43-45.
Hanim. C., L. M. Yusiati, & V. P. Budyastuti. 2008. Pengaruh
Penambahan Mikrobia Selulolitik dari Cairan Rumen
Sapi pada Fermentasi Kulit Buah Kakao (Theobroma
cacao) terhadap Kecernaan in vitro dengan Metode
Produksi Gas. In Progress.
Ishler, V. 1996. From Feed To Milk: Understanding Rumen
Function. College of Agricultural Sciences The
Pennsylvania State University. Pp.5-6.
Krishna, N.H. & U. Umiyasih .2006. Identification and
Evaluation Nutrient of Non-Conventional Feed
Material from Abundant Waste in Daerah Istimewa
Yogyakarta District. Seminar Nasional Teknologi
Peternakan dan Veteriner PP.872-879.
Lynn M. J.& J. H. Harrison. 2001. Scientific Aspects of
Silage Making. Proceedings, 31st California Alfalfa &
Forage Symposium, Modesto, CA, UC Cooperative
Extension, University of California, Davis.
Luiza, J.2000. Solid state fermentation of agricultural wastes
for endoglucanase production. Int. J. of Industrial
Crops and Products 11 :1–5.
Mathew, C.K. & K.E. Van Holde, 1990. Biochemistry.
The Benjamin/ Cummings Publishing Company 390
Bridge Parkway. Redwood City, CA 94065.
Murthy P.S. & H. K. Manonmani. 2008. Bioconversion
of Coffee Industry waste with White Rot Fungus
THE USE OF CELLULOLYTIC MICROBES
Pleurotus florida. Research J. Environ. Sci. 2(2): 145-
150
Murthy, P. S., M. M. Naidu & P. Srinivas. 2009. Production
of -amylase under solid-state fermentation utilizing
coffee waste. J. Chem.Technol. Biotechnol. 84 (8):
1246 – 1249
Penaloza, P., M. R. Molina, R. G. Brenes & R. Bressani.
1985. Solid-State Fermentation: an Alternative to
Improve the Nutritive Value of Coffee Pulp. J. Appl
and Environ. Microbiol. 49 (2): 388-393
Plummer, D. T. 1971. An Introduction to Practical
Biochemistry. Tata Mc Graw-Hill Publishing
Company Ltd., Bombay-New Delhi
Rojas, J. B. O., J. A. J. Verreth, S. Amato & E.A. Huisman.
2003. Biological treatments affect the chemical
composition of coffee pulp. Biores. Technol. 89: 267–
274
Skinner, F. A. 1971. Isolation of Soil Clostridia. In: Isolation
Of Anaerobes. The Society for Applied Bacteriology
Technical Series no 5. Academic Press, London, New
York.
Tilley, J. M. & R. M. Terry. 1963. A two stage technique
for the in vitro digestion of forage crops. J. Br. Grass.
Soc.18, 104–111.
Yusiati, L.M., Z. Bachrudin, Kustono & Didi Rachmadi.
1995. Chemical evaluation of lignocellulolitic
microbes, yeast,and lactobacili addition to rice straw
at silage preservation. Bulletin of Animal Science.
Special Edition, ISSN 0126-4400.
Yusiati, L. M., Wihandoyo & C. Hanim, , 2009.
Pemanfaatan Bungkil Kelapa Sawit Fermentasi
Menggunakan Isolat Fibrolitik dalam Pakan Itik
Terhadap Produksi dan Kualitas Telur. In progress.
January 2011 19
Indon. J. Nutr. and Feed Sci., January 2011, pages 20-25 Vol. 2 No. 1
ISSN 1410-2390

Improving the Nutritive Value of Total Mixed Ration Based on By-products

Fermented by Rumen Liquor and Enzyme

I. M. Mudita, A. A. P. P. Wibawa, I. W. Wirawan, N. W. Siti and I. G. L. O. Cakra

Faculty of Animal Husbandry, University of Udayana
Jl. PB Sudirman Denpasar Kampus Bukit Jimbaran 80361, Bali-Indonesia
(Received 09-11-2009 ; Accepted 24-08-2010)

ABSTRACT

An experiment was carried out to optimize the nutritive value of total mixed ration based
on by-products fermented with rumen liquor and enzyme complex. Factorial Completely
Randomized Design was used through out the experiment. The level of rumen liquor of 0
ml (RL0), 20 ml (RL20) or 40 ml (RL40) per 1 kg total mixed ration; the levels of optyzim as
sources of enzyme complex of 0g (E0), 1 g (E1) or 2 g (E2) per 1 kg total mixed ration . The
treatments were allocated randomly in 3x3 factorial of completely randomized design in
three replicates. Nutritive value of total mixed ration including contents of dry matter/DM,
organic matter/OM, crude protein/CP and crude fibre/CF were determined. Fermentation
products including concentration of NH3-N, total VFA, acetic acid, propionic acid, butyric
acid, and methane were also measured. There was treatments interaction (P<0.05) effect
on nutritive value except dry matter content, concentration of acetic and butyric acid.
The untreated total mixed ration (RL0), optimum indicated good nutritive value (P<0.05)
Fermentation of local by-products as compound of complete feeds using rumen liquor and
enzym complex reduced contents of organic matter, crude protein, crude fiber, pH, and
methane gas production, while concentrations of total VFA, propionic acid and NH3-N
increased.

Key words : Complete feed, enzyme complex, local waste, nutrition quality, rumen liquor

INTRODUCTION Most of agricultural by-products have limited

The development of feeding system based on the
local resources is the pillars supporting the sustainable
and competitive production systems especially animal
species in Indonesian (Ginting, 2004). By-products
of food crops and agro-industry are potential sources
of ruminant feed ingredients. Total mixed ration for
ruminnt can be formulated and compound of the by-
product available localy (Wahyono and Hardianto,
2007).
*
Coresspondence author
Faculty of Animal Husbandry, University of Udayana
Jl. PB Sudirman Denpasar Kampus Bukit Jimbaran 80361 Bali
E-mail : muditafapet_unud@yahoo.com
nutritive value (high fiber, low available nutrient
and digestibility), contain various anti-nutritional
compounds such as oxalic, silica, tannins, theobromine,
cyanide, and ceratyne which can lead to decrease
productivity of livestock (Preston, 1986), thus they
become a limiting factor in the utilization in rations
(Ginting, 2004). . Fermentation is the most method
strategy which can be applied to improve the nutritive
value of the by-products (Tamada et al., 1999).
Utilization of inoculants containing the degrading
fibrous microbes such as rumen liquor and the enzyme
complex in the fermentation may nutritive value of
total mixed ration compound of local by-products.
Rumen liquor is animal from slaughterhouse waste
containing a variety bacteria, protozoa and fungi (Arora,
1995) and produce different types of fiber degrading
enzymes, such as alpha-amylase, galactosidase,

20 January 2011
MUDITA ET AL.
hemicellulases, cellulase, and xylanase (Kamra, 2005).
Optyzim is a comercial enzyme complex in powder
form consisting of a mixture of amylase, protease,
xylanase, cellulase, and hemicellulases (Bidura, 2006).
Inclution of inoculants hydrolysis plant cell walls
(cellulose, hemicelluloses, xylan and pectin), reduced
Neutral Detergen Fiber (NDF) and Acid Detergen
Fiber (ADF) content (Parakkasi, 1999), increased
digestibility of crude protein, energy, dry matter, and
NDF of feed ingredients (Hau et al., 2006).
Pantaya et al. (2005) showed the addition of
enzymes from rumen liquor reduce the polysaccharide
content and increased metabolized energy content
of wheat pollard. Purnomohadi (2006) reported that
addition of celulolytic bacteria of the rumen liquor
improved crude protein (4.1% to 9.01%) and decreased
crude fiber (37.10% to 31.17%). Lamid et al. (2006)
reported that the use of xylanolytic bacteria (5-15%)
of rumen liquor in rice straw ensilage increased crude
protein and reduce a crude fiber content of rice straw
silage. Ridla and Uchida (1999) reported that the
combined treatments of lactic acid bacteria (LAB) and
celulase improved the fermentation quality of both
the Rhodesgrass and Italian Ryegrass. by degrading a
portion of cell wall during storage.
Nahrowi (2006) showed the assignment of silage
complete feed, was accepted by sheep, beef and dairy
cattle, with no negative impact on performance. Feeding
silage became the source of probiotics, defaunating
agents and alternative antibiotics for livestock which
affected the balance of rumen microbes (Hau et al.,
2006; Wina, 2006). Krisnan and Ginting (2005)
reported that inclusion of fermented markisa 20 to
40% in ration gave the same intake, feed efficiency
and daily gain of Kacang goat compared to control.
Parwati et al. (2006) showed that goat production
given feed ration containing fermented cashew waste
improved.
This experiment was conducted to determine the
best level of rumen liquor and enzyme complex in the
fermentation to improved nutritive value of total mixed
ration complete feed compound of by-products.
MATERIALS AND METHODS
Location, feed and experimental design
An experiment was carried out at Bukit
Jimbaran Research Stasiun and Animal Nutrition
Laboratory, Faculty of Animal Husbandry, Udayana
University. Experimental treatments had two factors
as follows: (1) RL0 = without rumen liquor, RL20 = 20
ml rumen liquor, and RL40 = 40 ml rumen liquor per 1
Indon. J. Nutr. and Feed Sci.
kg complete feed; (2) the levels of 0, 1 and 2 optizyme
as a source of enzyme complex per 1 kg complete
feed as E0, E1 or E2, respectively. Each combination
of treatment had three replicates. The rumen liquor
and/or enzyme optizyme were diluted in 1 liter fresh
clean water for each kg of a complete feed. Basal
complete feeds composed of local feedstuff (Table 1).
The complete feeds were treated with rumen liquor
and optizyme and incubated for 31 days at anaerobic
condition. Fermentability of complete feeds from
different treatments was evaluated in vitro. The
treatments were allocated in a Factorial Completely
Randomized Design of 3 x 3 with three replications.
Data collection, sampling procedure and analysis
Nutrient content of the complete feed including
dry matter (DM), organic matter (OM), crude protein
(CP) and crude fibre (CF) were determined. The
characteristics of the complete feed fermentability
were evaluated. Concentration of NH3-N, total VFA,
partial VFA (acetic acid, propionic acid, butyric acid),
pH. and methane gas production were determined and
used as variables.
Feed samples were collected from untreated
and treated complete feeds produced and used in the
experiment. Feed samples were analyzed for DM,
ash, CP and CF contents according to the proxymate
analysis method of AOAC (1980). Concentration
of NH3-N was analysed using the phenolhypoclorite
method, Total volatile fatty acids was analysed bythe
method of steam destilation technique (Department of
Dairy Science, 1966), pH was measured using portable
pH meter “HANNA instrument HI 9025”, partial VFA
(Acetic, propionic and butyric) was analysed using
a high performance liquid chromatography HPLC,
and methane production was estimated accoding to
the following calculaton: CH4 = 0.5 acetate – 0.25
propionate + 0.5 butirate production (Orskov dan
Ryle, 1990).
Data collected were analyzed statistically
by applaying the analysis of applied if necessary
(Sastrosupadi, 2000).
RESULTS AND DISCUSSIONS
Fermentation with rumen liquor or optyzime
addition in a complete feed decreased (P<0.05) the
contents of organic matter/OM, crude protein/CP and
crude fiber/CF. There was the effect of interaction
(P<0.05) between the level of rumen fluid and optyzime
addition on these variables, except on dry matter
(P>0.05). The interaction of the treatments (P<0.05)
January 2011 21
Vol. 2 No. 1 IMPROVING THE NUTRITIVE QUALITY

changed significantly ruminal pH, concentration of acid.

NH3-N, total VFA, propionic acid and methane. The
concentration of acetic acid and butyric acid were
not influenced (P>0.05) by the treatments and their
interaction (Table 2-4). The results indicated that
inoculants and optyzime had complementary effect
in improving the nutritive value of the complete feed
when they were added in it. This was probably also
due to the increasing production of total propionic
Addition of both rumen liquor and optizyme
decreased OM, CP, and CF content, and reduced acidity
and methane production in vitro, but increased NH3-
N, total VFA production and propionic acid proportion
in the VFA (Table 4). Reduction of OM, CP, and CF
content in the complete feed was likely associated
with the increased in microbial activity. The microbes
in rumen liquor added, fermented and utilized the

Table 2. Nutritive Value of Complete Feeds Treated with Inoculants and Optizyme
Treatment Nutritive Value
DM OM CP CF
---------------------------------------------------------- % DM ------------------------------------------------------------
Rumen Liquor
RL0 96.49a 81.90b 17.62c 23.84c
RL20 96.19a 80.19a 16.44a 23.21b
RL40 96.52a 80.92a 16.75b 22.90a
SEM 0.45 0.21 0.04 0.06
Optyzime
E0 96.79a 81.80b 17.40c 24.34c
E1 95.72a 80.61a 16.87b 23.22b
E2 96.69a 80.60a 16.55a 22.38a
SEM 0.45 0.21 0.04 0.06
Interaction P>0.05 P<0.05 P<0.05 P<0.05
Note: The means in same column with different letter different (P<0.05); SEM = Standard Error of the treatment means.

Table 3. Fermentation Characteristics of Complete Feeds treated with Inoculants and Optyzim

Fermentation Characteristic
pH NH3-N Total VFA Acetic Propionic Butyric acid Methane
Treatment acid acid
------------------------------------------------------- mM-----------------------------------------------
Rumen Liquor

RL0 7.31c 11.88a 126.52a 55.88a 28.70a 11.12a 26.33a

RL20 6.62a 13.13b 179.15b 54.48a 30.39b 10.77a 25.03a

RL40 6.81b 12.54ab 176.11b 54.86a 30.93b 10.07a 24.73a

SEM2 0.01 0.26 2.55 0.72 0.40 0.71 0.44
Optyzime 

E0 7.38c 11.97a 141.58a 55.66a 29.16a 10.45a 25.77a

E1 6.88b 12.55ab 164.81b 54.27a 30.91b 10.23a 24.52a

E2 6.47a 13.03b 175.39c 55.29a 29.95ab 11.29a 25.80a

SEM 0.01 0.26 2.55 0.72 0.40 0.71 0.44
Interaction P<0.05 P<0.05 P<0.05 P>0.05 P<0.05 P>0.05 P<0.05
Note: The means in same column with different letter different (P<0.05); SEM = Standard Error of the treatment means.

22 January 2011
MUDITA ET AL. Indon. J. Nutr. and Feed Sci.

nutrients composing the complete feed. The decreased
of OM and CP of treated complete feeds could be due
to the utilization of nutrients in the feed by microbial
inoculants for microbial protein synthesis to grow and
support its activities (Fellner, 2004; Leng, 1997).
Increased NH3-N, total VFA production and its
propionic acid proportion indicated that addition
of inoculant and optizyme into the complete feed
increased the fermentability of the feed. Activity
of microbe inoculated into the feed hydrolyzed the
large molecules of nutrients of the complete feed.
The decreased in fiber content of treated feed up
to 2 g optizym in 1 kg complete feed and high pH
liquor in the in vitro test, indicated the effectiveness
of the enzyme produced by inoculant and optizyme
in hydrolyzing the feed components including fiber
compenents. The decreased in rumen pH liquor
indicated that the inoculant and optizyme increased
the solubility and availability of fiber, protein and
others feed components, and therefore increased fiber
fermentation and the protein degradation in the rumen
liquor. The high acidity of pH 5.81 had been providing
the best conditions for cellulolytic microbial activity
to produce the highest total VFA (Fellner, 2004).

Table 4. Nutritive Value of Complete Feeds Treated with Inoculants and Optizyme

Parameters Rumen Liquor Optyzime SEM

Observed
E0 E1 E2
OM RL0 84.37Bb 80.95Aa 80.37Aa 0.36
(% DM basis) RL20 80.28 Aa
80.19 Aa
80.10 Aa

RL40 80.74Aa 80.71Aa 81.32Aa

CP RL0 18.82Cc 16.54Aa 17.51Bc 0.08
(% DM basis) RL20 17.17Cb 16.44Ba 15.71Aa
RL40 16.20Aa 17.62Bb 16.44Ab
CF RL0 25.96Cc 23.33Ba 22.22Aa 0.11
(% DM basis) RL20 23.79 Cb
23.21 Ba
22.63 Ab

RL40 23.26Ba 23.13Ba 22.30Aab

Acidity RL0 7.58Cc 7.30Bc 7.04Ac 0.01
(pH)
RL20 7.42Cb 6.62Ba 5.81Aa
RL40 7.14Ca 6.73Bb 6.57Ab
NH3-N RL0 10.65Aa 11.51Aa 13.47Ba 0.45
(mM)
RL20 13.10 Ab
13.13 Aa
13.17 Aa

RL40 12.17Aab 13.00Aa 12.45Aa

Total VFA RL0 84.74Aa 138.80Ba 156.02Ca 4.41
(mM)
RL20 171.08Ab 179.15ABb 187.22Bb
RL40 168.93Ab 176.46Ab 182.92Ab
Propionic Acid RL0 25.14Aa 30.62Ba 30.34Ba 0.69
(mM)
RL20 31.59Ab 30.03Aa 29.55Aa
RL40 30.74Ab 32.08Aa 29.97Aa
Methane RL0 28.44Bb 25.13Aa 25.41Aa 0.77
(mM)
RL20 23.96Aa 25.51Aa 25.61Aa
RL40 24.90ABa 22.92Aa 26.37Ba
Note : The means in the same row or column with different capital or small letter respectively differ (P<0.05). SEM = Standard Error of
the Treatment Means

January 2011 23
Vol. 2 No. 1
The increased in propionic acid proportion in the
VFA was likely associated with the reduction of crude
fiber components in the treated complete feed. The
decline in fiber will generally be accompanied by an
increase in feed digestion (Fellner, 2004; Leng, 1997).
Increased in the fermentability of the feed indicated
that the inoculation of microbes and addition of
optizyme improved the nutritive value of the complete
feed (Mudita, 2008). The improvement of nutrients
availability in the complete feed produced a better
fermentation condition and facilitated the activities
of microbes releasing fiber degrading enzymes during
fermentation of the components of complete feed in
the rumen liquor (Bidura, 2006).
Methane is one of the major components of
environmental pollutant (Hegarty, 2001). The reduction
in methane production in the present experiment
indicated that the addition of inoculants and optizyme
into the complete feed contributed to reduction of
methane released from the ruminants. These conditions
have also given a very positive effect in which resulted
complete feed potentially produce the lowest methane
production so that it is feasible to be developed in the
effort of developing ruminant farm environmentally
friendly. The low production of methane will reduce
the risk of environmental pollution from ruminant
farm which is currently contributing high pollutants
that is 20-30% of the total emission of methane into
the atmosphere (Nurtjahya et al., 2006) or it reaches
62.6 Mt CO2-equivalent (Hegarty, 2001).
CONCLUSION
Microbe from rumen liquor utilized some
components of the complete feed for their growth
and activities, and therefore reduced the organic
matter, crude protein, and crude fiber content of feed.
However, the addition of rumen liquor in combination
with optizym were able to hydrolyze the feed
components and improved the availability as well as
the fementability of the complete feed components.
REFERENCES
Arora, S.P. 1995. Pencernaan Mikroba Pada Ruminansia.
Translated by: Retno Murwani: Microbial Digestion
in Ruminants. Gadjah Mada University Press,
Yogyakarta
Association of Official Analytical Chemist (A.O.A.C.). 1980.
Official Method of Analysis. 13th Ed., Washington,
DC.
24 January 2011
IMPROVING THE NUTRITIVE QUALITY
Bidura, I.G.N.G.. 2006. Feed Animal Biotechnology.
Subjects for Tuition. Faculty of Animal Husbandry,
Udayana University, Denpasar (In Indonesian)
Department of Dairy Science, 1966. General Laboratory
Procedures. University of Wisconsin, Madison.
Fellner, V.. 2004. Rumen Microbes and Nutrient
Management. Animal Science Departmental Report.
North Carolina State University. http://mark.asci.ncsu.
edu/SwineReports/2004-2005/DairyCattle/Nutrition/
Fellner1.html [December 20, 2007].
Ginting, S.P.. 2004. Challenges and Opportunity in The
Utilization of Local Feeds for Goat Production in
Indonesian. Article in Lokakarya Nasional Kambing
Potong. http://peternakan.litbang.deptan.go.id/down-
load/infoteknis/kambingpotong/prokpo04-7.pdf.
[January 30, 2007]. (in Indonesian with abstract in
English)
Hau, D.K., M. Nenobais, J.Nulik, N.G.F. Katifana.. 2006.
Effect Probiotic of Digestibility Rumen Microbes of
the Bali Cattle. http://peternakan.litbang.deptan.go.id
[December 24, 2007]. (in Indonesian with abstract in
English)
Hegarty, R.. 2001. Green House Gas Emissions From The
Australian Livestock Sector. What Do We Know,
What Can We Do?. Australian Greenhouse Office.
ISBN: 1 876536 69 1. Canberra.
Kamra, D. N. 2005. Rumen Microbial Ecosystem. Special
Section: Microbial Diversity. Current Science
89 (1). Pp:124-135. http://www.ias.ac.in/currsci/
jul102005/124.pdf [December 20, 200].
Krisnan, R., and S. P. Ginting. 2005. The Productivity of
Kacang Goat Using Aspergillus niger Fermented
Markisa Rind (Passiflora edulis Sims. Edulis Deg.)
as a Complete Feed. Article: in Seminar Nasional
Teknologi Peternakan dan Veteriner. Pp:625-629. (in
Indonesian with abstract in English).
Lamid, M., S. Chuzaemi, N.N.T. Puspaningsih and
Kusmartono. 2006. Innoculation of Rumen’s
Xyalolytic Bacteria to Improve Rice Straw Nutritive
Value. In: Journal Protein 14 (2).Pp:122-128. Http://
jurnal-protein.umm.ac.id [January 2, 2010]. (in
Indonesian with abstract in English).
Leng, R. A.. 1997. Tree Foliage In Ruminant Nutrition. Food
and Agriculture Organization of The United Nations
Rome, Italy. http://www.fao.org/docrep/003/w7448e/
W7448E00.html [September 14, 2007].
Mudita, M. 2008. Rumen Microbial Protein Synthesis Of Bali
Cattle Given Complite Ration Based on Ammoniated
Rice Straw with Supplementation of Multivitamins-
Minerals. Thesis. Graduate School (Magister),
Udayana University, Denpasar. (in Indonesian with
abstract in English).
Nahrowi. 2006. Silase Ransum Komplit: Strategi Penyediaan
Pakan Ternak Ruminansia Berkelanjutan. Materi
Pelatihan. IPB, Bogor.
MUDITA ET AL.
Nurtjahya, E. S. D.Rumetor, J. F. Salamena, E. Hernawan,
S. Darwati, S. M. Soenarno. 2006. Utilization Animal
Waste for Reduced Environment Pollution. Article for
Lesson ”Pengantar Falsafah Sains”. Doctoral School,
IPB. http://tumoutou.net/6_sem2_023/kel4_seml_02
[December 16, 2006]. (in Indonesian)
Orskov, E.R. and M. Ryle. 1990. Energy Nutrition in
Ruminants. Elsevier Applied Science. London.
Pantaya, D., Nahrowi, L. A. Sofyan. 2005. Suplementation
of Enzyme Rumen Liquor on Feed Wheat Pollard
Based Diet on Processing Feed with Steam Pelleting
on Feed Chemical Composition. In: Proceeding
of National Seminar on Indonesia Assosiation of
Nutrition and Feed Science V, Brawijaya University,
Malang. (in Indonesian with abstract in English)
Parakkasi, A.. 1999. Ilmu Nutrisi dan Makanan Ternak
Ruminan. Penerbit Universitas Indonesia, Jakarta. (in
Indonesia)
Parwati, I. A., S. Guntoro, R. Yasa and I N. Suyasa. 2006.
The Impact of Fermented Chashew Waste on the
Farmer Income in Juntal Village Kubu Karangasem.
Article: in Seminar Nasional Teknologi Peternakan
dan Veteriner. Pp:561-566
Preston, T.R. 1986. Better Utilization of crop Residues and
By-product in Animal Feeding; Research Guideline
2. A Practical Manual for Research Worker; FAO
Animal Production and Health . Paper 50/2. FAO of
United . Rome
Purnomohadi, M. 2006. The Role of Rumen Cellulolytic
Bacterial Inoculants on The Fermentation Process of
Feed Quality. In: Journal Protein 13 (2).Pp:108-112.
http://jurnal-protein.umm.ac.id [ January 2, 2010]. (in
Indonesian with abstract in English).
Indon. J. Nutr. and Feed Sci.
Ridla, M. and S. Uchida. 1999. Comparative Study on
The Effects of Combined Treatments of Lactic
Acid Bacteria and Celases on The Fermentation
Characteristic and Camical Composition of
Rhodesgrass (Chloris gayana Kunth.) and Italian
Ryegrass (Lolium multiflorum Lam.) Silages. Asian-
Australasian J. Anim. Sci 12 (4). Pp:525-530.
Sastrosupadi, A.. 2000. Rancangan Percobaan Praktis
Bidang pertanian. Edisi Revisi. Penerbit Kanisius,
Yogyakarta. (in Indonesian)
Tamada, J., H. Yokota, M. Ohshima and M. Tamaki. 1999.
Effect of Additives, Storage Temperature and Regional
Difference of Ensilasing on fermentation Quality of
Napier Grass (Pennisetum purpureum Schum.) Silage.
Asian-Australasian J. Anim. Sci. 12 (1):28-35.
Wahyono, D. E., and R. Hardianto. 2004. Utilization of
Local Feed Resources to Develop Beef Cattle. Article
in: Lokakarya Nasional Kambing Potong. Http://pe-
ternakan.litbang.deptan.go.id/download/infoteknis/
kambingpotong [January 30, 2007]. (in Indonesian
with abstract in English)
Wina, E. 2005. The Technology of Utilizing Microorganism
in Feed to Improve Ruminant Productivity In
Indonesia: A Review. http://peternakan.litbang.deptan.
go.id/publikasi/wartazoa/wazo154-2.pdf [December
24, 2006]. (in Indonesian with abstract in English)
January 2011 25
Indon. J. Nutr. and Feed Sci., January 2011, pages 26-31 Vol. 2 No. 1
ISSN 1410-2390

The evaluation of nutrient quality of ramie leaves silage and hay in complete mixed
ration for Etawah-Crossbreed goat using in vitro technique

Despal*, I. M. L. Hutabarat., R. Mutia, and I. G. Permana

Department of Nutrition and Feed Technology
Faculty of Animal Science, Bogor Agricultural University, Bogor
(Received 09-11-2009. ; Accepted 23-03-2010)

Abstract
A research was conducted to evaluate the effect of ramie leaves silage and hay in Etawah
Crossbreed (PE) goat complete mixed ration (CMR) on nutrient content, fermentability, and
digestibility by in vitro. There were seven CMR dietary treatments i.e. P0 (control ration)
= 50% napier grass + 50% concentrate, P1 = 30% napier grass + 20% ramie leaves silage +
50% concentrate, P2 = 20% napier grass + 30% ramie leaves silage + 50% concentrate, P3
= 10% napier grass + 40% ramie leaves silage + 50% concentrate, P4 = 30% napier grass +
20% ramie leaves hay + 50% concentrate, P5 = 20% napier grass + 30% ramie leaves hay
+ 50% concentrate, and P6 = 10% napier grass + 40% hay + 50% concentrate. Both ramie
leaves silage and hay increased the CMR digestibility and nutrient content, except the crude
fiber. Control ration had a higher crude fiber than silage and hay. The CMR which contain
40% ramie leaves silage had higher nutrient digestibilities compared to the other rations.
Rations which were added with ramie leaves silage (P1 – P3) had higher VFA concentration
compared to the other rations. Ammonia concentration of rations added with preserved
ramie leaves were lower than control, however ammonia concentration in all treatments
were in optimal range. Acetate proportion was higher in CMR which contain ramie leaves
hay than CMR which contain silage and the nutrients digestibilities were lower. Adding
ramie leaves silage in rations resulted higher propionate and butyrate proportion than
control and rations added with ramie leaves hay. Either silage or hay ramie leaves can be
used up to 40% as napier grass substitute in the PE CMR.

Key words : Etawah goat, hay, ramie leaves, silage

Introduction animal (Duarte et al., 1997). Sufficiently high crude

Ramie leaves are byproduct from ramie
(Boehmeria nivea) plantation that produced fiber for
textile raw materials. Currently, ramie plantations are
widely expanded in Garut and Wonosobo regencies.
The previous research showed that ramie leaves
contained all major nutrients which were needed by
*
Coresspondence author
Department of Nutrition and Feed Technology
Faculty of Animal Science, Bogor Agricultural University
E-mail : despal04@yahoo.com
protein content (20%) and crude fibre (16%) exhibited
that ramie leaves could be used as forage to fulfill
dairy nutrient requirement like PE goat. Despal (2007)
explained that supplementation of dried ramie leaves
until 33% in ration based on field grass prevented
sheep body weight loss during dry season and gave
positive growth.
Ramie leaves available periodically depend on
stem harvest at 25 – 40 days interval. Harvesting occur
at the same time and in great quantity. Each hectare
of ramie plantation could produce forages up to 300
ton fresh material/year (FAO, 2005) or equivalent to
42 ton dry matter. Preservation of ramie leaves was

26 January 2011
DESPAL ET AL.
necessary so that ramie leaves could be utilized more
efficiently and being used as animal daily feed.
General preservations of forages are wet (silage)
and dry (hay) preservations. Each technique has
advantages and disadvantages. Drying with open sun
drying technique is a cheap forage conserving method.
However, forage excess generally occur at rainy
season so there is a needed for technology to handle
the constraint. Whereas wet preservation (silage) is
hampered by low water soluble carbohydrate (WSC)
and high water content that may produce a low quality
of silage.
According to Despal and Permana (2008), ramie
leaves dried by greenhouse technique produced better
quality of hay than drying by open sun drying and
oven technique. Adding dried cassava 20% (w/w) in
silage ramie leaves produced better quality of silage
than silage which were added with corn and pollard.
The quality of preserved ramie leaves needed to be
tested in ration.
The objective of the research was to study
preserved ramie leaves using wet and dry preservation
as grass substitute in PE goat ration and their effect
on nutrient content, fermentability and in vitro
digestibility.
Materials and Methods
This research was conducted from November
2008 to March 2009 at Agrostology Laboratory, Dairy
Animal Nutrition Laboratory, Department of Nutrition
and Feed Technology, Faculty of Animal Science,
Bogor Agricultural University, Laboratory of Inter
University Center, Bogor Agricultural University, and
Laboratory of Nutrition Physiology, Animal Research
Center, Ciawi.
Ramie leaves were obtained from Koperasi
Pondok Pesantren (Koppontren) Darusalam, Garut
Regency. As many as 2 kg of ramie leaves, that was
chopped into a length of approximately 1,5 – 2 cm
using forage chopper, added with 400 grams of dried
cassava to make the silage. Silage was stored in plastic
(28 x 50 cm) and rewrapped with plastic and polybag
(60 x 120 cm) to avoid light intervention. Silages were
incubated anaerobically for 35 days. After 35 days,
silages were dried, ground, and mixed in ration. Hay
was made by drying ramie leaves in greenhouse for
21 hours under intensive light and the hay was twist
every 2 hours. After 21 hours light intensities, hay was
ground and mixed in ration. The forage which used
in ration was napier grass whereas the concentrate
consisted of corn, pollard, rice bran, pressed coconut
cake, dried cassava, CaCO3, and dicalsium phosphat.
Indon. J. Nutr. and Feed Sci.
Chemical composition of ingredients which were used
in complete mixed ration was appeared in Table 1.
Complete ration was mixed appropriately
according to formula (Table 2). Complete ration
was formulated based on the nutrient requirement of
lactating PE having 30 kg BW and produce 1 kg milk/d
(4% FCM). The ration contained 66.5% TDN, 11.17%
CP, 0.41% Ca, and 0.29% P (NRC, 1981).
Nutrients content, i.e. dry matter (DM), crude
protein (CP), crude fibre (CF), ether extract (EE),
and ash were analyzed according to AOAC (1999).
Fermentability and in vitro digestibility were
determined as described by Tilley and Terry (1969),
NH3 analysis was conducted according to General
Laboratory Procedure (1966), and partial VFA were
analyzed with gas chromatography using Chrompack
method (1998).
Data were subjected to analysis of variance
(ANOVA). Significant differences between individual
means were identified using Duncan’s multiple tests.
Results and Discussions
Nutrient composition of complete ration
Proximate composition is presented in Table 3.
Nutrient composition among treatments ration were
significantly different (P<0.05). Substitution of napier
grass by ramie leaves hay on level 20% decreases the
DM weight of ration, but it was still higher than the
DM weight of the control ration. Substitute of napier
grass by silage 20% caused the DM weight of ration
was lower than of the control ration. On higher level
substitute (30% and 40%), DM weight of ration that
was produced were higher than control. The difference
of ration’s DM weight was not only because of hay
and silage alone, but also because of other ingredients
(Table 1).
Ash shows the mineral contents of the substances.
Generally, substitution of king grass with ramie
increased the ash-content. This was because of the
higher ash-content of both preserved ramie leaves
compared to napier grass. The higher ramie hay
and silage on ration, the higher ash-content was.
Substitution of napier grass with ramie leaves hay
increased the ration’s ash-content higher compared to
substitution with silage. This was because of the ash-
content on ramie leaves hay was higher than on ramie
leaves silage (Table 1). Ash-content of ramie leaves
was dominated by Ca that ranging from 4 – 5%. High
content of 6% Ca on ramie leaves was also reported by
January 2011 27
Vol. 2 No. 1 THE EVALUATION OF NUTRIENT QUALITY

Duarte et al. (1997). The high content of Ca on ramie
leaves was expected to be more available for dairy
animals than inorganic Ca that usually added in ration
(McDowell, 2003).
Ration fat-content (EE) that contained both
preserved ramie leaves (silage and hay) were not
different with control. Ration containing 40% silage had
higher EE content than control and ration containing
hay on every level. Because of that, the higher silage
level that was added, the higher EE content was on
ration. On the contrary, the higher hay level added,
the lower EE content was. Crude fat-content on ration
was high because of the high percentage of pressed
coconut cake (Table 2).
Ration containing hay ramie leaves had higher CP
content than control. Hay ramie leaves contained of
14.01% CP were able to increase the CP content of the
ration significantly. Eventhough, it was not obviously
different, ration containing silage ramie leaves had a
higher CP content than control. There was no obvious
different caused by the level of hay-added on CP
content of ration. The low CP content on using silage
was proceed from dried cassava on hay ramie leaves
that has 2.6% CP content. The use of dried cassava on
ensilage had caused ramie ensilage to have content of
10.2% CP, which was not really different with napier
grass (8.9%).
Ration consisted of preserved ramie leaves had a
lower CF content than control. This was because of
lower CF content on preserved ramie leaves compared

Table 1. Ingredients and their chemical composition

DM Ash CP EE CF TDN Ca P
No. Feed Ingredients
------------------------------------------------- (%) -------------------------------------------------------
1. Ramie hay 90.43 21.57 14.02 3.70 13.09 52.79 4.65 2.18
2. Ramie silage 90.10 17.90 10.20 4.41 11.10 62.30 3.98 0.17
3. Napier grass 22.20 12.00 8.69 2.71 32.30 52.40 0.48 0.35
4. Rice bran 87.70 13.60 13.00 8.64 13.90 67.90 0.09 1.39
5. Pollard 88.50 5.90 18.50 3.86 9.80 69.20 0.23 1.10
6. Pressed coconut cake 88.60 8.20 21.30 10.90 14.20 78.70 0.17 0.62
7. Corn 86.80 2.20 10.80 4.28 3.50 80.80 0.23 0.41
8. Dried cassava 79.50 4.70 2.60 7.00 5.70 78.50 0.17 0.09

Table 2. Ingredients and rations composition

P0 P1 P2 P3 P4 P5 P6
Feed Ingredient
------------------------------------------- (%) -------------------------------------------------
Ramie hay 0 0 0 0 20 30 40
Ramie silage 0 20 30 40 0 0 0
Napier grass 50 30 20 10 30 20 10
Rice bran 10 10 12 7.87 10 10 10
Pollard 10.39 17.67 17.85 23.16 15.64 19.74 18.42
Pressed coconut cake 7.32 11.8 13.67 15.09 5 5 5
Corn 18 9.03 5 3 13.09 7.04 5
Dried cassava 3.94 0 0 0 10 10 15
CaCO3 0.35 1 1 0.38 1 0 0
DCP 0 0.5 0.5 0.5 0.28 0 0
TDN 66.5 66.5 66.91 68 66.5 66.5 66.5
PK 12 12 12 12 12 12 12
Ca 0.41 1.518 1.879 2 1.584 1.558 1.982
P 0.561 0.614 0.635 0.589 0.462 0.455 0.436

28 January 2011
DESPAL ET AL. Indon. J. Nutr. and Feed Sci.

to napier grass. and NH3 as the fermentation product from protein.

The higher use of preserved ramie leaves (hay
or silage) in the ration, the lower crude fiber-content
on ration was. The lower crude fiber-content on
ration was expected to cause a higher digestibility.
According to Despal (2000), crude fiber had a negative
correlation to digestibility. The lower crude fiber was,
the higher digestibility of the ration was. But, the very
low crude fiber on dairy animal ration can intrude the
syntheses of milk fat that impacted on the lowering
of milk production. This was because of the low
content of crude fiber deliver the VFA pattern that has
more proportion of molar propionate acid. Propionate
was much more used as energy reserve and a bit as
syntheses of milk fat. Seymour et al. (2005) reported
that the content of milk fat had a negative correlation
with propionate and butyrate content of the diet but
had a positive correlation with acetate.
Fermentability and digestibility
Ration fermentability can be measured by VFA
production as the product of organic matter fermentation
VFA was the main energy source to ruminant livestock
and was an output from the ration fermentation on
rumen (Orskov and Ryle, 1990). On that account, VFA
production in rumen could be used as an indicator
on ration fermentability (Hartati, 1998). VFA profile
(molar proportion of VFA) that yielded could be used
to describe whether a ration was approprioate to the
livestock. The influence of adding ramie leaves silage
and hay on ration fermentability was shown in Table
4. Statistical analysis resulted that organic matter and
protein fermentability of the ration were not showing
any different among treatments (P>0.05).
According to Sutardi (1980), the optimal range of
ration VFA was 80-160 mM. Total VFA that yielded
in this study was so low compared to range of VFA
that was needed for the optimal growth of rumen
microorganism. This was because of the different
measurement method, in case on this research VFA
was measured by GC, whereas on Sutardi (1980), the
measurement was done using steam destilation. The
low values of VFA on measurement using GC were
also found by Despal (2005); Madrid et al., (1999);

Table 3. Nutrient composition of complete ration

Ash EE CP CF
DM
Treatments
(%)
------------------------------------- (% DM) ---------------------------------------
P0 82,17 b
10,01a 4,52abc 12,97a 23,78e
P1 81,23a 10,81a 2,83a 13,69ab 16,97cd
P2 86,15e 11,11b 5,21bc 12,69a 14,29bc
P3 86,14e 11,17b 5,58d 13,16abc 11,45a
P4 85,26e 12,03b 4,75bc 13,83bcd 16,53d
P5 83,92d 14,26c 3,77ab 14,52d 17,18d
P6 83,86c 14,94c 3,94abc 14,31cd 13,08ab
Note: Means in the same column with different superscript differ significantly (P <0.05)

Table 4. Fermentability of complete ration

VFA (mM/L)* NH3
Treatments
Acetate Propionate Isobutyrate Butyrate Isovalerate Total (mM/L)
P0 26,25 4,44 0,62 2,67 0,32 34,3 11,46
P1 26,31 5,47 0,37 3,16 0,31 35,62 10,3
P2 27,74 6,56 0,58 3,75 0,4 39,03 10,62
P3 24,78 6,38 0,39 3,65 0,27 35,47 9,67
P4 22,57 4,57 0,52 2,93 0,15 30,74 8,42
P5 25,27 4,13 0,47 2,37 0,19 32,43 9,7
P6 18,42 3,2 0,31 2,31 0,11 24,35 8,94

January 2011 29
Vol. 2 No. 1 THE EVALUATION OF NUTRIENT QUALITY

and McCullough and Sisk (1972). On steam distillation The increasing of OM digestibility was in line
methods, all volatile substances are counted as VFA, with increasing of DM digestibility. As reported by
but not in VFA measured using GC. Sutardi (1980), because of most components of DM
Ration containing hay was less fermentable were consisted of OM so that factors that influenced
than ration containing silages. This was because DM digestibility, could also influence OM digestibility.
of microorganism activity on the ensilage helped
digesting the feedstuffs and caused silage in the rumen
Table 5. In vitro digestibility of complete ration
system more fermentable. The same result was also
found by Schingoethe et al. (1976). DMD OMD
Acetic acid was present in greatest amount and Treatments
(%) (%)
the proportion of propionic acid usually exceeded
P0 61,21a 60,40a
that of butyric (Balch and Rowland, 1956). Acetate
proportions to total VFA of the respective rations were P1 66,33abc 66,22abc
76.5%; 73.9%; 71.1%; 69.9%; 73.4%, 77.9% and P2 69,53bc 69,25c
75.6%. The use of silage (P1 – P3) gave a lower acetate
proportion than control. The higher use of silage on P3 71,91c 72,33c
ration, the lower acetate proportion . This was because P4 66,81abc 67,44bc
of the lower content of CF in silage containing ration
P5 61,63ab 61,89ab
compare to control (McCullough and Sisk, 1972).
The use of hay on certain level might reduce acetate P6 65,00ab 66,14abc
Note: Means in the same column with different superscript differ
proportion, however not as much as on silage. On the
significantly (P <0.01)
use of hay as much as 30%, acetate proportion was
higher compared to control. The high proportion of
acetate on the use of hay can be found in Esdale et al. Conclusion
(1968).
Ammonia was the main source of nitrogen Ramie leaves silage and hay used as substitute
to synthesize the microorganism’s protein, so its for napier grass may improve nutrient content and
concentration on rumen was a case that had to be ration digestibility. Eventhough ration fermentability
observed (Satter and Slyter, 1974). According to using ramie leaves silage and hay lower than control,
McDonald et al. (2002), the range of NH3 optimal fermentability in all ration still in optimal range.
concentration to synthesize the rumen microorganism’s Acetate proportion was higher on hay ramie leaves
protein was 6 – 21 mM. The NH3 that yielded from substitution though it had a lower digestibility than on
protein fermentation on the experimental rations were silage. Either silage and hay ramie leaves can be used
in optimal range for the growth of livestock and not up to 40% as Napier grass substitute in Etawah TMR.
excessive.
Digestibility was an early indication on the
availability of nutrients in certain feed to livestock ReferenceS
(Yusmadi, 2008). The influence of hay-added and
silage-added to ration on in vitro digestibility is
AOAC. 1999. Official methods of Analysis. AOAC
shown on Table 5. Ration treatment highly influential International. Washington.
(P<0.01) to ration DM and OM digestibility.
Balch, D. A. & S. J. Rowland. 1956. Volatile fatty acids
Ramie leaves silage-added to ration increased the and lactic acid in the rumen of dairy cows receiving a
DM and OM digestibility in line with the increased variety of diets. Nutr. 11 : 288 – 298.
level. The increase in digestibility also observed in Crompack. 1998. Reference Manual CP-9002 Gas
its on hay ramie leaves-added however is was not Chromatograph. Middleburgh. Netherland.
as much as on silage. Moreover 30% hay-added was
Despal. 2000. Kemampuan komposisi kimia dan kecernaan
ration gave a relative similiar digestibility to control. A in vitro dalam mengestimasi kecernaan in vivo. Media
higher digestibility of silage compared to hay was also Peternakan 23 (3): 84 – 88.
found by Yusmadi (2008). Dry matter and organic Despal. 2005. Nutritional Properties of Urea Treated Cocoa
matter pattern was inversely proportional to CF ration. Pod for Ruminant. Cuvillier Verlag. Goettingen.
The higher CF was, the lower digestibility was. This
Despal. 2007. Suplementasi nutrien defisien
case was in mutual according to Despal (2000). untuk meningkatkan penggunaan daun rami
(Boehmeria nivea L. Gaud) dalam ransum

30 January 2011
DESPAL ET AL.
domba. Media Peternakan 30 (3): 181-188.
Despal & I. G. Permana. 2008. Penggunaan berbagai Teknik
Preservasi untuk Optimalisasi Pemanfaatan Daun
Rami sebagai Hijauan Sumber Protein dalam Ransum
Kambing Peranakan Etawah. Laporan Penelitian
Hibah Bersaing. LPPM Institut Pertanian Bogor,
Bogor.
Duarte, A. A, V.C. Sgarbieri & E. R. B. Juniar. 1997.
Composition and nutritive value of ramie leaf flour
for monogastric animals. Revista PAB : 32 (12).
Esdale, W. J., G. A. Broderick & L. D. Satter. 1968.
Measurement of ruminal volatile fatty acid production
from alfalfa hay or corn silage rations using a
continuous infusion isotope dilution technique. J.
Dairy Science 51 (11) : 1823 – 1830.
FAO. 2005. Animal Feed Resources Information System.
htttp://www.fao.org/ag/aga/agap/frg/afris/Data/361.
HTM. [1 Juni 2009].
General Laboratory Procedure. 1966. Departement of Dairy
Science University of Wisconsin.
Hartati, E. 1998. Suplementasi minyak lemuru dan seng ke
dalam ransum yang mengandung silase pod kakao dan
urea untuk memacu pertumbuhan sapi Holstein jantan.
Disertasi. Program Pasca Sarjana. IPB. Bogor.
Madrid, J., M. D. Megias & F. Hernandez. 1999.
Determination of short volatile fatty acids in silages
from artichoke and orange by-products by capillary
gas chromatography. J. Sci. Food Agric 79: 580-584.
McCullough, M. E. & K. R. Sisk. 1972. Crude fiber, form
of ration, type of silage and digestibility of optimum
rations. J. Dairy Sci. 55 (4) : 484.
Indon. J. Nutr. and Feed Sci.
McDonald, P. R. A., Edwards J. F. D. Greenhalge & C. A.
Morgan. 2002. Animal nutrition. 6th Edition. Longman
Science and Technology. New York.
McDowell, Lee Russell. 2003. Minerals in Animal and
Human Nutrition. 2nd Edition. Elseivier Science.
Amsterdam.
NRC. 1981. Nutrient Requirement of Goat. National
Academy Press. USA.
Orskov, E.R and Ryle M. 1990. Energy
Nutrition in Ruminant. Elseivier. London.
Schingoethe, D. J., H. H. Voelker, G. L. Beardsley, & J. G.
Parsons. 1976. Rumen volatile fatty acids and milk
composition from cows fed hay, haylage, or urea
treated corn silage. J. Dairy Sci. 59 (5) : 894 – 901.
Satter, L. and L. L Slyter. 1974. Effect of` amonia
concentration on rumen microbial protein
production in vitro. Br. J. Nutr. 32 : 199 – 208.
Sutardi, T. 1980. Landasan Ilmu Nutrisi. Jilid I. Fakultas
Peternakan. Institut Pertanian Bogor. Bogor.
Seymour, W. M., D. R. Campbella, & Z. B. Johnson. 2005.
Relationships between rumen volatile fatty acid
concentrations and milk production in dairy cows : a
literature study. Animal Feed Science and Technology
119 : 155–169.
Yusmadi. 2008. Kajian Mutu dan Palatabilitas Silase dan Hay
Ransum Komplit Berbasis Sampah Organik Primer
pada Kambing PE. Tesis. Program Pascasarjana.
Institut Pertanian Bogor. Bogor.
January 2011 31
Indon. J. Nutr. and Feed Sci., January 2011, pages 32-35 Vol. 2 No. 1
ISSN 1410-2390

Addition of Chlortetracycline to improve the Performance of broilers fed

Local diets

R. Murwani a*, I. A. Setyawan b, and A. A. K. Ariestab

a
Lab. of Nutritional Biochemistry, Department of Nutrition and Feed Science
Faculty of Animal Science, Diponegoro University Semarang, Indonesia
b
Supervised undergraduate students
(Received 09-11-2009. ; Accepted 14-07-2010)

Abstract

A research was carried out to study the effect of chlortetracycline addition in drinking
water on the performance of broilers fed corn-mungbean base diet. The diet was composed
of corn and mungbean as the major ingredients. One hundred of day-old chicks CP 707
were randomly assigned into two treatments i.e. 1) no addition of chlortetracycline in the
drinking water (D1), (2) addition of chlortetracycline in the drinking water (D1+). Diet
and water were offered ad libitum. Feed consumption, body weight, and feed conversion
were recorded up to day 21. Chlortetracycline addition via drinking water affected
significantly feed consumption and feed conversion (P<0.05). Broilers with chlortetracycline
administration showed lower feed consumption and feed conversion. The results indicated
that chlortetracycline additive can improve the performance of broilers fed corn-mungbean
base diet.

Key words : Chlortetracycline, mungbeans, corn, sorghum, feed conversion

INTRODUCTION broiler production (Murwani and Bayuardhi, 2007).

Subtherapeutic level in-feed antibiotics are
commonly used in broiler commercial feeds (Murwani
and Bayuardhi, 2007). In feed-antibiotic exerts
its effect by decreasing the number of pathogenic
bacteria and hence promoting a better microbial
balance in the gut and improving nutrient utilization
(Dibner and Richards, 2005). They are therefore
useful in preventing bacterial infection and protect
broilers from heat stress in hot and humid climate like
Indonesia. It is also a common practice to administer
antibiotics via drinking water in the management of
*
Coresspondence author
Department of Nutrition and Feed Science, Faculty of Animal Science,
Diponegoro University, Campus drh. Soejono Koewoemowardojo
Tembalang Campus Semarang 50275, Central Java, Indonesia
e-mail : rmurwani@telkom.net
Such practice is helpful in maintaining broilers health,
suppressing mortality, supporting maximal growth and
sustaining profitable bussiness.
In-feed antibiotic is commonly used in commercial
broiler diet based on corn, soybean meal, and meat
bone meal. As these feed ingredients rely heavily on
imports, high price of feed can not be avoided. In
attempt to substitute imported feed ingredients, the
use of locally available grains were studied. Soybean
meal as source of vegetable protein can be replaced
by mungbean which has high protein content among
various local beans. Sorghum can also be used together
with corn as source of energy. Although sorghum has
been known to contain an anti nutritive-tannin, its use
in appropriate amount in the diet composition can be
beneficial to broilers (Nyachoti et al., 1996; Murwani,
2008; Murwani, 2009). Meat bone meal which has
been banned for use in ruminants feed remains an
important feed ingredients for broilers as it supplies
high animal protein. However it can be substituted
by various local animal protein such as fish meal,

32 January 2011
MURWANI ET AL. Indon. J. Nutr. and Feed Sci.

rejected milk, or by products of bakery industries. The Table 1. Composition and nutrient contents of experimental
following research therefore was carried out to study diets.
the effect of antibiotic additive i.e. chlortetracycline Ingredients (%) D1 D1+
in drinking water in broilers fed local feed ingredients
based diet. Treatment of antibiotic in drinking
water - 500 ppm
Corn 36.71 36.71
MATERIALS AND METHODS Mungbeans 29.95 29.95
Sorghum 2.90 2.90
Birds and diets Protein Mix 21.26 21.26
Coconut Oil 2,9 2,9
All feed ingredients were obtained from local
Mineral 3.38 3.38
feed producers. Corn, sorghum and mungbean were
obtained in grain form with moisture content around Baker yeast 2.90 2.90
11%. These feed ingredients were ground separately Total 100 100
and stored in clean water- tight plastic drum until Nutrient Contents
mixed. They were also checked for the presence of Metabolisable Energy (Kkal/kg)2 3230.28 3230.28
mycotoxin, but no mycotoxin was detected. Crude protein3 22.7 22.7
Corn, sorghum, mungbean, protein mix, inactivated Crude fat3 1.09 1.09
bakery yeast (fermipan), vitamin mixture and mineral
Crude fiber3 2.32 2.32
mixture were used to compose the basal diet (Table 1). 1
The contens of mineral mix per kg i.e. 32.5% Ca, 10% P, 6 g Fe,
Protein mix consisted of locally produced milk powder 4 g Mn, 0.075 g , 0.3 g Cu, 3.75 g Zn, 0.5 g vitamin B12, 50000
and albumin powder. Albumin powder was prepared IU vitamin D3
by steaming albumin obtained from side product of
2
Based on calculated values
3
Based on proximate analysis.
local bakery producers, dried and then ground.
A total of 100 Ross CP 707 day old unsexed
broilers with initial body of weight 37± 5 g were chick
used in this experiment. They were offered free access broiler chicken producers (Murwani and Bayuardhi,
to sugar and vitamin mix solution on the arrival. The 2007).
experimental chicks were randomly assigned into 2
groups (in a warm brooder) and given the following Feed consumption, body weight
treatments: 1) without chlortetracycline (D1), (2) with and feed conversion
chlortetracycline via drinking water (D1+) (Table 1).
On day 7, the birds from each large groups were further Feed consumption was determined from the
allocated randomly into 5 replicates with 10 chickens amount of feed given ad libitum minus feed that
in each replicate. was not consumed. Body weight was measured each
The dose of antibiotic used was 500 ppm week with an electronic scale. Feed conversion was
(500 mg/L water). The diet was formulated to meet determined from the ratio of feed consumption to body
nutrient requirement of broilers with protein level on weight.
22.7%. The calculated ME of 3200 Kcal/kg. ME was A completely randomized design with 2 treatments
calculated from ME value of each feed ingredient and 5 replicates was employed. All data were analyzed
(NRC, 2004 and Hartadi et al., 1986). Chicks were using ANOVA, and Duncan’s multiple range test .
offered ad libitum access to the diet and drinking water.
Antibiotic-free vitamin was given through
drinking water and mineral mixture was mixed with
the diet. Birds were vaccinated with commercial ND RESULTS AND DISCUSSION
La Sota (PT. Medion Indonesia) on day-4 via eye
drop and subcutaneous simultaneously. The dose and Feed consumption of broilers in D1 was
vehicle of vaccine was used according to instruction significantly higher than broilers in D1+ (P<0.05)
sheet. Subcutaneous route was given with automatic (Figure 1). Antibiotic supplementation in drinking
injector. The treatments were performed for 25 days water reduced feed consumption.
of starter period, in an open broiler-house with similar
condition as that found in most small to medium scale

January 2011 33
Vol. 2 No. 1 CHLORTETRACYCLINE ADDITIVE

1000 874,06 2
751,40
800
1,5
600

gram/bird
gram/bird

1
400 A B A
A B
200 0,5

0
0
D1 D1+
D1 D1+
Figure 3. Means of feed conversion of broilers as affected
Figure 1. Means of feed consumption of broilers as affected
by antibiotic addition in drinking water.
by antibiotic addition in drinking water (g/bird)

There is no significant effect of treatments on compsution might indicate that antibiotic improved
body weight of broilers at 21 d (p>0.05). Antibiotic the effectiveness of feed utilization which could fulfill
adminstration did not affect body weight of broilers the energy and nutrient needs of broilers. Antibiotic
(Figure 2). might induce satiety system in birds and therefore
had less consumption. Antibiotics might also excert
700 its effect by improving gut morphology, especially on
560,80 small intestine, suppressing pathogenic microbes, and
600 528,80
consequently improving nutrient absorption (Parks et
500 al., 2000; Perez et al., 2005; Murwani, 2008). Such
400 effect was manifested by accretion into lean which is
gram/bird

300 reflected in body weight of broilers. Consequently that

A
B in feed conversion of broilers fed in D1+ was lower
200
than D1. The results of the present study showed
100 that administration of chlortetracycline via drinking
0 water to broiler starter fed corn-mungbean base diet
D1 D1+ improved feed conversion.
Figure 2. Means of broilers body weight of as affected by
CONCLUSION
antibiotic addition in drinking water
Administration of chlortetracycline via drinking
There is a significant effect of treatments on water is beneficial to broilers starter fed corn-mungbean
feed conversion of broilers at 1 – 21 day (p<0.05). base diet.
Antibiotic administration improved feed conversion
(Figure 3). ACKNOWLEDGEMENT
Feed consumption of broilers is affected by
palatability. It is generally accepted that predominat This work was supported by Research Grant
senses of birds including broilers are sight and touch. “Hibah Kompetensi” fiscal year 2008 and 2009 from
Birds have taste buds and receptors, as well as touch Directorate General of Higher Education which was
receptors in the beak. Therefore, birds can taste and granted to the first author.
perceived physical form of the diets including colour,
although with less acuity than mammals (Klasing, REFERENCES
2000). The use of corn and mungbean as major
Dibner, J. J & J. D Richards . 2005. Antibiotic growth pro-
ingredients in the diet give certain colour and taste
moters in Agriculture: History and Mode of Action.
which are due to yellow colour of corn and the green
Poult. Sci., 84:634-643.
skin colour of mungbean. Therefore, the palability of Hartadi, H., S. Reksohadiprodjo & Tillman, A.D. 1986.
diet in birds with and without antibiotic should be the Feed Composition Table for Indonesia. Gadjah Mada
same. However, administration of antibiotic resulted in University Press. Jogjakarta.
a decrease of feed consumption (D1+). Decreasing feed

34 January 2011
Vol. 2 No. 1
Klasing, K.C. 2000. Comparative Avian Nutrition. CABI
Publishing. Cambridge.
Murwani, R. & B. Bayuardhi. 2007. Broilers serum
cholesterol and glutamic oxaloacetic transaminase
and their relation to antibiotic in feed and medication
programs in four broiler producers in Semarang
region-central Java, Indonesia. Intl. J. of Poult. Sci.,
6(4): 266-270.
Murwani, R. 2008. Aditif Pakan. Aditif Alami Pengganti
Antibiotika. Unnes Press, Semarang.
Murwani, R. 2009. Effect of Mung bean as Local Feed
Ingredients to Substitute Soybean Meal in the Diet
on the Performance of Broilers. The first International
Seminar on Animal Industry: Sustainable Animal
Production for Food Security and Safety. Bogor.
Nyachoti, C. M., J. L. Atkinson & S. Leeson. 1996. Response
of broiler chicks fed a high-tannin sorghum diet. J. of
App. Poult. Res. 5: 239¬-245.
THE EVALUATION OF NUTRIENT QUALITY
NRC. 1994. Nutrient Requirements of Poultry. National
Academy Press. Washington, D.C.
Parks, C.W., J. L. Grimes, P. R. Ferket & A. S. Fairchild.
2000. The case for mannanoligosaccharides in poultry
diets. An alternative to growth promotant antibiotics?
Proc. of Alltech’s 16th Annu. Symp. (Lyons, T.P.
and Jacques, K.A. eds.). Nottingham Univ. Press,
Nottingham UK., p.45-60.
Perez, A.R., E. Amit-Romach & Z. Sklan Dand Uni. 2005.
Mucin dynamics and microbial populations in chicken
small intestine are changed by dietary probiotic and
antibiotic growth promoter supplementation. J. Nutr.
135:187-192.
January 2011 35
Indon. J. Nutr. and Feed Sci., January 2011, pages 36-41 Vol. 2 No. 1
ISSN 1410-2390

Efficacy of Methionine Addition in Laying Hens Fed Corn-Soy-Palm Kernel

Meal Based Diet

M. Ridla*, Sumiati, J. Jachja, T. Toharmat, I. G. Permana and Nahrowi

Department of Nutrition and Feed Technology, Faculty of Animal Science, Bogor Agricultural University
Jln. Agatis, Kampus IPB Darmaga Bogor 16680
(Received 09-11-2009 ; Accepted 02-08-2010)

ABSTRACT

The objective of this study was to investigate the effect of methionine supplementation either
in drinking water or diet on feed conversion ratio, hen day egg production from hens age
22-24 weeks, egg production, egg weight, percentage of egg yolk, egg albumin, eggshell
weight, as well as eggshell thickness in laying hens fed corn-soy-palm kernel meal based
diet. Three hundreds and fifty commercial laying hens age 16 weeks with initial weight of
1626.7 ± 38.7 g were assigned randomly in a completely randomized design to seven dietary
treatments. The treatments were as follows: (S0) deficient in methionine without addition of
methionine; (S1, S2, S3) S0 diet with addition of methionine in the diets at level of 0.05, 0.10,
and 0.15 giving total methionine content in the rations of 0.33, 0.38, and 0.43%; (S4, S5, S6)
S0 diet with addition of methionine in the drinking water at level of 0.025, 0.05, and 0.075%.
Diets and water were offered ad libitum. Data were analyzed statistically using analysis of
variance (ANOVA) according to the procedure of SAS. The average intake of methionine of
S2 during four weeks of the experiment was 388.8 mg/day/bird, while the average intake of
methionine of S0 and S1 were less than the requirement, i.e., 276 mg/day/bird and 330 mg/
day/bird, respectively. The intake of methionine of S3, S5, and S6 was over the requirement
(432 mg/day/bird) and this amount of methionine could decrease the performances of the
hens. Supplementation of 0.1% methionine in the diets (S2, total methionine in the diet
was 0.38%), and 0.05% in the drinking water gave the best results in terms of hen-day egg
production, feed conversion ratio, egg weight, albumin weight, and shell weight. It is
concluded that methionine should be added to corn-soy-palm kernel based poultry diets as
much as 0.1% in diet and 0.05% in the drinking water.

Key words: Palm kernel meal, laying hens, methionine, egg, albumin, shell

INTRODUCTION Adoption of low protein diet and supplementation

Corn-soy-palm kernel meal based poultry diets are
deficient in limiting amino acids such as methionine.
Amino acid balance and nitrogen retention in poultry
diets are improved by methionine supplementation.
*
Coresspondence author
Department of Nutrition and Feed Technology
Faculty of Animal Science, Bogor Agricultural University
Agatis street, Kampus IPB Dramaga Bogor 16680
e-mail : hm_ridla@yahoo.com
of synthetic amino acid in poultry diets recently is
becoming relevant in feed formulation to minimize
the nitrogen excretion and production cost. Levels
and balance of amino acids in the diets are important
nutritional variables that affected feed efficiency in
layer (Al-Saffar and Rose, 2002).
Liquid methionine hydroxy analogue and solid
DL-Methionine (DLM) product are available in the
market. Efficacy of the products is believed to be
correlated with nutritional value of the diet, climate
and the type of poultry. Supplementation of 0.25 %

36 January 2011
RIDLA ET AL. Indon. J. Nutr. and Feed Sci.

DL-methionine in starter diets and 0.20% in finisher egg quality in terms of the percentage of egg yolk, egg
diets in the corn-soya based diet increased performance albumen, eggshell, as well as eggshell thickness.
of broiler (Jahja et al,. 2007). However, the optimum Data were analyzed statistically using analysis of
level of methionine addition in broiler may differ from variance (ANOVA) (Steel and Torrie, 1991).
that in layer, and moreover, inclussion of palm kernel
meal (PKM) in laying hens diets may affect nutrient
retention including methionine. Table 1. Formula and chemical composition of experimental
The objective of this study was to investigate basal diets
the effect of methionine supplementation either in
Ingredients (%) Chemical Composition
drinking water or diet on egg production and its quality
Nutrient (%)
in laying hens fed corn-soy-PKM based diet.
Local Corn 53.96 Dry matter 86.16
Indian SBM 18.08 Ash 10.18
MATERIALS AND METHODS Limestone Pow 9.0 Crude protein 15.25
Rice Bran 7 Crude Fiber 5.45
Three hundred and fifty ISA-Brown strain CGM USA 2.74 Ether extract 2.66
laying hens, 16 weeks of age, were purchased from Nitrogen free
commercial breeder. Liquid methionine hydroxy Crude Palm Oil 2.13 extract 52.62
analogue and solid DL-Methionine (DLM) was Dicalcium
supplied by Sumitomo Chemical Co., Ltd. Feedstuffs phosphat 1.28 Ca 4.38
were obtained and mixed in a small commercial feed Sodium
mill in Bogor-Indonesia. The pullets were kept in Bicarbonat 0.29 P 0.99
individual cages in conventional housing system (an Salt 0.20 NaCl 0.1
open-side poultry housing). Each cage was completed L-Lysine 0.11 GE 3984
with feeder and drinking water. The lighting was Choline Chloride 0.10 EM (kcal/kg) 2868*
provided for 16 hours from 05:00 AM to 21:00 PM
Mineral mix 0.05
daily.
Vitamin mix 0.05
Experimental rations were formulated based on
*Calculated from 0.72 x Gross Energy
poultry requirement according to NRC (1994) using
corn-soy-PKM based diet. The formula and chemical
composition are shown in Table 1. The ration was
mixed twice a month. RESULTS AND DISSCUSSION

Experimental design Response of feed intake

Three hundreds and fifty commercial laying hens
age 16 weeks with initial weight of 1626.7±38.7 g
were assigned randomly in a completely randomized
design to seven dietary treatments. The treatments were
as follows: (S0) basal diets deficient in methionine
without addition methionine; (S1, S2, S3) S0 basal
diets with addition of methionine in the diets at level
of 0.05, 0.10, and 0.15 giving total methionine content
in the rations of 0.33, 0.38, and 0.43%; (S4, S5, S6) S0
basal diets with addition of methionine in the drinking
water at level of 0.025, 0.05, 0.075%. Diets and water
were offered ad libitum.
Feed and water intake, egg production, and egg
weight were recorded daily. Feed convertion ratio,
henday egg production on the period 22-24 week
of egg, and methionine intake were then calculated.
During the last days of the experimental period, eggs
sample from each treatment were taken to determine
The means feed intake of the laying hens are
presented in Table 2. Supplementation of DL-
methionine either in the diets or in the drinking water
did not affect the feed intake of the hens. However,
the average of feed intake of the hens offered diet
supplemented with methionine either in diet or drinking
water were higher than that of the hens offered control
diet (S0), except for the hens fed S6 diet ( + 0.15%
DL-methionine in the drinking water). This feed
intake was similar to that reported by Bell and Weaver
(2002) that feed intake of brown egg layers at 22 weeks,
23 weeks, and 24 weeks old were 99.1 g/hen/day,
101.4 g/hen/day, and 103.2 g/hen/day, respectively.
Response of water intake
The means of water intake of the laying hens
are presented in Table 3. Supplementation of DL-

January 2011 37
Vol. 2 No. 1 STUDY ON EFFICACY

methionine either in the diets or drinking water Table 3. As a response to DL-methionine supplementation
decreased the average of water intake during the first means of on water consumption (ml/day/bird)*
five weeks of laying period. Compared to the water
Age (Weeks)
intake of the hen fed control diet, the decreasing water Treatments
intake were 16.04% (S1), 13.16% (S2), 20.58% (S3), 22 23 24
14.82% (S4), 17.78%, and 27.71% (S6). The hens S0 298.54±12.99 316.74±13.47 361.11±35.92
fed S6 diet (+ 0.15% DL-methionine in the drinking S1 244.54±26.28 263.63±8.59 298.67±51.07
water) had the lowest water intake. The reduction in S2 264.97±17.80 265.14±19.78 279.76±46.95
water intake could be due to the taste of water as the S3 256.03±16.42 235.43±13.61 250.90±34.94
result of rather high content DL-methionine. Water S4 249.86±14.14 266.06±15.36 293.41±48.01
intake recommended by Bell and Weaver (2002) in
S5 248.80 ± 2.80 258.91±22.06 278.46±25.17
laying hens was 220 ml/hen/day at house temperature
of 28.9oC and 300 ml/hen/day at 34.4oC. S6 216.89±21.33 228.17±17.07 49.51±29.20
* S0= basal diet without DL methionine supplementation; S1=
basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
Table 2. As a response to DL-methionine supplementation methionine in the diet; S4= basal diet +0.025% DL-methionine
on means of feed consumption (g/day/bird)* in the drinking water; S5= basal diet + 0.05% DL-methionine in
the drinking water; S6= basal diet + 0.075% DL-methionine in
Treatments Age (Weeks) the drinking water.
22 23 24
S0 96.06 ± 2.71 97.33±2.73 103.27± 5.65
0.1% DL-methionine in the diet) yielded the highest
S1 96.14 ± 3.63 97.40 ±3.40 108.36±1.87
egg weight. The eggs weight were almost the same to
S2 98.75 ±1.29 99.80±1.03 112.33±2.63 the standard egg weight suggested by Bell and Weaver
(2002) who reported that the weight of commercial
S3 95.96 ± 3.82 97.12±2.41 109.79± 1.32
egg was 47.5 g/egg (at 22 weeks of age), 49.0 g/egg
S4 94.53 ± 3.69 97.41±2.79 106.63±4.01 (at 23 weeks of age), 50.7 g/egg (at 24 weeks of age),
S5 97.77 ± 2.35 95.85±4.42 106.68± 0.36 and 52.3 g/egg (at 25 weeks of age).
S6 94.90 ± 2.52 94.72±7.90 105.12±1.57
* S0= basal diet without DL methionine supplementation; S1=
Response of egg production
basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
Means of henday egg production are presented in
methionine in the diet; S4= basal diet +0.025% DL-methionine
in the drinking water; S5= basal diet + 0.05% DL-methionine in Table 5. There was no different in henday production
the drinking water; S6= basal diet + 0.075% DL-methionine in in the first five day production period. Henday egg
the drinking water. production at the first week of the production period

Response of egg weight Table 4. Eggs weight (g) of layer with and without
methionine supplementation*
Means of egg weight are presented in Table 4. Treatments Egg Weight
Supplementation of DL-methionine either in the S0 51.24 ± 0.42
diets or drinking water increased the egg weight by of
S1 50.96 ± 0.53
3.22% (S1), 5.2% (S2), 3.45% (S3), 4.96% (S4), 5.10%
S2 51.98 ± 1.03
(S5), and 3.22% (S6). Leeson and Summers (2005)
reported that apart from manipulating feed intake, egg S3 51.62 ± 0.76
size could be manipulated by adjusting dietary levels S4 52.14 ± 0.84
of energy, fat and linoleic acid, or by adjustment the S5 51.69 ± 1.51
levels of protein, methionine and basal sulphur amino S6 51.56 ± 0.89
acids (TSAA). There was a consistent linear trend in * S0= basal diet without DL methionine supplementation; S1=
the increased in egg weight of young birds as TSAA basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
increased from 0.65 to 0.81%. Egg size of young
methionine in the diet; S4= basal diet +0.025% DL-methionine
layers increased by 0.7g for each 0.05% increased in in the drinking water; S5= basal diet + 0.05% DL-methionine in
dietary TSAA. Dietary TSAA was 0.56% (S0), 0.61% the drinking water; S6= basal diet + 0.075% DL-methionine in
(S1), 0.66% (S2), and 0.71% (S3). The hens fed S2(+ the drinking water.

38 January 2011
RIDLA ET AL. Indon. J. Nutr. and Feed Sci.

ranged from 17.36 to 40.88%. During the first five
weeks, all groups of laying hen indicated the similar
trend in hen day production. Although the methionine
supplementation either in solid or liquid form had
no significant effect on henday production, the
supplementation tended to improve the productivity
of layer. The excess of supplemented methionine
tended to reduce henday production.
Supplementation of DL-methionine either in the
diet or drinking water increased henday egg production
by 5.2% (S1), 19.37% (S2), 19.28% (S3), 18.49%
(S4), and 9.94% (S5). However, the supplementation
of methionine in the drinking water at the level of
0.15% (S6) decreased the henday egg production by
8.85%. Increased henday egg production could be due
to methionine role in protein synthesis in the liver of
hens, resulting in increased the follicle formation. The
henday egg production of S3 and S4 at 24 weeks of
henday egg production old of age was 88.48%, and
Table 5. Henday egg production (%) of layer fed diets with
and without methionine addition*
Treatments Age (Weeks)
22 23 24
S0
43 ± 14.52 63.00 ±16.95 68.04 ± 7.45
S1 46 ± 20.06 62.72 ±18.08 75.60 ± 12.52
S2 60 ± 16.55 70.00 ±16.54 81.48 ± 10.77
S3 57 ± 6.96 71.96 ±11.90 83.72 ± 8.82
S4 60 ± 23.42 72.52 ±16.46 80.36 ± 10.50
S5 56 ± 6.49 65.24 ±12.07 70.84 ± 9.37
S6 37 ± 5.65 56.00 ±12.60 69.44 ±15.51
* S0= basal diet without DL methionine supplementation; S1=
basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
methionine in the diet; S4= basal diet +0.025% DL-methionine
in the drinking water; S5= basal diet + 0.05% DL-methionine in
the drinking water; S6= basal diet + 0.075% DL-methionine in
the drinking water.
Response of feed conversion ratio
Feed conversion ratio of laying hen receiving
rations supplemented with methionine are presented
in Table 6. Supplementation of DL-methionine in the
diets and drinking water decreased feed conversion or
increased feed efficiency. The results indicated that
basal diet (S0) had mild deficient of methionine. Pesti
et al., (2005) reported that feed efficiency was reduced
when imbalanced proteins were given to the birds.
Feed conversion decreased with the progress of
the production period. Variation of feed conversion
was high during the first three week of laying period,
especially in weeks 22, 23, 24. The values ranged
from 2.47 to 2.96 in the week 4 of the laying period.
Supplementation of methionine tended to decrease
feed conversion ratio during the first five week of
the production period. The excess of supplemented
methionine tended to increase feed conversion.
Available methionine in the experimental rations did
not meet the requirement of the experimental laying
hen. The supplementation of methionine at level up
to 0.1% corrected the defficiency of methionin in the
ration. However the methionine supplementation up to
0.15% resulted in excess of methionine or imbalance
in dietary amino acid.
Table 6. Means of feed conversion ratio of layers receiving
diets with and without methionine addition*
Age (Weeks)
Treatments
22 23 24
S0 8.87±9.36 5.16±3.02 2.91± 0.50
S1 5.3 ±2.74 4.67±1.74 2.69±0.54
S2 3.44±0.90 4.15±1.28 2.54±0.33
S3 3.38±0.36 3.84±0.65 2.44± 0.24
S4 3.54±1.46 3.79±1.17 2.47±0.24
S5 3.48±0.56 4.15±0.93 2.79± 0.33
S6 8.87±9.36 5.16±3.02 2.91± 0.50
* S0= basal diet without DL methionine supplementation; S1=
basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
methionine in the diet; S4= basal diet +0.025% DL-methionine
in the drinking water; S5= basal diet + 0.05% DL-methionine in
the drinking water; S6= basal diet + 0.075% DL-methionine in
the drinking water.
Response of egg quality
Egg quality of laying hen receiving rations
supplemented with methionine are presented in Table
7. Supplementation of DL-methionine in the diets and
drinking water did not affect the yolk weight, but it
increased the albumin weight. This results showed
that methionine supplementation increased the protein
synthesis in term of albumin protein. Pesti et al.,
(2005) reported that the essential, amino acids were
needed to make protein and other compound. Amino
acid needs change when protein synthesis changes.
When birds were actively producing feather or egg,
for instance, their requirements reflect the amino acids
in these products as well as those needed for tissue
growth and maintenance.
Pesti et al. (2005) reported that the most important
factor in determining egg size was the size of yolk,
largely lipids, and the second most important factor
in determining egg size was the albumen, almost

January 2011 39
Vol. 2 No. 1 STUDY ON EFFICACY

entirely protein. The used of laying hens on protein Table 8. Eggs yolk and eggshell characteristic in laying
and amino acids to synthesis albumen were high, any hens receiving diets with and without methionine
lack of protein resulted in a decrease amount in the of addition*
albumen, and consequently egg size even though the Treatments Parameters
quantity of yolk may be similar. Increasing the protein Yolk Score Eggshell thickness
and amino acid contents of the diet had a marked effect (mm)
on increasing egg size, particularly when the hen laid
small eggs. S0 9.50 0.34
Supplementation of DL-methionine in the diets S1 9.80 0.36
and drinking water increased the egg shell weight, S2 9.50 0.35
except for the S4 and S5. This results indicated that S3 9.70 0.35
there was an increased calcium retention into the egg S4 9.50 0.37
shell due to increased in protein synthesis as methionine S5 9.70 0.37
supplementationed calcium might be bound to protein
S6 10.20 0.36
in the enterocytes and was transported in the blood in *S0= basal diet without DL methionine supplementation; S1=
both ionized and protein-bound forms. basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
methionine in the diet; S4= basal diet +0.025% DL-methionine
Table 7. Eggs components weight in laying hens receiving in the drinking water; S5= basal diet + 0.05% DL-methionine in
with and without methionine addition* the drinking water; S6= basal diet + 0.075% DL-methionine in
the drinking water.
Treatments Parameters
Weight Yolk Albumin Eggshell day/bird, respectively. The intake of methionine by
(g) weight weight weight layers in S3 group was 21.35 % over the requirement
(%) (%) (%) (432 mg/day/bird) and this amount of methionine
S0 51.81 22.56 63.67 10.87 intake could decrease the performances of the hens.
S1 51.84 21.97 64.52 11.29 This result indicated that there was a mild excessive of
S2 51.90 22.00 64.47 11.43 methionine in the S3 diet.
S3 51.96 22.38 64.37 11.01
Supplementation methionine 0.05 % in the
drinking water yielded a better performances compared
S4 52.19 22.44 64.52 10.84
to others (S5 and S6). The methionine intake by layers
S5 51.92 22.67 64.19 10.93 in S5 and S6 group could be exceeded the requirement
S6 52.09 22.53 63.79 11.08 of the hens. The methionine intake of S6 group was
*S0= basal diet without DL methionine supplementation; S1= 161.97% above the requirement, and it could be
basal diet +0.05% DL-methionine in the diet; S2= basal diet +
0.1% DL-methionine in the diet; S3= basal diet + 0.15% DL-
excessive to the birds.
methionine in the diet; S4= basal diet +0.025% DL-methionine
in the drinking water; S5= basal diet + 0.05% DL-methionine in CONCLUSION
the drinking water; S6= basal diet + 0.075% DL-methionine in
the drinking water. Supplementation of 0.1% methionine in the corn-
soy-palm kernel meal based diets which was equal
with 0.38% of the total dietary methionine, or 0.05%
Supplementation of 0.1% methionine in the diets in the drinking water with the same formula of the diet
(S2, total methionine in the diet was 0.38%) resulted resulted in the best performances of young laying hens
in the best performances in terms of hen-day egg in terms of hen-day egg production, feed conversion
production, feed conversion ratio, egg weight, albumin ratio, egg weight, albumin weight, and shell weight.
weight, and shell weight. It could be due to fulfillment It is recomended that methionine should be added to
of the methionine requirement of the hens. The corn-soy-palm kernel based poultry diets as much as
estimated methionine requirement for egg number and either 0.1% in diet or 0.05% in the drinking water.
weight were 364 mg/day/bird and 356 mg/day/bird,
respectively. The average intake of methionine by REFERENCES
layer in S2 group during five weeks of the experiment
was 388.8 mg/day/bird, while the means intake of Al-Saffar, A. A. & S. P. Rose. 2002. The response of of
laying hens to dietary amino acids. World Poult. Sci.
methionine of those in S0 and S1 group were less than
58 : 209-234.
the requirement, i.e., 276 mg/day/bird and 330 mg/

40 January 2011
RIDLA ET AL.
Bell, D.D. & W. D. Weaver, 2002. Commercial Chicken
Meat and Egg Production. 5th ed. Nework, Springer.
Jachja, J., N. Ramli, M. Ridla, Sumiati & T. Toharmat. 2007.
The effectiveness of DL-Methionine suplementation
in diet on performance of starter and finisher broil-
ers. Proceeding of Association of Nutrition and Feed
Scientiest Seminar Gadjah Mada University, July 26-
27th 2007.
Leeson, S., and J.D. Summers,. 2005. Commercial Poultry
Nutrition. 3rd Ed. University Book. Guelph, Ontario,
Canada.
Indon. J. Nutr. and Feed Sci.
National Research Council. 1994. Nutrient Requirements
of Poultry. 9th Revised Edition. National Academy
Press. Washington D. C. USA.
Pesti, G.M., R. I. Bakalli, J. P. Driver , A. Atencio, & E. H.
Foster. 2005. Poultry Nutrition and Feeding.Trafford
Publishing. Canada.
Steel, R. G. D. & J. H. Torrie. 1991. Prinsip dan Prosedur
Statistik. Suatu Pendekatan Biometrik. Terjemahan:
M. Syah. P.T. Gramedia, Jakarta.
January 2011 41
Indon. J. Nutr. and Feed Sci., January 2011, pages 42-45 Vol. 2 No. 1
ISSN 1410-2390

The Effect of Earthworm Supplementation in the Ration on Growth Performance,

Carcass Production, and Abdominal Fat of Broiler

N. D. Dono*, R. Damanik, J. Pasaribu, and A.Wibowo

Faculty of Animal Science, Gadjah Mada University,
3rd Fauna Street – Bulaksumur, Yogyakarta 55281
(Received 09-11-2009. ; Accepted 19-08-2010)

ABSTRACT

Earthworms (Lumbricus rubellus) can be used as an alternative feedstuff in broiler ration

due to its high nutritive values. This study was carried out to investigate the effects of
earthworm meal supplementation in the ration on growth performance, carcass production,
and abdominal fat of broiler. Sixty unsexed DOC strain of Lohmann were randomly
kept within 3 levels of feeding trial, i.e.: LR-0 (basal ration without earthworm meal
supplementation), LR-1.5 (basal ration with 1.5% earthworm meal supplementation), and
LR-3.0 (basal ration with 3.0% earthworm meal supplementation). The basal ration was
composed of : yellow corn, soybean meal, fish meal, wheat pollard, rice bran, coconut oil,
bone meal, and mineral premix. The rations were designed iso-caloric (2979.58 + 18.05 kcal/
kg) and iso-protein (22.36 + 0.40 %). Feed consumption, average daily gain, and FCR of
the broilers were recorded weekly. At 42 days old, chickens were slaughtered to measure
the carcass production and abdominal fat. Data were statistically analyzed with oneway
of ANOVA and followed by Duncan’s new Multiple Range Test for significant differences.
Results showed that supplementation of 3.0% earthworm meal in feed improved FCR
(P<0.05), increased protein consumption (P<0.01), energy consumption (P<0.01), average
daily gain (P<0.05), slaughter weight (P<0.05), and reduced abdominal fat levels (P<0.05),
but did not affect significantly on feed consumption and carcass production.

Key words : Earthworm, broiler chickens, growth performance, carcass production, abdominal fat

INTRODUCTION and will support enhancement of poultry productivity.

Broiler’s meat is a high quality protein source that
can be found easily everywhere. It is a popular source
of animal protein, since broiler meat was inexpensive,
tender, and less fat. These may be the reason why
broiler’s meat popular for human food.
A good quality broiler’s meat is produced by a
quality chicken that reared and given high quality of
feed. Supply of high quality feedstuffs will be useful
*
Coresspondence author
Faculty of Animal Science, Gadjah Mada University,
3rd Fauna Street – Bulaksumur, Yogyakarta 55281
e-mail : nanungdd@yahoo.co.uk
In order to improve productivity, it is important to
supply high quality feedstuffs, which were easy to
access, nutritious, less cost, and can be found locally.
One of the alternative feedstuff that can be explored is
the earthworm. It can be collected from many places
in Indonesia.
Earthworm is a member of annelids that can be
used as an alternative of poultry feedstuff due to its
high quality of nutrient, easy to get, easy to grow,
easy to reproduce, inexpensive, and easy to manage
(Mudjiman, 2004). Rahman (2008) observed that
earthworm meal contains crude protein of 60-72%,
energy of 900-4100 kcal/kg, crude fat of 7-10%, and
ash of 8-10%. Chumaidi (2005) also reported that
earthworm contains crude protein of 52.17%, moisture
of 79.39%, crude fat of 13.86%, and ash of 17.32%.

42 January 2011
DONO ET AL.
Rukmana (1999) reported that earthworm is suitable
as an alternative feedstuff for poultry since it contains
high protein and amino acids. It is also contains
protein and amino acids that are easily digested and
absorbed by the gut. On the other hand, earthworm
also contains anti-bacterial properties (Smith, 2001)
that can reduce pathogenic bacteria of gram negative,
such as : Salmonella typhii, etc. (Waluyo, 2008); non-
toxic (Mambo, 2006) and contains α-tocoferol (vitamin
E), which can be used as anti-oxidant (Purwakusuma,
2007).
In the bio-pharmacy practices, earthworm
is known to have many purposes, such as : anti
oxidative, hepatoprotective (Balamurugan et al.,
2008), antipyretic, antihypertensive, anti-allergic,
anti asthmatic, anticancer, antimicrobial, and anti
inflammatory effects (Ismail et al., 1992; Cooper, 2005;
Prakash et al., 2007; Balamurugan et al., 2007).
Lumbricin, an active compound in earthworm’s,
can be used in poultry feed as immunosuppressant
(Damayanti et al., 2009), resulting in optimizing
daily growth. Resnawati (2008) recommends to use
earthworm meal as supplement in broiler ration.
A further study must be done properly to explore
hidden potencies of earthworm meal in broiler
ration, particularly its effects on growth, and carcass
composition.
Materials and Methods
This research was carried out at the Poultry Unit,
Department of Animal Nutrition and Feed Science,
Faculty of Animal Science, Gadjah Mada University,
Yogyakarta. Materials that used in this study were:
DOC broilers, base diet, and earthworm meal. Sixty
(60) unsexed DOC broilers strain of Lohmann
were used in this study. The birds were plotted into
3 treatment levels, i.e : LR-0 (base diet without
earthworm supplementation; as control diet), LR-
1.5 (control diet supplemented with 1,5% earthworm
meal), and LR-3.0 (control diet supplemented with
3.0% earthworm meal). Every treatment was replicated
4 times, with 5 broilers in each replicate. The control
diet was composed of : yellow corn, soybean meal,
fish meal, wheat pollard, rice polishing, coconut oils,
bone meal, and mineral premix. All diet was design
iso-caloric (2979.58 + 18.05 kcal/kg) and iso-protein
(22.36 + 0.40 %). Earthworm which used was is the
Lubricus rubellus that had been sun-dried for 2-3 days
and grilled properly.
During the feeding trial, total feed consumption,
Indon. J. Nutr. and Feed Sci.
average daily gain, and feed conversion ratio of the
broilers were recoded weekly. At 42 days old, the
broilers were slaughtered and the carcass percentage,
and abdominal fat levels were measured. Variables
that observed in this study were: growth performance
(feed consumption, average daily gain, and feed
conversion ratio), carcass percentage (slaughter weight
and percentage of slaughter weight), and abdominal
fat levels (abdominal fat weight and percentage of
abdominal fat) of broiler.
All data were statistically analysed by the One
Way of ANOVA and followed by the Duncan’s new
Multiple Range Test (DMRT) for significant differences
between treatments.
RESULTS AND DISCUSSION
Growth performance
Growth performance of broilers used in this
study was presented in Table 1. Supplementation of
earthworm meal increased protein intake and energy
intake significantly. Earth worm supplementation
increased protein intake (P<.05) of the birds from 2.24
g/day (LR-0) to 2.41 g/day (LR-3.0). Supplementation
also increased energy intake (P<.05) of the birds from
235.74 kcal/kg/day (LR-0) to 260.43 kcal/kg/day (LR-
3.0), while supplementation at that level did affect
on total feed intake of the birds. Total feed intake of
broilers fed control diet was 83.37 g/day. It was not
different with those that given 1.5% supplementation
(82.89 g/day) or of those given 3.0% supplementation
(84.98 g/broiler/day).
Data on Table 1 showed that on LR-3.0, energy
intake and protein intake increased simultaneously.
This made total feed consumption quite stable and did
not detected significantly. Zuprizal (2006) reported
that total feed consumption of broiler was depended on
energy which is consisted in the ration. Improvement
of that energy and protein consumption was caused
by presence of nutrient that is consisted in earthworm
meal. Therefore, earthworm supplementation increased
protein consumption. It was not different with
Mudjiman’s statement (2004) that since earthworm
meal is so easy to digest, it will easy also to be absorbed
by pores within gastrointestinal walls.
The average of daily weight gain of the birds fed
control diet was 42.46 g/day. Supplementation of 3.0%
earth worm meal increased average of daily weight
gain (P<0.05) from 42.46 g/day (LR-0) to 46.47%
(LR-3.0). On the other hand, feed conversion ratio
(FCR) of the birds fed control diet was 1.96. This FCR
January 2011 43
Vol. 2 No. 1 THE EFFECT OF EARTHWORM

Table 1. Growth performance of broilers during 42 days study

Variabels Level of supplementation
LR-0 LR-1.5 LR-3.0

Total Feed Intake (g/day) 83.37 82.89 84.98

Protein Intake (g/day) 2.24a 2.28a 2.41b
Energy Intake (kcal/kg/day) 235.74a 251.82a 260.43b
Average of Daily Weight Gain (g/day) 42.46a 43.79ab 46.47b
Feed Conversion Ratio (g/g) 1.96b 1.95ab 1.83a
Note : Means in the same row with different superscript differ significantly (P <0.05)

reduced significantly (P<.0.05) up to 1.83 following
by 3.0% earthworm meal supplementation.
This may be caused by the higher amino acids
content of earth worm – especially methionine and
lysine – and fatty acids in the supplemented diet. Both
amino acids and fatty acid contained by the earth worm
more or less will contribute in the growth of the birds
(Rukmana, 1999). Furthermore, earth worm meal
also has strong anti-bacterial properties. Reducing
certain number of enteropathogens alters microbial
colonization in the gut (Rodriguez et al., 1996; Smith,
2001), resulted more nutrient available in the gut.
Therefore, 3.0% earthworm meal supplementation
improved performances of 42 day old broilers.
Carcass percentage
Results showed in Table 2 that slaughter weight
of broilers fed with 3.0% earthworm meal increased
significantly (P<.05) from 1830.75 g/bird (LR-0)
to 1995.55 g/bird (LR-1.5). Addition of 1.5% earth
worm meal on LR-3.0 didn’t automatically increase
slaughter weight (P>.05). Damayanti et al., (2009)
cited Liu et al. (2004) stated that earthworm contains
Lumbricin which have an immuno-stimulation effect.
This active compound is reported to accelerate the
growth rate of the broiler. This might be also caused by
its higher nutrient intake. Soeparno (1994) described
that acceleration of slaughter weight and the whole
growth rate was much depends on the nutrient intake
of the birds.
Data presented in Table 2 showed that carcass
weight was not affected by 3.0% earth worm
supplementation. Carcass weight of the birds fed
control diet was 1139.25 g/bird, while in the LR-1.5
group was 1244.92 g/bird and in the LR-3.0 group was
1311.08 g/bird. At the other hand, carcass percentage
of the bird fed 3.0% earth worm meal was 63.53%,
while in the LR-0 which fed control diet was 62.30
%. Statistical analyses showed that there was no
significant difference among the treatment groups.
Abdominal fat levels
Data presented in Table 2 showed that abdominal
fat levels of the control birds (LR-0) were 20.75 g/bird.
The abdominal fat decreased significantly (P<.05)
to 18.08 g/bird (LR-3.0) following 3.0% earthworm
meal supplementation. However, supplementation of
1.5% earth worm meal did not affect the abdominal
fat weight.
Percentage of abdominal fat of 42 days old birds

Table 2. Carcass production of 42 days old birds and abdominal fat levels fed earthworm meal supplement
Level of Supplementation
Variables
LR-0 LR-1,5 LR-3,0
Slaughter Weight (g/bird) 1830,75a 1995,55a 2084,67b
Carcass Weight (g/bird) 1139,25 1244,92 1311,08
Carcass Percentage (%) 62,3 62,29 63,53
Abdominal Fat Weight (g/bird) 20,75 a
19,58a
18,08b
Percentage of Abdominal Fat (%) 1,13b 1,00b 0,87a
Note : Means in the same row with different superscript differ significantly (P <0.05)

44 January 2011
DONO ET AL.
Earthworm meal is one of high quality protein
source which contains 52.17% to 60-72% crude pro-
tein (Rahman, 2008; Chumaidi, 2005). Previous study
showed that the higher earthworm meal supplementa-
tion level, the more the protein content in the ration.
On the other hand, earthworm meal can be digested
and absorbed easily. As a result, protein metabolism in
the body performed properly, resulting less fat deposi-
tion. It is in the line of Zuprizal (1993) which reported
that increasing protein intake reduced abdominal fat
level. This is also in agreement with the result of Re-
snawati (2004) which showed a high correlation be-
tween feed protein levels with abdominal fat deposi-
tion in broilers. This result was not different with the
result reported by Rezaei et al. (2004).
CONCLUSION
Supplementation of 3.0% earth worm meal in
feed improved feed conversion ratio, increased pro-
tein consumption, energy consumption, average daily
gain, slaughter weight, and reduced abdominal fat lev-
els, but did not affect feed consumption and carcass
production.
Further study on broiler chickens is suggested to
observe the effect of earth worm meal on nutrient di-
gestibility, nutrient absorption, and cholesterol content
in the blood and meat of the birds.
REFERENCES
Balamurugan, M., K. Parthasarathi, E. L. Cooper & L.
S. Ranganathan, 2007. Earthworm paste (Lampito
mauritii, Kinberg) alters inflammatory, oxidative,
haematological and serum biochemical indices of
inflamed rat. European Review for Medical and
Pharmacological Sciences. 11(1):77–90.
Balamurugan, M., K. Parthasarathi, L. S. Ranganathan,
& E. L. Cooper, 2008. Hypothetical mode of action
of earthworm extract with hepatoprotective and
antioxidant properties. J. Zhejiang Univ. Sci. B. 9(2):
141–147.
Chumaidi, 2005. Peluang cacing tanah sebagai substitusi
tepung ikan untuk pakan induk ikan. Warta Penelitian
Perikanan Indonesia. 11 : 15-18.
Cooper, E. L. 2005. CAM, eCAM, bioprospecting: The 21st
century pyramid. Evidence-based Complementary
and Alternative Medicine, 2(2):125–127.
Damayanti, E., A. Sofyan, H. Julendra & T. Untari. 2009.
The use of earthworm meal (Lumbricus rubellus)
as anti-pullorum agent in feed additive of broiler
chicken. Journal (JITV). 14(2). Abstract.
Indon. J. Nutr. and Feed Sci.
Ismail, S. A., K. Pulandiran, R. Yegnanarayan. 1992. Anti-
inflammatory activity of earthworm extracts. Soil
Biology Biochemistry, 24(12):1253–1254.
Mambo. 2006. Sabut Kelapa dan Ampas Tahu sebagai
Media dan Pakan Cacing Tanah (Lumbricus rubellus).
http://maluku.litbang.deptan.go.id. [2 July 2009]
Mudjiman, A. 2004. Makanan Ikan. Edisi revisi. Penebar
Swadaya, Jakarta.
Prakash, M., M. Balamurugan, K. Parthasarathi, G.
Gunasekaran, E.L. Cooper, & L.S. Ranganathan,
2007. Anti-ulceral and anti-oxidative properties of
“earthworm paste” of Lampito mauritii (Kinberg) on
Rattus Norvegicus. European Review for Medical and
Pharmacological Sciences, 11(1):9–15.
Purwakusuma, W. 2007. Lumbricus rubellus sebagai
Alternatif Pakan Ikan. Available at : http://O-fish.
com. [2 July 2009].
Rahman, 2008. Pembuatan Pakan Ikan Alternatif dari Bahan
Cacing Tanah. Available at : http://forum.O-fish.com/
viewtopic.php?f=30&t=21896. [2 July 2009].
Resnawati, H., 2004. Bobot potongan karkas dan lemak
abdomen ayam ras pedaging yang yang diberi
ransum mengandung tepung cacing tanah (Lumbricus
rubellus). Prosiding Seminar Nasional Teknologi
Peternakan dan Veteriner. Balai Penelitian Ternak
Ciawi. 10 (1) : 110 (Abstract).
Resnawati, H., 2008. Respon ayam pedaging terhadap
ransum yang mengandung tepung cacing tanah
(Lumbricus rubellus). Available at : http://balitnak.
litbang.deptan.go.id/index.php?option=com_content
&task=view&id=124&Itemid=60. [2 July 2009].
Rezeaei, M., H.N. Moghaddam, J.P. Reza & Kermanshahi.
2004. The effect of dietary protein and lysine levels in
broiler performance, carcass and N excretion. Journal
of Poultry Science. 3(2) : 148-152.
Rodriguez, C., V. Beoletti & M. Finola, 1996. Bacteriology
of poultry litter, compost and the earthworm Eisenia
foetida (Oligochaeta, Lumbricidae). Megadrilogica
6:91–95.
Rukmana, R., 1999. Budidaya Cacing Tanah. Kanisius.
Yogyakarta.
Smith, L.W., 2001. The incidence of potentially pathogenic
bacteria in liquor from selected wormfarms. Biol.
Fertil. Soils. 34:215–217.
Soeparno, 1994. Ilmu dan Teknologi Daging. Cetakan
keempat. Gadjah Mada University Press. Yogyakarta.
Waluyo, J., 2008. Purifikasi dan Karakterisasi Protein
Antibakteri dari Cacing Tanah. Available at : http://
adln.lib.unair.ac.id/go.php. [2 July 2009].
Zuprizal, 1993. Pegaruh penggunaan pakan tinggi protein
terhadap penampilan karkas dan perlemakan ayam
pedaging fase akhir. Buletin Peternakan. 17 : 110-
118.
Zuprizal, 2006. Nutrisi Unggas. Fakultas Peternakan.
Universitas Gadjah Mada. Yogyakarta.
January 2011 45
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

INDEX OF AUTHORS
Vol. 2 No. 1

Ariesta, A. A. K., 32 Permana, I. G., 26, 36

Az-zahra, F., 15 Puger, A. W., 10
Cakra, I. G. L. O., 20 Ridla, M., 36
Daisy, L., 6 Santo, E., 6
Damanik, R., 42 Setyawan, I. A., 32
Damry, H. B., 6 Siti, N. W., 20
Despal ., 26 Sumiati., 36
Dono, N. D., 42 Sundu, B., 6
Hanim, C., 15 Supartha, I. W., 10
Hutabarat, I. M. L., 26 Toharmat, T., 36
Jachja, J., 36 Trisnadewi, A. A. A. S., 1
Mastika, I. M., 10 Wibawa, A. A. P. P., 20
Mudita, I. M., 20 Wibowo, A., 42
Murwani, R., 32 Wiranatha, I. W., 10
Mutia, R., 26 Wirawan, I. W., 20
Nahrowi., 36 Yadnya, T. G. B., 1
Pasaribu, J., 42 Yusiati, L. M., 15
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

INDEX OF SUBJECT
Vol. 2 No. 1

abdominal fat, 42 fermented cocoa by product, 10

albumin, 36 fermented coffee pulp, 15
antibiotic, 6 growth performance, 42
bali cattle, 10 hay, 26
beta mannan, 6 in vitro digestibility, 15
broiler, 6 laying hens, 36
broiler chickens, 42 local waste, 20
carcass production, 42 methionine, 36
cellullolytic microbes, 15 metroxylon sagu, 1
chlortetracycline, 32 mungbeans, 32
complete feed, 20 nutrition quality, 20
copra meal, 6 palm kernel meal, 36
corn, 32 physical quality, 1
digestibility, 10 production, 1
duck, 1 ramie leaves, 26
earthworm, 42 rumen liquor, 20
egg, 1, 36 shell, 36
enzyme complex, 20 silage, 26
etawah goat, 26 sorghum, 32
feed conversion, 32 starnox, 1
feed intake, 10
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

INSTRUCTIONS TO AUTHORS OF written in capital letters, bold, and centered.

INDONESIAN JOURNAL OF NUTRITION AND b. Sub-heading is initialized with a capital letter
FEED SCIENCE only the first letter, centered and bold
c. Sub-sub-heading is initialized with a capital
General Aspects letter only the first word, written at the beginning
of the paragraph, bold, and followed by full stop.
1. Only original articles are accepted for publication.
Text on the following line is typed double hit
2. Journal publishes the articles covering research topics
from the sub-sub heading.
in animal nutrition and feed.
3. Articles are written in English. 12. Table
4. The Journal is published in January, May, and a. Table is informative and stands alone.
September. b. Title is typed above the table, numbered using an
Arabic number, initialized by a capital letter, brief
Manuscript File and clear.
c. Horizontal lines separate the head of column, data,
1. Text should be in Microsoft Word format, Table and
and footnote.
Graphs in Microsoft Excel format and Figures in JPEG
d. Data are accompanied with standard deviation
or TIFF format.
(SD), standard error (SE), or coefficient of variation
2. Text should be typed in Times New Roman fonts at
(CV).
12 points, typed double-spaced; Title, Tables, Title of
e. Footnote for statistical analysis is written: “Means
Graphs/Figures, and appendix are typed single-spaced.
in the same column or row with different superscript
3. Manuscripts are prepared in A4 paper with 3 cm margins
differ significantly (P<0.05) or (P<0.01)”.
on all four sides
f. Each abbreviation or symbols is described in a
4. Maximum total number of pages is 20.
footnote.
5. Tables, Graphs, and Figures should be placed separately
from the text. 13. Figure and Graph
6. Page and line numbers should be applied. a. Figure or Graph is informative and stands alone.
7. Content of manuscripts: Title, Name and address b. Title is typed under the Figure or Graph, numbered
of the author(s), Abstract, Introduction, Materials using an Arabic number, initialized by a capital
and Methods, Results and Discussion, Conclusion, letter, brief and clear.
Acknowledgment (if any), References. c. Symbols and description of Figure and Graph should
be defined in title.
Content of the Manuscripts d. Figure and Graph must have good resolution.
1. Title is brief in 14 words of maximum length and e. Standard deviation (SD), standard error (SE), or
initialized with a capitalized letter, clear, and reflects coefficient of variation (CV) must be inserted in
the article content. each Graph.
2. Name of Author(s) are written as for references. f. Bar charts should be made in 2-dimension in the
3. Name of institution(s) indicates full address with simplest shading, not a solid or block shading.
telephone number, and e-mail.
4. Abstract contains objectives, methods, results, and 14. Nomenclature for Latin words is typed in italic. Full
conclusions in one paragraph of maximum 200 words. scientific terminology and the name of chemicals are
5. Key words is typed in italic and no more than 5 (five) used for the first time.
words. 15. Units of measurements are expressed in International
6. Introduction describes the background and objective System (IS).
supported by references. 16. Decimal to separate value is written using dot (.).
7. Materials and Methods should be written clearly and 17. References
fully, especially for materials, procedures, experimental a. References consist of at least 80% of the last 10
design, and data analysis. years publication in journal.
8. Results and Discussions contains results, discussion b. Dissertation, thesis, patent document, book are
regarding the problems investigated, and supporting allowed to include up to 20% of the total references.
references. Proceeding is not allowed.
9. Conclusion reflects the experimental results. c. Electronic publications are only allowed to use, if
10. Acknowledgement expresses thanks to person and those are published by a competent institution, and
institution. put the accessed date after the last sentence.
11. Heading d. References are listed alphabetically by the
a. Headings (ABSTRACT, INTRODUCTION, author(s) last name(s) and the year of publication.
MATERIALS AND METHODS, RESULTS Books: author(s) of a book, year, title, name and
AND DISCUSSION, CONCLUSION, place of publisher. Journals: author(s), year, title,
ACKNOWLEDGMENT, REFERENCES) are abbreviation of journal name as suggested by the
INDON. J. NUTR. AND FEED SCI. Vol. 2 No. 1 , January 2011
ISSN 1410-2390

Table 1. Mean of pH, N-NH3 and total VFA concentration, and gas production in vitro as responses to various dietary
treatments.
Dietary treatments
D0 D1 D2 D3 D4
pH 6.97±0.14 6.94±0.09 6.93±0.06 6.92±0.07 6.92±0.07
N-NH3 (mM) 13.52±4.93 12.55±5.56 11.81±4.96 10.81±4.64 13.93±6.02
Total VFA (mM) 82.85±17.39 c
105.54±12.02 ab
114.68±6.99 ab
122.46±7.88 a
98.27±20.30bc
Gas volume (ml) 43.25±11.18 48.75±10.40 49.25±9.84 52.25±08.54 53.50±16.76
Note: Means in the same row with different superscript differ significantly (P<0.05).

700 700

Total length of stolons (cm)

600 600
Total panjang stolon (cm)

500 500
400 400
300 300
200 200

100 100
0 0
0 1 2 3 4 5 6 7 8 9 0 2 4 6 8 10 12 14
Growth Period (week) Growth period (week)

Figure 1. Length of stolons (A) and pattern of stolons growth (B) of B. humidicola grass during 14 weeks growth period: ◊= control,  =
mulch C. odorata, ∆= animal manure, x = combination C. odorata-animal manure, ○ = urea + SP36.

journal, volume and number, and pages. Article in articles as suggested by reviewers.
a book: author(s), year, title of the article, editor(s),
2. Copy right of article(s) published is held by Indonesian
book title, name and place of publisher.
Journal of Nutrition and Feed Science.
e. An example of writing references:
3. Publication of accepted articles is free of charge.
AOAC. 2010. Official Methods of Analysis of
AOAC International, 18th Edition. Assoc. 4. First author get five reprints of accepted articles and
Off. Anal. Chem., Arlington. (Book) one exemplar of the journal.
Biswas, A., M. Ahmed, V. K. Bharti, & S. B.
Singh. 2011. Effect of Antioxidants on
Physiobiochemical and Hematological Address for Correspondence
Parameters in Broiler Chicken at High Editorial Board of Indonesian Journal of Nutrition and Feed
Altitude. Asian-Aust. J. Anim. Sci. Vol. 24: Science
246-249. (Journal) Faculty of Animal Science, Bogor Agricultural University
Jayanegara, A. 2008. Methane reduction effect Jl. Agatis, Kampus IPB Darmaga, Bogor Indonesia 16680
of polyphenol containing plants, simple Telephone: +62 251 8626419
phenols and purified tannins in in vitro rumen Facsimile: +62 251 8626419
fermentation system. Thesis. University of E-mail: jurnal_aini@yahoo.com
Hohenheim, Stuttgart. (Thesis/Disertation)
Peoples, M.B., R.M. Boddey, & D.F. Herridge.
2002. Quantification of Nitrogen Fixation.
In. G. J. Leigh (Ed). Nitrogen Fixation at the
Millennium. Elsevier B.V. Amsterdam. pp.
357-389. (Article in a Book)

Publication and Submission Fee

1. Article(s) submitted for publication are evaluated by
reviewers and editorial board.Author(s) must revise the