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Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
34
Doi: 10.21059/buletinpeternak.v44i2.54713
Growth Performance, Mortality, Relative Organ Weight, Blood Biochemistry, and
Intestinal Microbial of Arbor Acres Broiler Fed Diets Containing Mannan-Riched
Fraction (Mrf) and Probiotic-Enhanced Liquid Acidifier
Danung Nur Adli1, 2 and Osfar Sjofjan3
1Department of Animal Nutrition, University of Brawijaya Kediri, East Java, 64111, Indonesia
2Animal Feed and Nutrition Modelling Research Group (AFENUE), Department of Animal Nutrition and Feed
Technology, Faculty of Animal Science, IPB University, Bogor, 16680, Indonesia
3Department of Animal Nutrition, Faculty of Animal Science, University of Brawijaya, Malang, 65145, Indonesia
Article history
Submitted: 6 March 2020
Accepted: 20 May 2020
* Corresponding author:
Telp. +62 89 880 13588
E-mail:
danungnuradli1994@gmail.com
ABSTRACT
The research purpose was to carry out the effect of mannan-riched fraction
(MRF) and probiotic enhanced water as natural growth promoters (NGPs) on Growth
Performance, mortality, relative organ weight, blood biochemistry, and intestinal
microbial flora. A total of 3000 day old chicks (DOC) Arbor Acres broiler were
randomly allocated to 4 dietary treatments and 4 replications of 187 broilers per cage.
Four treatments used in research were as follows: i) CON, basal diet, ii) basal diet,
CON+ MRF (Actigen™) 80g/100kg/feed , iii) basal diet, CON+ 0.2% drinking water +
2 ml/L Combination feed additive (Acid-Pak 4-way®), and iv) ) basal diet, CON+ MRF
(Actigen™) 80g/100kg/feed+ drinking water 2 ml/L Combination feed additive (Acid-
Pak 4-way®). The results showed that using mannan riched fraction (MRF) (feed) and
combination with probiotic-enhanced liquid acidifier (drinking water) presented
significant difference (P>0.05) on body weight gain at 1-28 days and intestinal
microbial. On the blood biochemistry, the effect of combination began to reduce the
amount of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase
(GPT) at 21 days periods. To sum up, the addition of mannan-riched fraction and
combination with probiotic enhanced liquid acidifier doesn’t impacted on growth
Growth Performance, blood biochemistry, relative organ weight but give significant
effect on intestinal microbial and reduces mortality of broiler.
Keywords: Blood biochemistry, Broiler, Growth Performance, Mannan-rich fractions,
Micro flora
Introduction
Poultry industry serves an essential role in
supporting the availability of cheap animal protein
in Indonesia. This condition is reflected based on
the demand for poultry products nationally. In
2017, broiler production increased approximately
6.82% compared to 6.34% of the population in
2016 (equivalent to 1.6 billion broilers) (Agriculture
Ministry of Indonesia, 2017). However, the poultry
industry mostly involves the use of antibiotics as a
growth promoter. These antibiotics growth
promoters (AGPs) have been in poultry diets as
feed additives for more than 40 years in Europe.
Lately, the use of antibiotic compounds has
decreased, and several European countries has
suppressed the use of antibiotics growth
promoters (AGPs) as non-nutritive feed additives.
Numerous efforts have been undertaken to
develop suitable alternatives to counteract the
anticipated drawbacks associated with the ban of
AGPs. According to the newest regulation,
Indonesia has banned the use of antibiotics in
poultry production (Sjofjan et al., 2020). First the
antibiotics can be toxic to consumers. Secondly,
antibiotics can create resistant microorganisms in
the body of humans or livestock (especially
pathogenic bacteria) (Adli et al., 2018).
The use of antibiotics are still prevalent in
the poultry business. As a result the quality of
meat is depends on the feed and antibiotics use
by farmers as feed additives. Different substances
often referred to as natural growth promoters
(NGPs) are supposed to achieve high consumer
acceptance since they do not usually pose any
risk that will lead to bacterial (Adli et al., 2017).
Although the amount of antibiotic used as
growth promoters is relatively small, it improves
the feed efficiency to help farmers obtain more
enormous profits. Using feed additives is one
method to improve the quality of feed (Jet et al.,
2014) The antibiotics are provided as a growth
promoter; however, they cause bacterial
resistance and residue in the carcass. Alternative
feed additive such as mannan-riched fraction
(MRF) and probiotic enhanced liquid acidifier has
Buletin Peternakan 44 (2): 34-42, May 2020
Bulletin of Animal Science
ISSN-0126-4400/E-ISSN-2407-876X Accredited: 36a/E/KPT/2016
http://buletinpeternakan.fapet.ugm.ac.id/
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
35
been the center of attention for many studies
during the past five years due to its beneficial
effect on feed efficiency. Mannan-riched fraction
belongs to the family of prebiotics using new
techniques such as nutrigenomics. Both prebiotics
and probiotics replaces use of the antibiotics
because they are safer and act as a natural
growth promoter (NGPs) in the broiler (Adli et al.,
2018).
Research in several countries used
prebiotics and probiotics combined for poultry to
enhance their overall Growth Performance and
health. The use of prebiotics combined with
probiotics, acidifier’s even electrolytes in
Indonesia has so far reported to their ability to
maintain health, prevent digestive tract disorder by
utilizing the microbes for balancing and increasing
the population of non-pathogenic bacteria. The
addition of probiotics, prebiotics, and acidifier is
expected to detoxify toxins and their metabolites
to improve absorption of nutrients and reduce
cholesterol level in blood (Sjofjan et al., 2020)
Probiotics are the hot prospects for feed
additives that can be provided to animal in both
solid and liquid forms. The used of prebiotics are
to balance pH, lactic acid bacteria colony, and
decreasing the nutrient of pathogen bacteria that
can survive in the intestines (Jet et al., 2014). The
role of prebiotics has a synergistic effect in which
lactic acid bacteria (LAB) can inhibit the growth of
pathogenic microbes especially Escherichia coli
and Enterococcus sp. The activator of the
prebiotics increases the number of feed intake for
the growth of the internal organ of broiler (Natsir et
al., 2010). Therefore, the purpose of this research
was to investigate the effect of mannan-riched
fraction (MRF) and probiotic enhanced liquid
acidifier on growth Growth Performance, relative
organ weight, blood biochemistry, and intestinal
microbial of broilers.
Materials and Methods
Animals, housing, and experimental design
A total of 3000 (Arbor Acres) broiler with an
initial body weight (BW) of 39.43±1.23 g were
used in a 5-wk trial. Treatments were randomly
assigned to pens within gender using the random
number generator in excel. Pens were assigned in
a randomized complete block design to
compensate for known position effects in the
experimental facility. After randomization of
treatments within gender was completed, average
initial BW for each treatment was checked to
make sure it was equal (4 replicates with 187 per
replication pen). Treatments were as follows: i)
CON, basal diet, ii) basal diet, CON+ MRF
(Actigen™) 80g/100kg/feed, iii) basal diet, CON+
0.2% Drinking water + 2 ml/L Combination feed
additive (Acid-Pak 4-way®), and iv) ) basal diet,
CON+ MRF (Actigen™) 80g/100kg/feed+ Drinking
water 2 ml/L Combination feed additive (Acid-Pak
4-way®). The Actigen™ was obtained from a
commercial company (Alltech Inc., Nicholasville,
Kentucky: USA). A prebiotic was used throughout
this experiment derived from the surface of cell
wall membrane e of a specific strain of yeast -
Saccharomyces cerevisiae 1062. MRF contains a
greater concentration of mannan reactive units
(alpha 1,3 mannan). The feed additive used in this
research was probiotics-enhanced liquid acidifier
consists citric acid, sodium chloride, potassium
chloride, acetic acid, sodium citrate, ethyl vanillin,
zinc sulphate, iron sulphate, magnesium sulphate,
dried Aspergillus niger fermentation extract, and
dried Bacillus subtilis fermentation extract. All
broiler was allowed ad libitum access to feed and
water through a self-feeder and nipple drinker
throughout the experimental period. All broiler was
housed in an environmentally controlled room.
The target room temperature and humidity were
29°C and 64%, respectively. The rice hull-littered
floor pens with height of 3.3 (1.8 x 1.8) m2 per
pen. The lighting program was set at 23 hours
light and one hour darkness.
Growth performance
The arbor acres broilers were individually
weighed at the beginning of the experiment, and
every week thereafter till the end of the
experiment. The gain in body weight (BWG) of
broilers per week was calculated as the difference
between the initial and end weight at a given week
(7 days, 14 days, 21 days, 28 days, and 35 days).
The feed intake was calculated by the difference
between the offered and remained amounts
weekly. The feed intake was calculated after
correction for that used by dead broilers. The feed
gain ratio was calculated by dividing the amount of
feed consumed in a certain period by the gain in
weight at the same period (with consideration of
dead broilers), expressed in the same weight
units. During the experimental period, daily
mortalities were recorded for each group and
mortality rate was calculated (Sjofjan et al., 2019).
Relative organ weight
The 42 broilers from each group, close to
the average live BW, were selected at the days
21, 28, and 35 days broilers will be sacrificed of
the experiment. Broilers were slaughtered by
electrical shock method to obtained organ weight.
In addition, the relative weight of organs, such as
the liver, spleen, pancreas, thymus, bursa, and
spleen of broiler was assessed and its relation to
the live BW of the broiler, in percentage, was
calculated.
Blood biochemistry
Blood samples were collected from the
heart in each group at 21, 28, and 35 days of age.
The Blood non-EDTA tubes obtained from the
samples of broiler and allowed to clot for one
hour, at room temperature, Blood samples was
immediately centrifuged using the cryogenic
centrifuge (Hettich Universal 320R, Germany) for
15min at 3000 rpm. The blood biochemistry and
samples was kept in tubes at−20 °C until
chemically analyzed. At the time of analysis, the
samples were thawed and analyzed for GOT:
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
36
glutamic oxaloacetic trasaminase; GPT : glutamic
pyruvic transaminase; TP: total protein, ALB :
Albumin; GLB : globulin; A/G : albumin/ globulin
ratio; TGL: triglyceride; TCHOL: total cholesterol;
BUN: blood urea nitrogen; GLC: glucose using
Clinical biochemistry Analyzer (CA) (Nicholasville:
Kentucky: USA) (Adli et al., 2019)
Intestinal morphometric
The sample about 6 cm from the middle of
duodenum, jejunum, and ileum were excised and
flushed with ice-cold saline and immediately
placed in combination of liquid of NA2PO4 2%;
NA2H2PO4 2%, 24% Formaldehyde; and 900 ml
reverse osmosis water for morphometric analysis.
The method used were Hematoxylin eosin
staining coloring. The indices of villus height, crypt
depth (µm) and villus height and crypt depth ratio
were measured using computer-aided light
microscope image software m-shot digital image
system with 200x zooming according to method
from (Adli et al., 2019).
Intestinal microbial
Chyme from jejunum and ileum, including
lactic acid bacteria and coliform bacteria were
determined at the end of the experiment. One
gram of the chyme sample from each broiler was
diluted with 9 mL of 10 g/kg peptone broth
(Becton, Dickinson and Co., USA) and
homogenized. Then, 10-fold dilutions of chime
samples were performed (ranging from 10−4 to
10−6) and then cultivated onto MacConkey agar
plates (Difco Laboratories, Detroit, MI, USA) for
the enumeration of coliform bacteria and
lactobacilli agar plates (Medium 222; DSMZ,
Braunschweig, Germany) for the enumeration of
lactic acid bacteria. The MacConkey agar plates
were incubated for 24 hours at 37°C. The
lactobacilli agar plates were then incubated for 48
hours at 37°C under anaerobic conditions. The
lactic acid bacteria and coliform bacteria colonies
were counted immediately after removal from the
incubator. Concentration of microflora was finally
expressed as log10 colony-forming units per gram
of chime (Adli et al., 2019).
Data analyses
The statistical analyses were performed
Analysis of variance (anova) tests were used for
analyses of variance accompanied by Duncan’s
multiple range test to detect the differences
between the treatments with help software using
SAS (version 9.4, SAS University Ver. Inc.,). The
results are presented as standard error mean
(SEM) among group treatment. Probability values
less than 0.05 (p<0.05) was considered significant
(Widiyawati et al., 2020).
Results and Discussion
Growth performance
According to the Table 2 shows that the
growth performance, diets containing MRF and
probiotics-enhanced liquid acidifier level was
presented no significant difference (P<0.05) on
body weight, feed intake, and feed/gain but on
body weight gain at 1-28 days for all treatment
(T1, T2, and T3) increased compared with control
(1138.73, 1128.63, 1192.09 vs. 1001.12 g/broiler;
P>0.05). The treatment (T3) presented greater
than all treatment since it combination mix both
give in drinking water and experimental diet may
stimulation the increasing of body weight gain.
Table 1. Experimental diet
Feed nutrient (%)
Starter (1-21 days)
Finisher (22-35days)
Yellow corn
57.11
69.66
Dehulled soybean meal
36.53
26.65
L-Lysine
0.10
0.10
DL-methionine
0.55
0.55
Dicalcium phosphate
1.67
1.55
Limestone
1.13
1.02
Salt
0.30
0.30
Soy oil
2.81
0.06
Vitamin premix*
0.05
0.05
Mineral premix**
0.05
0.05
Choline
0.10
0.10
100
100
Dry matter (%)
87.00
87.00
ME (Kcal/kg)
3050
3150
Ash (%)
9.00
9.00
Crude protein (%)
22.00
18.00
Fat (%)
6.00
6.00
Crude fibre (%)
3.00
2.50
Ca (%)
1.00
0.95
P (%)
0.70
0.75
Copper (ppm)
30
50
Zinc (ppm)
120
120
*vitamin premix (per kg of diet); vitamin A 12,500 IU; Vitamin D3, 2,500; Vitamin E 20 IU; Vitamin K3 2.5 Mg; Vitamin B1 2Mg; Vitamin B2
5 Mg; Vitamin B6 3Mg; Vitamin B12 0.012 Mg; Niacin 35 Mg; Pathonenic acid 12Mg; Folic Acid 1Mg;
**Mineral premix (Per kg of diet); Fe 70 mg, Zn, 90 mg; CU, 10 mg; Mn, 80 mg
The treatment use four treatments, and four replicates. The treatments were:
T0: Basal diet + without feed additive (control); T1: Basal diet + MRF (Actigen™) 80g; T2 : Drinking water + 2 ml/L Combination feed
additive (Acid-Pak 4-way®); and T3: Basal diet + MRF (Actigen™) 80g + Drinking water 2 ml/L Combination feed additive (Acid-Pak 4-
way®).
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
37
Table 2. Effect of average group of MRF and probiotics-enhanced liquid acidifier on the body weight and body weight gain of broiler
Treatments1
Day
T0
T1
T2
T3
SEM
— — — —Body weight, g/broiler— — — —
1
43.72
43.65
43.65
43.78
0.30
21
718.40
713.70
704.39
715.92
37.20
28
1212.20
1245.20
1220.20
1235.70
37.60
35
1842.55
1889.30
1842.00
1775.25
70.60
— — — —Body weight gain, g/broiler— — — —
1-21
674.64
670.05
671.98
668.98
42.55
1-28
1001.12b
1138.73a
1128.63a
1192.09a
47.70
21-35
1525.10
1537.10
1574.70
1486.90
65.20
1-35
1856.90
1842.70
1878.40
1786.60
167.20
a,b Mean values not sharing the same superscripts in a row differ significantly (P < 0.05);
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®).
The combination of MRF and probiotic helps the
treatment (T3) in blood and increase body weight
gain of broilers. Compared with Brennan et al.
(2014) stated the used of MRF give significant
difference (P<0.05) on body weight gain at 21 d
and 35 d compared than control (877 g (MRF 400
g/ton (21 d); 50 g/tonne (35d)) vs 819 g control)
due it combination both of mannan and probiotic.
The result according to the table 2 got additional
statement from Brennan et al. (2014) stated that
the result of performance broiler were according
due to factor rearing condition, the broiler will
increase the body weight gain when the
environment (bedding are clean) (Brennan et al.,
2014). The result continued to the Table 3
showing the feed intake was no significant
difference (P>0.05) at whole periods. The feed
intake result shown on the table 3 were no
significant difference at 1-28 days treatment (T1)
better than control (2018.40 vs 1874.60 g/broiler),
21-35 days (T1, and T2) give best result
compared than control (2414.40, and 2273.40 vs
2264.70 g/broiler) and followed by result at 1-35
days (T1, and T2) better than control (T0)
(2810.80, and 2690.10 vs 2668.60 g/broiler). The
feed intake increased may due correlating with
body weight and body weight gain, when both of
thus variable growth increase the feed intake will
also increase. The increase may due to rearing
condition, the way of feed give ad-libitum made
broiler eat more. According to Brennan et al.,
(2018) stated feed intake at 21 d and 35 d
compared than control (1.33 (MRF 400 g/ton (21
d); 1.49/ton (35d)) vs 1.23 control) due its rearing
condition factor example the way and quality of
feed given. The mortality result on table 3 showed
the used of MRF combination with probiotic liquid
acidifier on treatment (T2 and T3) give no
significant differences (P>0.05) reduces to 1.31%
compared to control 3.94%. The mortality were
also parameters observed on the research to
combine the relevance it with treatment reduces
percentage or not. The result may include the
composition in the feed additive can prevent
factors dead factors such as coccidian during the
research. In the composition the Saccharomyces
cerevisiae 1062, citric acid, sodium chloride,
potassium chloride, acetic acid, sodium citrate,
ethyl vanillin, zinc sulphate, iron sulphate,
magnesium sulphate, dried Aspergillus niger
fermentation extract, and dried Bacillus subtilis
helps to stimulate immune modulation of broilers,
Contrary to these findings, it was reported from
study Biswas et al. (2018) cannot help to reduce
mortality in male broilers at 0-21 d and 0-42 d
(3.38 vs 0.28 (control)) and (4.51 vs 1.69 (control))
because the experimental the broiler reared under
stocking density stress (43 kg live weight per m2
floor space) (Biswas et al., 2018).
Relative organ weight (%)
The result was not significantly difference
(P>0.05) for liver, spleen, bursa of fabricius,
thymus, and pancreas of organ weight at 21, 28,
and 35 days. However, the immune organ such as
thymus are better on the (T1, T2, and T3)
compared to control (T0) (table 4) at 21 days for
all treatment (0.38, 0.31, and 0.33 vs. 0.30)
although, it not significant difference (P>0.05) on
statistics.
Table 3. Effect of MRF and probiotics-enhanced liquid acidifier on the feed intake, feed/gain, and mortality of broiler
Treatments1
Day
T0
T1
T2
T3
SEM
— — — —Feed intake, g/broiler— — — —
1-21
909.80
979.10
855.40
807.30
21.06
1-28
1874.60
2018.40
1822.90
1727.80
53.70
21-35
2264.70
2414.40
2273.40
2102.00
70.00
1-35
2668.60
2810.80
2690.10
2499.00
76.80
— — — —Feed/gain, g/broiler— — — —
1-21
1.33
1.49
1.22
1.22
0.09
1-28
1.73
1.72
1.54
1.44
0.23
21-35
1.52
1.58
1.54
1.49
0.37
1-35
1.48
1.54
1.50
1.44
0.14
— — — —Mortality, (%)— — — —
1-35
3.94
5.26
1.31
1.31
4.36
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
38
The bursa of fabricius in 35 days presented
not significant difference (P>0.05) (Table 5) (0.12,
0.12, and 0.15 vs. 0.12). In the experiment, the
stabile weight of bursa of fabricius could have an
initial positive signal for the development of broiler
immune systems. However, as broiler age more,
the bursa of fabricius will disappear according to
the maturation of the immune system of the
broiler. The weight of bursa fabricius (percentage
of live weight) using MOS 0.1% and 0.2% are not
significance difference (P>0.05) (0.28 and 0.32 vs
0.29 (control)) at finisher periods of broiler (Biswas
et al., 2018). The weight of liver, spleen and
pancreas also are not significant difference
(P>0.05) (Biswas et al., 2018). The exact
explanation may due to prebiotic helps protect
proliferating immature bursal B cells and thymic T
lymphocytes from oxidative stress (Biswas et al.,
2018). The studies from the used MOS in relative
organ weight of male broiler at level 1 g/kg were
not significant influenced (P>0.05) (0.19 (MOS)
vs. 0.20) it because the activities from MOS
sometimes not occur (Bozkurd et al., 2009). In
addition, the bursa of fabricius is an organ of the
immune system and is responsible for maturation
of B-lymphocytes (Biswas et al., 2018).
Intestinal microbial
Based on table 5 the used MRF and
probiotic-enhanced liquid acidifier as a feed
additive on intestinal microbial were not significant
different (P>0.05) on the Lactobacillus and
Coliform at 21, 28, and 35 days both jejunum and
ileum. At 28 days the Coliform in the ileum parts
the trends were decreasing even though, were not
significant different (P>0.05) (2.74 (T1), 3.03 (T2),
3.11 (T3) vs. 3.23 (T0) Log cfu/g, DM). However,
the Lactobacillus decreasing at small amount at
ileum parts (2.33 (T1), 3.81(T2), and 3.40(T3) vs
4.36 (T0) Log cfu/g, DM). At 35 days (table 7) the
Lactobacillus were not significant difference
(P>0.05) for T1,T2,T3 in line but still better than
control (T0) (4.78, 4.56, 4.72 vs 4.32 Log cfu/g,
DM). At 28 days, the positive microbial population
may starts to stabilize indicated the treatment may
help the microbial stable on bacterial fermentation,
but at 35 d the condition began unstable at
several cases.
Table 4. Effect of average group MRF and probiotics-enhanced liquid acidifier on the relative organ weight of broiler
Treatments1
Day
Item(g)
T0
T1
T2
T3
SEM
— — ( Organ weight/body weight ) x 100 — —
21
Liver
3.31
3.16
2.62
3.24
0.41
Spleen
0.10
0.10
0.12
0.12
0.02
Bursa
0.22
0.21
0.32
0.20
0.09
Thymus
0.30
0.38
0.31
0.33
0.11
Pancreas
0.39
0.46
0.43
0.45
0.07
28
Liver
2.85
2.83
2.83
2.18
0.42
Spleen
0.17
0.11
0.17
0.09
0.04
Bursa
0.20
0.17
0.17
0.14
0.05
Thymus
0.28
0.28
0.31
0.26
0.10
Pancreas
0.32
0.30
0.31
0.29
0.05
35
Liver
1.86
1.89
2.02
1.83
0.15
Spleen
0.08
0.11
0.12
0.12
0.02
Bursa
0.12
0.12
0.12
0.15
0.03
Thymus
0.32
0.29
0.33
0.26
0.10
Pancreas
0.20
0.21
0.20
0.22
0.02
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Table 5. Effect of average group MRF and probiotics-enhanced liquid acidifier on the intestinal microbial of broiler
Treatments1
Bacterial
Day
T0
T1
T2
T3
SEM
Jejunum
21 — — — — Log cfu/g, DM — — — —
Lactobacillus
2.47
2.72
1.83
2.66
0.87
Coliforms
2.44
1.51
1.55
3.50
1.73
Ileum
Lactobacillus
2.34
2.83
1.73
2.08
0.79
Coliforms
2.62
3.33
3.52
2.43
0.09
Jejunum
28
— — — — Log cfu/g, DM — — — —
Lactobacillus
3.17
3.35
2.98
3.44
0.63
Coliforms
2.85
3.04
3.21
3.00
1.73
Ileum
Lactobacillus
4.36
2.33
3.81
3.40
1.09
Coliforms
3.23
2.74
3.03
3.11
1.59
— — — — Log cfu/g, DM — — — —
Jejunum
35
Lactobacillus
4.56
4.44
3.96
3.57
1.25
Coliforms
2.34
2.12
2.77
3.32
1.56
Ileum
Lactobacillus
4.32
4.78
4.56
4.72
0.35
Coliforms
2.56
2.78
2.54
2.31
1.13
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
39
The result showed on Table 6 the used
MRF and probiotic-enhanced liquid acidifier as a
feed additive on intestinal properties were
significant different (P<0.05) on the jejunum part.
At 21 days, (Table 8) Villus height were significant
difference (P<0.05) the treatment (T1: MRF
80g/100kg;T2: Drinking water + 2 ml/L (Acid-Pak
4-way®),T3: MRF 80g/100kg + Drinking water 2
ml/L (Acid-Pak 4-way®)) help to increase surface
area of intestinal compared to control (593.00
(T1), 597.50 (T2), and 569.50 (T3) vs 442.75 (T0)
µm). However, based on table 8 VH/CD whole are
not significant difference (P>0.05). The result
may be correlated with the treatment of probiotics,
and prebiotics that had helped to increase the
surface area of the morphology of small intestine
(jejunum). Compared Brennan et al., (2014) The
used of MRF 7 d to 21 (400 g/tonne); 21 to 42 d
(200 g/tonne) better than control (1267.3 vs 796.6)
increase villus height of broiler (P<0.05), villus
height in jejunum tissue increase is a positive
indicator of intestinal health and increased of
absorptive area F. The level addition of a probiotic
to broiler increased the villus height leading to
increased intestinal surface area and therefore to
increased digestion and absorption of nutrients in
the basal diet (Brennan et al., 2014). The result
(table 8) at 35 days the used of MRF and probiotic
liquid acidifier give significant difference (P<0.05)
on villus height and crypth depth for the treatment
(T1: MRF 80g/100kg;T2: Drinking water + 2 ml/L
(Acid-Pak 4-way®),T3: MRF 80g/100kg+ Drinking
water 2 ml/L (Acid-Pak 4-way®)).
a) b)
a) b)
c) d)
Figure 1. Representative condition of a) 21 days intestinal characteristic condition;
b) 28 days intestinal characteristic condition; c) 35 days intestinal characteristic condition;
d) VH/CD at 35 days intestinal characteristic condition (T3).
Note: (a shallow crypt is positive factors for the development
of an immune status and efficient for the small intestine).
Table 6. Effect of average group MRF and probiotics-enhanced liquid acidifier on the intestinal microbial at 21, 28, and 35 days of age
Treatments1
Jejunum
T0
T1
T2
T3
SEM
21 days
— — — — µm— — — —
VillusHeight
442.75b
593.00a
597.50a
569.50a
57.57
Crypt depth
83.50
134.75
143.75
121.75
46.50
VH/CD
5.30
4.40
4.15
4.67
6.74
28 days
VillusHeight
378.00c
617.00ab
666.00a
508.50bc
63.72
Crypt depth
105.00b
140.00ab
170.00ab
136.25ab
25.01
VH/CD
3.60
4.40
3.91
3.73
1.34
35 days
VillusHeight
598.50b
713.75a
695.50a
718.50a
49.34
Crypt depth
112.50b
148.75ab
149.00ab
156.50a
25.17
VH/CD
5.32
4.79
3.99
4.59
3.45
a,b Mean values not sharing the same superscripts in a row differ significantly (P<0.05);
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
40
The treatment (T3) give the greater
compared to all treatment and control (718.50 vs
713.75 (T1), 695.50 (T2), and 598.50 (control)
µm; villus height). It may the combination both of
MRF and probiotic liquid enhanced acidifier helps
to increase surface are at final days. While, the
villus height increase it correlation with crypt depth
also increase (148.75 (T1), 149.00 (T2), and
156.50 (T3) vs 112.50 (control) µm). The MRF
given at uniform dose (200, 400 mg/kg) are not
significant different in crypt depth ratio (42d) (Di
Giola and Biavati, 2018). A larger area of cryth
depth may positive and faster growing in tissue for
helps maintenance energy requirements (Spring
et al., 2015). Based on the figure 1, a shallow
crypt is positive factors for the development of an
immune status and efficient for the small intestine.
With a lower renewal rate, the cells in the
intestinal become mature and allowing more
efficient digestive enzyme production and
absorption (Van Nevel et al., 2005).
Blood biochemistry
Based on Table 7 and 8 the used MRF and
probiotic-enhanced liquid acidifier as a feed
additive on serum blood biochemistry were not
significantly different (P>0.05) but the results on
Glutamic oxaloacetic transaminase (GOT) at 21
days began to reduce. The treatment better
compared to control (206.25 (T1), 208.25 (T2),
and 228.00 (T3) vs. 238.50 U/L). The criteria for
GOT were < 40 U/L for broiler. Continued to GPT
the treatment were began trends were reduced at
28 days compared to control (table 6) (1.75 (T1),
2.00 (T2), 2.00 (T3) vs 2.25 U/L) even though,
were not significantly different (P>0.05). The
indicator normal for broiler were at < 41 U/L. The
amount of GOT and GPT were unstable at 35
days (Table 7). The GOT treatment better
compared to control (300.80 (T1), 265.80 (T2),
and 343.30 (T3) vs. 329.50 U/L) and GPT were
(3.00 (T1), 1.75 (T2), and 2.00 (T3) vs. 2.75 U/L).
The result may due to the treatment cannot help
to reduce the amount of GPT and GOT. The GPT
and GOT were the indicator in the liver that the
treatment cause negative effect or not. The dietary
treatments did not have significant effects on the
activities of GOT and GPT because the ability
from probiotics does not always occur it depends
on the optimum dose, frequency, and duration of
treatments (An et al., 2008).
Based on Table 7 and 8 the used MRF and
probiotic-enhanced liquid acidifier as a feed
additive on serum blood biochemistry were not
significantly different (P>0.05) but the results on
total protein (TP) at 21 days still stabile. The
treatment better compared to control (2.57 (T1),
2.52 (T2), and 2.55 (T3) vs. 2.57 g/dL (21 days))
and (2.97 (T1), 2.97 (T2), 2.92 (T3) vs 2.95 g/dL
(35 days)). The criteria for TP were < 2.55 g/dL for
broiler. Continued to albumin (ALB) the treatment
were cannot help to reduce trends at 21-35 days
Table 7. Effect of average group MRF and probiotics-enhanced liquid acidifier on the serum blood biochemistry of broiler at 21 days of
age
Treatments1
Item3
T0
T1
T2
T3
SEM
GOT (U/L)
238.50
206.25
208.25
228.00
21.97
GPT(U/L)
2.00
1.50
2.25
2.75
0.61
TP (g/dL)
2.57
2.57
2.52
2.55
0.17
ALB (g/dL)
1.05
1.02
1.05
1.12
0.15
GLB (g/dL)
1.55
1.57
1.37
1.62
0.12
(A/G)
0.70
0.67
0.70
0.65
0.09
TGL(mg/dL)
140.25
141.75
142.00
144.25
97.65
TCHOL(mg/dL)
130.25
131.75
118.00
122.25
17.70
BUN(mg/dL)
1.27
1.05
1.06
1.27
0.25
GLC(mg/dL)
255.50
278.75
238.50
234.75
36.83
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Table 8. Effect of average group MRF and probiotics-enhanced liquid acidifier on the blood biochemistry broiler at 35 days of age
Treatments1
Item3
T0
T1
T2
T3
SEM
GOT (U/L)
329.50
300.80
265.80
343.30
115.20
GPT(U/L)
3.00
1.75
2.00
2.75
0.97
TP (g/dL)
2.97
2.97
2.92
2.95
0.29
ALB (g/dL)
1.27
1.27
1.17
1.22
0.16
GLB (g/dL)
1.70
1.70
1.77
1.70
0.18
(A/G)
0.77
0.77
0.67
0.72
0.11
TGL(mg/dL)
32.25
30.25
30.50
29.00
5.66
TCHOL(mg/dL)
128.50
128.75
114.50
123.50
12.38
BUN(mg/dL)
0.97
0.47
0.62
0.47
0.32
GLC(mg/dL)
251.00
205.00
213.75
227.00
29.10
1T0= control; T1= MRF 80g/100kg; T2=Drinking water + 2 ml/L (Acid-Pak 4-way®); T3=MRF 80g/100kg+ Drinking water 2 ml/L (Acid-
Pak 4-way®)
Danung Nur Adli and Osfar Sjofjan Growth Performance, Mortality, Relative Organ Weight
41
compared to control (table 6) (1.05 (T1), 1.02 (T2),
1.05 (T3) vs 1.12 g/dL (21 days)) and (1.27 (T1),
1.27 (T2), 1.17 (T3) vs 1.22 g/dL (35 days)) even
though, were not significantly different (P>0.05).
The indicator normal for broiler were at <
1.00 g/dL. The amount of GLB and TGL were
unstable at 35 days (table 7). The GLB treatment
better compared to control (1.70 (T1), 1.70 (T2),
and 1.77 (T3) vs. 1.70 (T0) g/dL) and TGL were
(30.25 (T1), 30.50 (T2), and 29.00 (T3) vs. 32.25
mg/dL). High triglycerides may contribute to
hardening of the arteries or thickening of the
artery walls (arteriosclerosis) — which increases
the risk of stroke, heart attack and heart disease.
The result may due to the treatment cannot help
to reduce the amount of TP, ALB and GLB. The
ALB and GLB were the indicator in the liver that
the treatment cause negative effect or not. The
dietary treatments did not have significant effects
on the activities of ALB and TGL because the
levels in treatment doesn’t helps, which contains
antioxidants (An et al., 2008).
Based on Table 7 and 8 the used MRF and
probiotic-enhanced liquid acidifier as a feed
additive on serum blood biochemistry were not
significantly different (P>0.05) but the results on
total cholesterol (TCHOL), blood urea nitrogen
(BUN), and glucose (GLC) at 21 days began to
reduce. The treatment (T2 and T3) better
compared to control (206.25 (118.00 (T2), and
122.25 (T3) vs. 130.25 mg/dL). The criteria for
GOT were < 115 mg/L for broiler. Continued to
BUN the treatment were began trends were
reduced at 35 days compared to control (table 6)
(0.47 (T1), 0.62 (T2), 0.47 (T3) vs 0.97 U/L) even
though, were not significantly different (P>0.05).
The indicator normal for broiler were at <
0.47 U/L. The amount of GLC were unstable at 21
days (table 7). The GLC treatment (T2 and T3)
better compared to control (238.50 (T2), and
234.75 (T3) vs. 255.50 mg/dL). When our glucose
levels are optimal, it often goes unnoticed. But
when they stray from recommended boundaries,
while glucose notice the unhealthy effect it has on
normal functioning. The dietary treatments did not
have significant effects on the activities of TCHOL,
BUN, GLC because the ability from probiotics
does not always occur it depends on the optimum
dose, frequency, and duration of treatments (An et
al., 2008).
Conclusion
To sum up, the addition of mannan-riched
fraction and combination with probiotic enhanced
liquid acidifier doesn’t impacted on growth Growth
Performance, blood biochemistry, relative organ
weight but give significant effect on intestinal
microbial and reduces mortality of broiler.
Acknowledgment
The authors are grateful to the corporation
which has facilitated Actigen™ (Alltech Inc.,
Nicholasville, Kentucky: USA) for the learning
process in Taiwan.
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