Effects of probiotic, prebiotic and synbiotic intake on blood enzymes and performance of Japanese quails (Coturnix Japonica)

Abstract

The present study was conducted to evaluate the influence of probiotic, prebiotic and synbiotic on performance values and blood enzymes of Japanese quails. One-day-old Japanese quail chicks (192), mean body weight 7.78 g, were randomly assigned in 4 treatments and 4 replicates. The experimental diets consisted of a basal diet without additive (control), 0.2 g/kg probiotic (P), 1.6 g/kg prebiotic (F) and 1.0 g/kg synbiotic (B) added to the basal diet. Birds fed synbiotic showed a significant elevation in body weight compared to other groups. Feed intake of birds fed synbiotic and prebiotic were higher than control and probiotic fed groups. Birds fed synbiotic exhibited a better feed conversion ratio (3.08) compared to probiotic fed (3.19) and control groups (3.14). The males fed additives showed decrease in liver weight. Females fed prebiotic showed decrease in liver weight compared to control and synbiotic additive groups. The relative weight of heart was decreased in the males fed prebiotic compared to synbiotic fed and control groups. The females fed probiotic showed increase in heart weight. The activity of ALP in males elevated by synbiotic consumption. ALP activity decreased in females fed additives. ALT activity was depressed in males fed probiotic or synbiotic. AST activity in males fed prebiotic elevated. In both gender CPK activity was higher in prebiotic feeding group. Results indicated that using synbiotics has positive effects on performance and normal activity of enzymes and prebiotic has positive effects on performance and reduction weigh of heart and liver in Japanese quails.

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Present address: 1,4,5 Faculty of Veterinary Medicine (e mail:
Babazadeh@yahoo.com; Jafargholipour@yahoo.com;
sinavahdatpour@yahoo.com); 3 (e-mail: nikpiran@yahoo.com)
Department of Clinical Science; 2 (e mail: tohid_vahdatpour
@yahoo.com) Department of Animal Science, Islamic Azad
University, Shabestar Branch, Shabestar, Iran.
Indian Journal of Animal Sciences 81 (8): 870–874, August 2011
Effects of probiotic, prebiotic and synbiotic intake on blood enzymes and
performance of Japanese quails (Coturnix japonica)
D BABAZADEH1, T VAHDATPOUR2, H NIKPIRAN3, M A JAFARGHOLIPOUR4 and S VAHDATPOUR5
Islamic Azad University, Tabriz, Iran
Received: 19 March 2011; Accepted: 16 April 2011
ABSTRACT
The present study was conducted to evaluate the influence of probiotic, prebiotic and synbiotic on performance
values and blood enzymes of Japanese quails. One-day-old Japanese quail chicks (192), mean body weight 7.78 g, were
randomly assigned in 4 treatments and 4 replicates. The experimental diets consisted of a basal diet without additive
(control), 0.2 g/kg probiotic (P), 1.6 g/kg prebiotic (F) and 1.0 g/kg synbiotic (B) added to the basal diet.
Birds fed synbiotic showed a significant elevation in body weight compared to other groups. Feed intake of birds fed
synbiotic and prebiotic were higher than control and probiotic fed groups. Birds fed synbiotic exhibited a better feed
conversion ratio (3.08) compared to probiotic fed (3.19) and control groups (3.14).
The males fed additives showed decrease in liver weight. Females fed prebiotic showed decrease in liver weight
compared to control and synbiotic additive groups. The relative weight of heart was decreased in the males fed prebiotic
compared to synbiotic fed and control groups. The females fed probiotic showed increase in heart weight. The activity
of ALP in males elevated by synbiotic consumption. ALP activity decreased in females fed additives. ALT activity was
depressed in males fed probiotic or synbiotic. AST activity in males fed prebiotic elevated. In both gender CPK activity
was higher in prebiotic feeding group.
Results indicated that using synbiotics has positive effects on performance and normal activity of enzymes and
prebiotic has positive effects on performance and reduction weigh of heart and liver in Japanese quails.
Key words: Enzyme, Feed additives, Japanese quail, Performance
The continued feeding of antibiotics at sub-therapeutic
levels to poultry has created concerns about the extent to
which usage increases the possibilities of antibiotic residue,
the development of drug-resistant bacteria, and a reduction
in the ability to cure these bacterial diseases in humans
(Donoghue Dan 2003). Increased awareness of the potential
problems associated with the use of antibiotics has stimulated
research efforts to identify alternatives to their use as feed
additives. In present study a probiotic, a prebiotic and a
synbiotic were chosen as feed additives.
Probiotic, live microorganisms, confer a health benefit
on the host (FAO/WHO 2002). Prebiotics are nondigestible
food ingredients that beneficially affect the host by selectively
stimulating the growth and/or activity of one or a limited
number of bacteria in the colon (Gibson and Roberfroid
1995). The prebiotic approach does not have a long history
of use in broiler chickens (Yang et al. 2009).
The goal of present study is indicating the effects of
commercially procured probiotic, prebiotic and synbiotic on
serum enzymes and growth performance values in male and
female quails.
MATERIALS AND METHODS
Experimental design and housing: Day-old Japanese quail
chicks (192), mean body weight 7.78±0.39 g, that were
randomly assigned in 16 pens with 12 birds (6 males and 6
females) per pen and each bird occupied 0.015 m- 2 of wiry
floor space. The pens were randomized with respect to feed
additives. Temperature was maintained at 35°C for the first
5 days and then gradually reduced according to normal
management practices until a temperature of 22°C was
achieved. Continuous lighting was maintained in all
experimental period (2.5 watt/m 2).
Treatments and additives: The experimental design was
a completely randomized design (CRD), with 4 treatments
and 4 replicates for each treatment. Nutrients compositions
of diets for quails at 1 to 42 days old were based on the
National Research Council (NRC 1994) recommendations

August 2011] EFFECTS OF ADDITIVES INTAKE ON JAPANESE QUAILS 871
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(Table 1). Treatment groups followed of: (1) basal diet
without additive; (2) basal diet plus a multi-strain probiotic
in dry white powder form (2×109 cfu/g: containing
Streptococcus salivarius sub sp. Thermophilus, Lactobacillus
(L) delbruckii sub sp. bulgaricus, L. acidophilus, L.
plantarum, L. rhamnosus, Bifidobacterium bifidum,
Enterococcus faecium, Candida pintoloppesii, and
Asperigillus oryzae) at level of 0.2 g/kg; (3) basal diet plus a
prebiotic (Aspergillus meal) at level of 1.6 g/kg; (4) basal
diet plus a synbiotic (a combination of Enterococcus faecium
as probiotic strain, oligosaccharides as prebiotic, phytogenic
substances and cell wall fragments) at level of 1 g/kg.
Balanced diets were given ad lib. for all treatments at 1 to 42
days old.
Blood sampling and measurements: Feed intake (FI) of
each experimental unit (each cage) was recorded. At the end
of experimental period (42 d) the total body weight (BW) of
birds in each cage was measured and then feed conversation
ratio (FCR) was calculated. Before slaughtering the final BW
of sample bird and after that weight of selected organs
including liver and heart were recorded individually and
presented as a percentage of live body weight. At 42 day’s
of age in fasting state, blood samples were collected by
cervical cutting of 2 birds (1 male and 1 female) per pen
(N,8) and rapidly were centrifuged at 5000 rpm during 5
min and then sera was analysed by using commercial kits
for aspartate aminotransferase (AST), alkaline phosphatase
(ALP), alanine transaminase (ALT), gamma glutamyl
transpeptidase (GGT), lactate dehydrogenase (LDH) and
creatine phosphokinase (CPK) in auto analyzer (IFCC 2011).
Statistical analysis: The data of experiment were analyzed
by an analysis of variance (ANOVA) using the general linear
model (GLM) procedure of SAS (2001) and means were
compared by Duncan‘s multiple Range test at P<0.05 level
(Duncan 1955).
RESULS AND DISCUTION
Studies showed some favourable responses for FI and FCR
by dietary additives. Inborr (2000) reported that prebiotics
(mainly oligosaccharides), probiotics and antibiotics
markedly improved the general health status and FCR of the
poultry. Similarly, some workers have reported increased
growth and improved FCR as a consequence of fructo-
oligosaccharide (FOS) inclusion in broiler diets (Fukata et
al. 1999, Wut et al. 1999). The effects of 3 different types of
feed additives that were added to control basal diet on growth
performance values are presented in Table 2.
In present study, FI in birds fed synbiotic and birds fed
prebiotic was higher (P<0.05) than birds of control group
and birds fed probiotic. Present study indicates that prebiotic
and synbiotic could improve feed intake compared to control
group. Birds fed synbiotic additive have better FCR compared
to probiotic additive and control groups (P<0.05). FCR was
intermediate state for birds fed prebiotic. BW which is an
important indicator of production and lighter birds on the
average produce the lowest meat and egg mass, because this
class would include mainly more unhealthy birds than the
heavier birds (Singh and Nordskog 1982). In present study,
BW in birds fed synbiotic was higher than probiotic fed and
Table 1. Ingredient and calculated analysis of basal diet
Ingredients Ration (%)
Yellow corn 53.00
Soybean meal, 44%CP 37.00
Fish meal, 60%CP 5.50
Vegetable oil 1.00
Oyster shell 1.00
Mono calcium phosphate 1.50
DL-methionine 0.15
Sodium chloride 0.15
Mineral-vitamin premix* 0.50
Vitamin A 0.10
Vitamin E 0.10
Analysis results
ME (Kcal/kg) 2863.00
CP (%) 24.40
Calcium (%) 1.02
Available phosphorus (%) 0.59
Methionine (%) 0.57
Methionine +cyctine 0.93
Lysine (%) 1.54
*Supplemented for kg of the diets: Vit. A, 12000 IU; D3, 2000
IU; E, 20 mg; K3, 3 mg; B2, 7 mg; B3, 12 mg; B5, 3 mg; B12, 0.03
mg; biotin, 0.1 mg; choline chloride, 300 mg; Mn, 130 mg; Fe, 70
mg; Zn, 60 mg; Cu,12 mg; I,1 mg; Se, 0.2 mg, and adequate
antioxidant.
Table 2. Means growth performance values and mortality of quails fed additives at 42 day of age
Diets (treatments) Feed intake Body weight Feed conversion ratio Mortality
(g) (g/g) (%) (g/bird)
Basal diet (control group) 697±7b* 222±6bc 3.14±0.10a0.0
Basal diet + probiotic 701±4b220±7c3.19±0.11a0.0
Basal diet + prebiotic 706±3a226±4ab 3.12±0.08ab 0.0
Basal diet + synbiotic 706±3a229±4a3.08±0.07b0.0
*Figures bearing different superscripts within a row differs significantly (P<0.05).

872 BABAZADEH ET AL. [Indian Journal of Animal Sciences 81 (8)
92
control groups (P<0.05). Moreover, birds fed prebiotic
showed a significant increase (P<0.05) compared to birds
fed probiotic group (P<0.05). Results indicated that
consumption of synbiotic and then prebiotic were more
effective than other groups in BW, FI and FCR of Japanese
quail.
The present study demonstrated that the synbiotic and
prebiotic products displayed a greater growth-promoting
effect than the probiotic and control groups. The beneficial
effects of the synbiotic and prebiotic products on quail’s
performance parameters including FI, FCR, and BW are in
agreement with previous studies (Kabir et al. 2004,
Mountzouris et al. 2007 and Samli et al. 2007). The results
of present study about the effects of probiotic in BW, FCR
and FI are in agreement with the study of Maiolino et al.
(1992) which reported that probiotics did not have any
significant positive effect on broilers. Barrow (1992)
indicated that there was little evidence in the studies to
support the claims of positive effects made by probiotics and
the studies may have suffered from errors in methodology
and interpretation. Several studies showed that addition
prebiotic to poultry diets enhanced performance (Grimes et
al. 1997 and Rodriguez et al. 2005, Navidshad et al. 2010).
The male birds fed each one of additives showed a
significant decrease (P<0.05) in liver weight compared to
control group. Whereas, the female birds fed prebiotic
showed a significant decrease (P<0.05) in liver weight
compared to birds fed synbiotic and control groups (Table
3). These results are in agreement with the study of Azadegan
Mehr et al. (2007). Similarly, Mohan-Kumar and Christopher
(1988) reported a significant decrease in liver relative weight
due to lactobacillus and other beneficial microorganisms,
which are present in probiotics, can prevent pathogens from
colonizing the gastrointestinal tract via competitive
exclusion. With decrease in harmful microflora of intestine,
less toxic byproducts will be produced, so that the liver would
be under a less pressure for detoxifying these byproducts.
The heart weight significantly decreased (P<0.05) in the male
birds fed prebiotic-additive compared to synbiotic and control
groups. The female birds fed probiotic showed a significant
increase (P<0.05) in heart weight compared to other groups
(Table 3). Increasing heart weight in female birds fed
probiotic probably could be cause to heart hypertension or
appearing to ascites disease. Good management of present
experiment caused no disease and mortality (Table 3).
Effects of dietary treatments on GGT, ALP, CPK, AST,
ALT and LDH enzyme activities in male and female birds
are summarized in Table 4. Dietary additives did not have
any significant effect on activities of GGT and LDH enzymes.
GGT enzyme activity showed the lowest level of activity in
males fed probiotic compared to other groups and it showed
the lowest level of activity in females of control group
Table 3. The effect of dietary treatments on heart and liver weights (g) of quails at 42 days of age
Diets (treatments) Heart Liver
Male Female Male Female
(Percentage of live weight)
Basal diet (control group) 0.98±0.15a* 0.75±0.04b2.93±0.42a2.73±0.49a
Basal diet + probiotic 0.89±0.04ab 0.90±0.05a1.88±0.38b2.55±0.42ab
Basal diet + prebiotic 0.81±0.00b0.71±0.07b1.98±0.09b2.31±0.26b
Basal diet + synbiotic 0.92±0.07a0.71±0.06b1.70±0.14b2.81±0.46a
*Figures bearing different superscripts within a row differ significantly (P<0.05).
Table 4. Effects of feed additives on blood enzymes of Japanese quails at 42 days of age
Male female
Control Probiotic Prebiotic Synbiotic Enzymes Control probiotic Prebiotic Synbiotic
GGT 7.00±0.35a6.50±0.26a9.00±0.68a8.50±0.55a5.25±0.15a10.25±0.67a12.25±0.61a7.75±0.42a
ALP 1314.25 1513.50 1335.75 1816.75 2315.00 1713.25 1462.75 1600.75
±14.69b±19.76b±12.66b±25.36a±26.30a±33.44b±30.70b±18.28b
CPK 658.50 663.50 1145.50 844.50 687.50 728.00 1030.00 798.00
±27.26b±2.14b±7.76a±23.36b±25.29b±7.00ab ±16.78a±21.05ab
AST 234.00 259.50 292.25 267.50 274.50 284.50 278.75 283.00
±4.50b±3.28ab ±3.49a±2.52ab ±2.69a±2.82a±4.38a±2.84a
ALT 24.75 18.00 20.02 17.50 32.50 29.00 25.50 26.75
±0.34a±0.35b±0.38ab ±0.21b±0.87a±1.62a±80a±1.24a
LDH 632.75 637.75 841.50 653.25 637.25 606.75 673.25 616.75
±12.97a±20.43a±14.39a±28.88a±0.84a±11.98a±18.80a±18.02
*Figures bearing different superscripts within a row differs significantly (P<0.05).

August 2011] EFFECTS OF ADDITIVES INTAKE ON JAPANESE QUAILS 873
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compared to other groups. LDH enzyme showed lowest level
of activity in females fed probiotic compared to other groups
and it exhibited the lowest level of activity in males of control
group compared with other group. Effects of dietary additives
on ALP activity showed a highly significant difference
between male and female birds (P<0.01). The female birds
fed additives exhibited significantly low levels (P<0.05) of
ALP activity compared to the control group (P<0.05). The
biochemical analysis showed a significant increase (P<0.05)
in activity of ALP in male birds fed synbiotic compared to
the other groups. Similar results were reported by Hashem
and Mohamed (2009). Significant differences in serum levels
of ALP in male and female birds can be result of sexual
differences in Japanese quails. Higher ALP and CPK
activities in males than females could have been due to higher
osseous (ALP) and muscular (CPK) male development, just
as it occurs in most vertebrates (Coppo 2001). Effects of
dietary additives on ALT activity showed a significant
difference between male and female birds (P<0.05). The male
birds fed probiotic or synbiotic showed a significant decrease
in ALT activity (P<0.05) compared to control group. In
female birds, additive fed groups exhibited low levels of ALT
activity compared to control group. Male and female birds
fed additives showed increased AST activity compared to
control group. The male birds fed prebiotic showed a
significant increase (P<0.05) in AST activity compared to
control group. However, the female birds fed probiotic
showed an increase in serum levels of AST compared to
other groups but it was not a significant increase. The
male birds fed prebiotic showed significantly (P<0.05)
high levels of CPK activity compared to other groups.
Whereas, increase in CPK activity in other groups did not
have a significant difference compared to control group. The
female birds fed prebiotic showed a significant difference
(P<0.05) in serum levels of CPK activity compared to control
group.
Present study indicated that feed additives caused to higher
levels of AST and CPK activities and prebiotic caused the
high levels of GGT and LDH activities in serum of male and
female birds compared to control group. In an investigation
Imaeda (2000) indicated that increase serum level of enzyme
utilized as indicators for clinical diagnosis of cardiac failure
is in association with sudden death syndrome (SDS). Studies
demonstrated that CPK, LDH, and AST activities
significantly increased in the serum of broilers chickens that
died by SDS (Imaeda 1999, Durdi and Aliakbarpour 2005).
The feed additives and especially prebiotic can be caused a
circulatory enzymes elevation. Since these enzymes are not
heart-specific, for definitive diagnosis we should account
the other laboratory data and clinical observations carefully.
Ozyurt et al. (2006) reported that AST, ALT, GGT and LDH
usually appear in serum when there is damage on the liver
and muscle tissues caused by excessive stress. Cyber (1999)
and Panda et al. (2000) reported that probiotic has been
promoted to reduce stress. This study demonstrated that
probiotic caused low AST, GGT and LDH activities in serum
of male birds compared to synbiotic and prebiotic, and it
caused significant decrease (P<0.05) in serum level of ALT
in male birds compared to control group. It can be concluded
that probiotic by decreasing effects of stress can cause a lower
enzyme activity and it can be a protective agent for liver and
muscles against damage factors in male quails compared to
groups fed other additives. Furthermore, consumption of all
feed additives decreased ALT activity in male and female
birds compared to control group and they can help in health
of liver and muscles as a protector agent.
The present study demonstrated that the synbiotic and
prebiotic displayed a greater growth-promoting effect than
the probiotic, and the control group. Synbiotic seemed more
effective in performance of Japanese quail than other
additives. Consumption of prebiotic in males and females
reduced in heart weight that is important factor for animal
health. Probiotic intake can cause a lower enzyme activity
and decreasing effects of stress. Thereby it can be a protective
agent for liver and muscles against damage factors in male
quails compared to other feed additives. In addition to
diagnosing enzymatic activity, and of heart weight enhancing
in female Japanese quail fed probiotic will be stimulating
for more studies.
ACKNOWLEDGMENTS
The authors would like to thank Damavand Quail Co.
(www.dquail.ir) Tehran-Iran for equipments of the
Research Room of Japanese Quail in Tabriz-Iran and
The Etouk Farda Co. (www.etouk.com) Tehran-Iran for
providing Biomin® IMBO and Javaneh Khorasan Co.
(www.javanehkhorasan.com) Mashhad-Iran for supplying
Fermacto® for facilitating to complete present research
project.
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