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CHAPTER-1
INTRODUCTION
The sub-therapeutic use of antibiotics in livestock and poultry production is under
severe scientific and public scrutiny, as antibiotic growth promoters (AGP) are linked
with the development of pathogenic bacteria which are antibiotic-resistant. These
pathogenic bacteria create health problems (Smith et al., 2003). As a result, the European
Union banned on sub-therapeutic usage of AGP in animal production in 2006 (Burch,
2006). Due to impending ban of AGP in livestock and poultry feed, it was compulsory
for poultry industry to develop alternatives of AGP. The prebiotics and probiotics seem to
be alternate candidates for AGP (Cavazzoni et al., 1998).
Prebiotics are the feed ingredients that are not digested by host digestive enzymes
instead are fermented by beneficial bacteria and, therefore, are beneficial for host (Gibson
and Roberfroid, 1995). Oligosaccharides fall under this category and are believed to
affect the gut health of host (Ferket, 2004). Mannan-oligosaccharides (MOS), extracted
from yeast cell wall, are not hydrolyzed by the host enzymes and are fermented by
intestinal microbiota (Flickinger and Fahey, 2002). Mannan-oligosaccharides provide
competitive binding sites for pathogens with mannose-specific type-1 fimbriae such as
salmonella and E. coli and decrease their attachment with intestinal wall and are
ultimately excreted from the gut (Newman, 1994; Ferket et al., 2002). It has been
demonstrated that MOS supplementation constantly increases the cecal populations of
1
Lactobacillus spp. and Bifidobacterium spp. (Yang et al., 2009; Oyofo et al., 1989;
Spring et al., 2000; Baurhoo et al., 2007).
Prebiotics have been shown to have a positive effect on growth performance in
poultry. Prebiotics improved body weight and feed conversion efficiency of turkeys
(Sims et al., 2004; Fritts and Waldrop, 2003). Hooge et al. (2003) investigated that
dietary MOS supplementation has significant improvement in body weight, feed
conversion ratio in broilers without any effect on mortality.
Prebiotics, especially, oligofructose, gluco-oligosaccharide, and galacto-
oligosaccharide have been found to stimulate absorption of several minerals, particularly
magnesium, calcium, and iron in rats (Scholz-Ahrens et al., 2001). Van den Heuvel et al.
(1998) investigated the effect of these prebiotics on calcium and iron absorption at a
much lower dose in healthy, adult human. Neither inulin nor the fructo- or galacto-
oligosaccharides increased calcium or iron absorption. Coudray et al. (1997) found that
inulin increased calcium absorption in man, while had no effect on the other minerals.
Ghosh et al. (2008) also reported that MOS had no significant effect on plasma minerals
of Japanese quail except Ca level that was higher in MOS-supplemented birds compared
to control birds.
Very little data is available regarding growth-promoting effects of MOS in
Japanese quail and on the mineral absorption. Keeping in view the existing knowledge it
is hypothesized that MOS supplementation can enhance mineral absorption and improve
growth performance of Japanese quail.
OBJECTIVE
2
The present study was conducted to investigate effects of Mannan-
oligosaccharides (MOS) supplementation on production performance, cecal microbial
population and mineral absorption in Japanese quail.
CHAPTER-2
LITERATURE REVIEW
PREBIOTICS
Prebiotics are non-digestible food ingredients that beneficially affect the host by
selectively stimulating the growth and /or activity of one or a limited number of bacteria
(Gibson et al., 2004). Prebiotics modify the composition of the intestinal microbiota,
especially health promoting bacteria, lactobacilli and bifidobacteria which improve the
host’s health. In order for a food ingredient to be considered a prebiotic, it must have
following properties.
It must be neither hydrolyzed by host enzymes nor absorbed in the upper part of
gastrointestinal tract.
It must be selectively fermented by one or a limited number of beneficial bacteria.
It must alter the intestinal microbiota and their activities in the host.
It must preferably induce effects that are beneficial to the host health.
(Gibson and Roberfroid, 1995 and Patterson and Burkholder, 2003)
The fermentable substances that can acts as prebiotics are non-starch
polysaccharides, dietary resistant starch, and non digestible oligosaccharides (Piva et
3
al., 1996; Jacobasch et al., 1999). The most dominant candidates for acting as
prebiotics are non-digestible oligosaccharides (Bauer et al., 2006).
MANNAN OLIGOSACCHARIDES
Carbohydrates are important structural components of the majority of cell-surface
and secreted proteins of animal cells (Osborn and Khan 2000). Oligosaccharides are
formed when 2-10 monosaccharide molecules are joined together to form a larger
molecule. More than 10 monosaccharide molecules joined together to make a
polysaccharide. Mannose is a monosaccharide that forms the building block of Mannan-
oligosaccharides (MOS). Mannose-based oligosaccharides occur naturally in cell walls of
the yeast Saccharomyces cerevisiae and obtained by centrifugation of lysed yeast culture
(Spring et al., 2000). The commercially available product Bio-Mos® (Alltech, Inc.,
Nicholasville, KY) is a source of MOS from Saccharomyces cerevisiae cell walls. This
product was introduced in 1993 as a feed additive for broiler chickens (Hooge, 2003).The
small intestine does not contain the digestive enzymes required to break down mannan-
oligosaccharide bonds, therefore they arrive at the large intestine intact after ingestion
and passage through the small intestine (Strickling et al., 2000). A proposed mechanism
of prebiotic action is given in Fig.1.
4
Fig. 1: A proposed mechanism of prebiotic action to improve health (Crittenden, 1999)
5
Scholz-Ahrens et al. (2001) studied effects of prebiotics on mineral metabolism.
Non digestible oligosaccharides (NDO) have been found to stimulate absorption of
several minerals and to improve mineralization of bone. The scientific evidence for the
functional effects of NDO is based on animal experiments in which NDO increased the
availability of calcium, magnesium, zinc, and iron. This stimulatory effect of some NDO
is assumed to be mainly due to their prebiotic character. It stimulates the growth and
activity of bacteria with beneficial effects on health of the host. These findings were also
confirmed in human studies. The effects seem to be specific for the type of carbohydrate
and are likely related to the rate of fermentation by the intestinal flora and appear to
depend on the ingested dose.
Fairchild et al. (2001) studied the effects of hen age, Escherichia coli, and
dietary Bio-Mos and Flavomycin on poult performance. Day-of-hatch BUTA (BIG-6)
male poults were gavaged orally (1 mL) with approximately 10(8) cfu/mL E. coli
composed of four serotypes or sterile carrier broth. A mixture of the same E. coli cultures
was added to the poult’s water troughs to attain a concentration of approximately 10(6)
cfu/mL on a weekly basis to ensure a continuous bacterial challenge. Within each E. coli
split plot treatment group, poults from hens of different ages were fed diets containing
Bio-Mos, Flavomycin, Bio-Mos plus Flavomycin, or a control diet, in a randomized
complete block design. This experiment yielded eight treatments per challenge group.
During E. coli challenge, dietary Bio-Mos and Flavomycin improved poult BW and BW
gains. When poults were not challenged with E. coli, poults from old hens had improved
BW and cumulative BW gains over poults from young hens. Cumulative 3-wk BW gains
for unchallenged poults from young hens were improved by Bio-Mos and Flavomycin
6
alone and in combination when compared to the control diet. It may be concluded that
dietary Bio-Mos and Flavomycin can improve the overall performance of poults,
especially when they are faced with an E. coli challenge.
Fernandez et al. (2002) conducted different studies to investigate the effects of
mash diet, or mash supplemented with either MOS or palm kernel meal (PKM) and
caecal contents of hens (HCC) fed with mash on the major microflora groups of chicks,
and their inhibitory effect on Salmonella colonization and the effect over time of diets
supplemented with MOS or PKM on S. Enteritidis colonization and the microflora of
chicks. In hens, supplemented diets increased Bifidobacterium spp., while decreasing
members of Enterobacteriaceae and Enterococcus spp., compared with the mash diet.
Chicks dosed with the HCC showed, on average, increased numbers of anaerobes, while
the numbers of aerobes decreased including coliforms and S. Enteritidis compared with
controls without HCC. In chicks fed the MOS-supplemented or PKM-supplemented
diets, S. Enteritidis colonization decreased over time, compared with mash alone. Four-
week-old PKM birds showed an increase in Bifidobacterium spp. and Lactobacillus spp.,
with a decrease in S. Enteritidis compared with week 2. Generally, the HCC and diets
supplemented with MOS or PKM affected the bird’s intestinal microflora by increasing
the Bifidobacterium spp. and Lactobacillus spp., while decreasing the Enterobacteriaceae
groups. They also reduced susceptibility in young chickens to colonization by S.
Enteritidis.
Spais et al. (2003) studied effect of the mannan-oligosaccharide on broiler
performance. A total of 53,040 one day-old Cobb chicks, randomly divided into two
groups were used in a feeding trial that lasted 40 days. One of the groups was fed on a
7
basal commercial starter diet, while the other was given up to day 10 of age the same diet
supplemented with the mannan-oligosaccharide Bio-MOS at the level of 1.5 g/kg of feed.
From day 11 of age and thereafter, Bio-Mos administration was discontinued and both
groups were given the same basal commercial grower and finisher diets. Results showed
that chickens in the Bio-Mos fed group exhibited a significant (P<0.05) improvement in
body weight compared to control at day 10 and day 40 of age. Feed intake per bird and
feed conversion ratios demonstrated a significant (P<0.05) improvement for the Bio-Mos
group. Mortality rate was lower in the Bio-Mos group compared to control, however, the
difference was not statistically (P>0.05) significant.
Parlat et al. (2003) conducted an experiment to evaluate the effects of mannan-
oligosaccharides (MOS) or Virginiamycin (VM) on the growth performance of Japanese
quails. The quails were assigned to 4 dietary treatments: Control, MOS, VM or
MOS+VM. Individual body weight and feed consumption were recorded weekly.
Mortality was recorded when occurred. All treatments significantly (P<0.05) increased
body weight for 5 wk, and improved feed conversion ratio for 0-3 and 3-5 and 0-5 wk.
There were no treatment effects for feed consumption during trial. Dietary supplemental
MOS, VM or MOS+VM resulted in improved growth performance of Japanese quails.
These results indicate that MOS may be utilized as an alternative to antibiotic growth
promoter to improve the quail performance.
Hooge et al. (2003) conducted a study to compare the efficacy of commercial
mannan oligosaccharide (MOS) as an alternative growth promoter to Bacitracin
Methylene Disalicylate (BMD) followed by virginiamycin (VM). Feed phases were 0 to
21, 21 to 42, and 42 to 49 d. In experiment 1, treatment effects were non significant at 21
8
d. At 49 d, BMD or MOS significantly (P < 0.05) improved body weight and feed
conversion ratio and increased feed expense per bird and net income per bird, without
affecting mortality, compared with control group.
In experiment 2, there were 6 dietary treatments: The BMD + MOS, VM + MOS
shuttle program gave best body weight, feed conversion, and mortality at 21 and 49 d of
age resulting in the lowest feed expense and highest net return per bird. It was concluded
that MOS supported live performance equivalent to BMD followed by VM and had an
additive effect when combined with the antibiotics.
Hooge (2004) studied the global broiler chicken pen trial reports (1993-2003) and
analyze statistically to determine effects of mannan-oligosaccharide (Bio-Mos)
supplemented diets versus negative control (nCON) or antibiotic supplemented positive
control (pCON) diets. Results were averaged "by treatments" and "by trials" using Paired
T-test to compare nCON and pCON means with corresponding MOS means. Slightly
different answers but similar patterns emerged by these methods.
Considering results averaged by trials, MOS diets improved the BW and lowered
mortality compared to nCON diets. Relative improvements using MOS feeds compared
to the pCON diets were non significant. The MOS diets significantly (P = 0.008) lowered
mortality relative to pCON diets, indicating a strong beneficial effect. The MOS diets
improved BW and FCR comparable to those of pCON diets but significantly lowered
MORT compared to antibiotic diets.
Tarasewicz et al. (2004) studied influence of oligosaccharides isolated from pea
seeds on functional quality of quail. The birds were divided into three feeding groups
(two replications) of 48 female and 16 male birds each. Quail of the first group were fed a
9
standard feed, those of group 2 and 3 received feed enriched with oligosaccharides at a
dose of 0.4 g and 3 g, respectively. The oligosaccharide-enriched feed reduced the time
of maturation, increased egg laying capacity and egg weight, and also decreased the
consumption of feed per egg. No clear influence of the oligosaccharide supplementation
was found as far as the blood cholesterol and triglyceride content was concerned and
gammaglobulin in the eggs. The quail of the groups receiving oligosaccharides had lower
bifidobacteria counts in their digestive tracts.
Sims et al. (2004) studied effects of dietary mannan oligosaccharide, Bacitracin
Methylene Disalicylate (BMD), or both on the live performance and intestinal
microbiology of turkeys. Four dietary treatments were used: one negative control (CON)
and other three diets formulated with different levels of MOS and BMD. The BMD and
MOS turkeys were heavier than CON birds, and those fed the combination were
significantly heavier than all other treatments. At wk 18, BMD + MOS feed conversion
ratio was significantly lower than CON and with BMD and MOS being intermediate.
Mortality was not affected by treatment. The BMD and MOS each reduced large
intestinal concentrations of Clostridium perfringens at wk 6 but not at wk 18. The BMD
or MOS each improved turkey performance, and when used together, exhibited further
beneficial effects.
Oguz and Parlat (2004) studied effects of dietary mannan oligosaccharide on
performance of Japanese quail affected by aflatoxicosis. The potential of the mannan
oligosaccharide (MOS) to ameliorate the effects of aflatoxicosis was examined in
growing Japanese quail. The product was incorporated in the diet at 1 g/kg and was
evaluated for its ability to reduce the deleterious effects of 2 mg total aflatoxin /kg diet on
10
Japanese quail chicks from 10 to 45 days of age. Forty 10-d old Japanese quail chicks
were assigned in a 2x2 factorial arrangement of treatments to four groups (Control, AF,
MOS, AF plus MOS), each consisting of 10 quails. The addition of AF alone
significantly decreased feed consumption and body weight gain from the first week
onwards. A significant adverse effect of AF on the feed conversion ratio was also
observed from week 4 onwards. The addition of MOS to the AF-containing diet
significantly reduced these adverse effects of AF on feed consumption, body weight gain
and feed conversion ratio. The cumulative body weight gain was 22.0% lower in the
quails consuming a diet containing AF without MOS as compared to the control group.
However, it was only 2.3% lower that the control in the birds fed the diet containing the
AF plus MOS.
Flemming et al. (2004) carried out a study with 2,400 broilers to compare the
effect of the use of mannan-oligosaccharides, Saccharomyces cerevisiae cell wall or
growth promoter (Olaquindox) in the diet on broiler. Diets were based on corn and
soybean meal. A completely randomized experimental design was used, and the obtained
data was evaluated by analysis of variance and test of Tukey at a level of 5%. Feed
intake, daily weight gain, feed conversion ratio, and mortality were measured. It was
concluded that the effect of the inclusion of mannan-oligosaccharides in the diet on the
studied parameters was significantly higher as compared to the inclusion of cell wall or to
the control diet, but the effect was not different as compared to the inclusion of growth
promoter.
Parks et al. (2005) studied effects of virginiamycin and a mannan-
oligosaccharide-virginiamycin shuttle program on growth performance, body weight
11
uniformity, and carcass yield characteristics of large white female turkeys. Diets
containing no growth promoter, VM, or a shuttle program of MOS and VM were fed to
Hybrid female turkeys.
Body weights and feed consumption were recorded at 3-wk intervals, and
mortality and culled birds were recorded daily. At the conclusion of the trial, 2 birds per
pen were randomly chosen for carcass yield analysis. Feeding VM alone significantly
increased body weight compared with control fed birds during all periods. The MOS-VM
shuttle program resulted in early growth depression for birds less than 3 wk of age,
possibly influenced by an unplanned cold stress, but better growth than the non
medicated control birds after 6 wk of age. Birds fed VM had superior (P < 0.05) feed
conversion ratio from 0 to 3 wk, which persisted until 14 wk (P < 0.10). There were no
treatments effects on overall feed consumption, uniformity, mortality, or cull rate.
Processing yields or weight of various parts were also unaffected by treatment.
Blake et al. (2006) conducted a series of four consecutive studies on built-up litter
to compare efficacy of a commercial mannan-oligosaccharide (Bio-Mos) and BMD when
broilers were fed wheat based diet. In each trial a total of 1500 broiler chicks were
obtained. Built-up litter was used throughout with one flock reared on the litter prior to
trial initiation and experimental groups were maintained on litter from the same
treatments with no top dressing between flocks. Broilers were subjected to three
treatments, control, Bacitracin Methylene Disalycilate (BMD) or mannan oligosaccharide
(MOS). Birds were fed starter, grower and finisher diets. Diets were corn-wheat soybean
meal based to include 30% wheat and 600 units/ton xylanase. Coban was used in starter
and grower diets. Diets and water were ad libitum and light was 23D:1L. Birds and feed
12
were weighed at 14, 28 d and at a target weight of 2.2 kg. Results from combined data
analysis indicate highly significant (P < 0.0009) improvements in BW with MOS and
BMD over CON at 14 d. These differences diminished by 28 and 37 d, but MOS and
BMD showed numerically greater improvements in body weight. Feed consumption at 14
d was greatest for BMD intermediate for MOS and lowest for CON, after which no
differences in FCR were noted. Results indicate that the addition of Bio-Mos to the diet
had an influence in promoting bodyweight increases over the control diet early in the
growing period, typically from the 0-14 d period. The combination of continued use and
long-term effects indicate that cumulative improvements in performance may be
attributed to the use of specific feed additives such as Bio-Mos.
Solis et al. (2007) studied effect of Alphamune, mannan-oligosaccharide in turkey
poults. Two trials were conducted to evaluate the effects of Alphamune on gut maturation
of 7- and 21-d-old turkey poults. Poults were fed a standard control unmedicated turkey
starter diet or the same diet supplemented with Alphamune. Poults were weighed on d 7
and 21, On d 7, BW was higher for the poults given the Alphamune treatments compared
with control poults; however, no differences were observed on d 21.
Baurhoo et al. (2007) conducted a study to evaluate lignin and mannan
oligosaccharides as potential alternatives to antibiotic growth promoters in broilers.
Dietary treatments included an antibiotic-free diet (CTL–), a positive control (CTL+), and
an antibiotic-free diet containing Bio-Mos or Alcell lignin. Body weight and feed
conversion were recorded weekly. Cecal contents were assayed for Escherichia coli,
Salmonella, lactobacilli, and bifidobacteria, and the litter was analyzed for E. coli and
Salmonella. Birds fed the CTL– diet were heavier (P < 0.05) than those fed the other
13
dietary treatments, but feed conversion was not affected by dietary treatments. Birds fed
MOS had greater lactobacilli numbers than those fed the CTL+ diet and also increased the
populations of bifidobacteria in the ceca. Litter E. coli load was lower in birds fed MOS
than in birds fed the CTL+ diet. Broiler performance was similar in birds fed antibiotics
or antibiotic-free diets containing either MOS or lignin.
Ghosh et al. (2007) conducted an experiment to determine the effect of dietary
supplementation of organic acid and mannon-oligosaccharide on the performance and gut
health of Japanese quail. Day old chicks of Japanese quail (n=280) were randomly
assigned into seven dietary treatments replicated four times with ten chicks per replicate.
Control (Co) birds were given a standard basal diet; and diets for T1-T6 birds will be
formulated with different levels of MOS (prebiotic) and organic acid salts (OAS).
Statistical analysis reveals that OAS supplementation increased live weight, live weight
gain compared to control (C).Cumulative feed intake was not significantly affected due to
dietary treatments. Superior results in term of feed conversion ratio (FCR) and
performance index (PI) were found in MOS supplemented groups compared to others.
Organic acid salts with MOS improved gut health by reducing bacterial load compared to
control and other groups.
Yang et al. (2007) conducted a trial to study influence of MOS on growth
performance and bacteriological, morphological and functional aspects of small intestine
in broiler chickens at different ages. Three dietary treatments were used: a negative
control without MOS, a positive control (Zn Bacitracin), and 2 g of MOS/kg of diet. The
MOS supplementation has improved BW gain compared with the negative control in
early life. Total anaerobic bacteria, lactic acid bacteria, and Clostridium perfringens were
14
not affected by the supplementation of MOS. Coliform bacteria were increased in young
birds treated with MOS. In the current study conducted under hygienic experimental
conditions, the addition of MOS did not show a clear positive effect on performance of
broilers.
Yang et al. (2008) studied effects of mannan-oligosaccharide (MOS) on the
growth performance, nutrient digestibility and gut development of broilers given a corn
or a wheat-based diet over a 21-day experimental period. Dietary MOS improved the
growth performance of birds given the wheat-based diet compared to that of birds given
the corn-based diet during 7-21 days of age. The addition of MOS modulated the
development of gut microflora. From day 7 to day 21, the numbers of mucosa-associated
coliforms along the small intestine were decreased; whereas the numbers of mucosa-
associated lactobacilli were increased by MOS. Dietary MOS also reduced the counts of
coliforms and Clostridium perfringens in the ceca of birds by 21 days of age. All these
changes were dependent on the type of cereal and the age of the birds.
Yang et al. (2008) studied effects of mannan-oligosaccharide on the growth
performance and digestive system, particularly gut microflora using 1-d-old birds in an
Escherichia coli challenge model. The experiment lasted for 3 weeks and zinc bacitracin
(ZnB) was used as a positive control. Statistical analysis showed that dietary MOS had
positive effects on body weight gain (BWG) and feed conversion efficiency (FCE) of the
challenged birds compared to the negative control. Similar results were obtained for ZnB
treatment. The addition of MOS reduced the number of mucosa-associated coliforms in
the jejunum of the challenged birds on d 7. The number of Clostridium perfringens in the
gut lumen was reduced by only ZnB. In conclusion, the effects of MOS on the
15
composition and activities of gut microflora and mucosal morphology of birds were
related to E. coli challenge as well as the age of birds, which may be involved in the
observed different growth-improving effects of the tested dietary additives.
Ghosh et al. (2008) conducted an experiment to determine the influence of
organic acid salts (OAS) and MOS on carcass traits and plasma minerals of Japanese
quail. Day old chicks of Japanese quail (n=280) were randomly assigned into seven
dietary treatments replicated four times with ten chicks per replicate. Control birds were
given a standard basal diet; and diets for T1-T6 birds were formulated with different
levels of MOS and organic acid salts. Statistical analysis reveals that OAS and MOS had
non-significant effect on carcass traits and plasma minerals except calcium level which is
varied significantly among the experimental groups due to dietary treatments.
Benites et al. (2008) conducted a trial on broiler chickens to evaluate the effects
of dietary mannan-oligosaccharide (MOS) from either of 2 commercial products, Bio-
Mos or SAF-Mannan, each at 2 levels of inclusion on live performance. Diets were fed in
3 phases, and treatments included a control, 2 Bio-Mos treatments, and 2 SAF-Mannan
treatments, Birds fed Bio-Mos diets had significantly greater BW at 42 d than birds fed
control or SAF-Mannan-supplemented diets, whereas results for Feed consumption was
lower from 0 to 21 d in the SAF-Mannan treatments compared with other treatments. No
significant differences were found for feed conversion or mortality for any of the
treatments. Overall, Bio-Mos had a greater effect on bird BW compared with the other
variables measured.
Mohamed et al. (2008) performed an experiment in which natural growth
promoter (MOS) was compared with an antibiotic growth promoter (enramycin) on
16
performance and carcass characteristics of broiler chicks. Four treatment groups were
made which are, a diet of no supplement served as a control, basal diet with MOS (1g/kg)
and basal diet +Enramycin (0.35g/kg) while another diet was supplemented with both
MOS and Enramycin. The dietary treatments were fed to four replicates of 15 chicks
each. The results indicated that addition of MOS, enramycin or the combination of both
did slightly improve body weight gain compared to the control diet. Feed conversion ratio
were significantly improved by the addition of MOS, enramycin or the combination of
both. No significant effects on liver, heart and gizzard weight were detected. It is
concluded, that MOS might be used as an alternative to growth-promoting enramycin in
broiler diets.
Sahin et al. (2008) carried out an experiment to determine the effect of dietary
supplementation of combiotics (probiotics + prebiotics +makrotone) on body wt gain,
feed consumption and feed conversion ratio. A total of 264 daily Japanese quail chicks
(coturnix coturnix japonica) were used in the experiment. They were divided in 1 control
and 3 treatment groups each containing 66 chicks. The experimental period lasted for 35
days. Control group was fed with supplemental basal diet. 0.5, 1.0, and 1.5g/kg combiotic
was added to diet of treatment groups 1, 2 and 3 respectively. At the end of experiment,
the effects of combiotic supplementation to diet on the BWG, FC and carcass yield of
quail were not statistically significant among the groups (p>0.05).
Bozkurt et al. (2008) investigated the effect of dietary supplementation with an
antibiotic growth promoter (AGP) and two prebiotics; mannan oligosaccharide (MOS)
and dextrin oligosaccharide (DOS), respectively, on growth performance of broilers. One
thousand and two hundred day-old broiler chicks (Ross 308) were assigned to the four
17
treatment groups. The four treatments were as Basal diet, Basal diet + antibiotic, Basal
diet + mannan oligosaccharide (1 g/kg diet), Basal diet + dextran oligosaccharide (1 g/kg
diet). Body weight of birds given MOS supplemented diet was significantly higher than
those birds fed with AGP and DOS added diets. Feed consumption, feed conversion ratio
of birds were not affected by dietary treatments. The results obtained in the present
experiment showed that birds fed with AGP, MOS and DOS supplemented diets
exhibited higher body weight gain.
Bozkurt et al. (2009) conducted an experiment in which the effects of some
alternative feed additives for antibiotic growth promoters on performance and some
slaughter characteristics were examined in broilers. A total of 2160 one-day-old male
broiler chicks were randomly allocated to six groups with six replicate pens per
treatment. The treatments were the basal diet (Control), and the basal diet supplemented
with an antibiotic growth promoter (AGP); a prebiotic, mannan-oligosaccharide (Bio-
Mos, MOS); an essential oil of oregano (Herb-Mos Oregano, HMO); a plant extract of
hop (Herb-Mos Hops, HMH) or a mixture of Herb-Mos Oregano and Herb-Mos Hops
(HMOH). There were significant effects of dietary treatments on body weight, feed
consumption and feed conversion ratio. The addition of all experimental additives to the
diet resulted in significantly higher body weights as compared to the control treatment.
Feed intakes and feed conversion ratios were significantly better at 0 - 21 d, but not
during the 0 - 42 d period. These results showed that AGP, MOS and herbal feed additive
supplementation to a diet provided significant advantages on broiler growth performance
through a 42-d growth period. However, the combined supplementation of HMO and
18
HMH did not exert either synergistic or additive benefits on the live performance of the
broilers.
Baurhoo et al. (2009) conducted an experiment in which the effects of 2 levels of
mannan oligosaccharide (MOS) in feed were compared with antibiotic growth promoters
on growth performance, cecal and litter microbial populations, and carcass parameters in
broilers raised in a sanitary environment. Dietary treatments included Basal diet (control),
basal diet +VIRG (virginiamycin), basal diet +BACT (bacitracin), LMOS (basal diet +
0.2% MOS), and HMOS (basal diet + 0.5% MOS). Body weight and feed intake were
recorded weekly. At the same bird ages, cecal contents were assayed for lactobacilli,
bifidobacteria, Salmonella, Campylobacter, and Escherichia coli, whereas litter was
analyzed for Salmonella, Campylobacter, and E. coli. Body weight and feed conversion
ratio did not differ among treatments. Bifidobacteria concentrations were higher (P <
0.05) in LMOS- and HMOS-fed birds at all time points. Birds and litter from all
treatments were free of Salmonella. In comparison to birds fed control, BACT, LMOS,
and HMOS significantly reduced (P < 0.05) cecal E. coli concentrations, Litter bacterial
counts were not altered by dietary treatments. In conclusion, under conditions of this
study, MOS conferred intestinal health benefits to chickens by improving its
morphological development and microbial ecology. But, there were no additional benefits
of the higher MOS dosage.
Markovic et al. (2009) performed an experiment to study effects of different
growth promoters on broiler performance and gut morphology. A total of 240 Hybro
broilers were divided into 4 groups. These groups were fed a complete soybean based
diet with and without addition of antibiotic growth promoters (AGP, Flavomycin), direct
19
feed microbial (DFM, All-Lac) and mannan-oligosaccharide (Bio-MOS). At day 42 of
trial, all broilers were conventionally sacrificed in a slaughter plant and slaughter
performances were measured. At the end of the trial, body weight (BW) and body weight
gain (BWG) of broilers fed the diet containing Bio-MOS, AGP and DFM were
significantly higher and lower FCR than in birds of the control group. In conclusion, Bio-
MOS® and DFM exhibited nutritional, pharmacological and economic advantages over
antibiotic growth promoters.
Eleftherios et al. (2010) conducted a trial to see the effect of the dietary
supplementation of Mannan-oligosaccharides (MOS) and the acidifier Calcium
Propionate (CP) on the performance and carcass quality of the Japanese quail. They took
300, one-day old Japanese quail and divided into four groups with three subgroups, each
were fed a basal diet that served as control, or a basal diet with 6 g/kg CP, or 1 g/kg MOS
or 1 g/kg MOS plus 6 g/kg CP. The body weight, feed consumption, feed conversion
ratio and mortality of the birds were calculated at weekly intervals. The results of the
experimentation showed that the addition of MOS in the feed of growing quail
significantly increased the body weight on second week and the feed consumption on
second and fourth weeks, while it decreased the liver to live weight percentage. No
adverse effects from the dietary addition of MOS or CP or both were observed on the
performance or the carcass quality of the growing quail.
20
CHAPTER-3
MATERIAL AND METHOD
The study was conducted to investigate effects of dietary Mannan-
oligosaccharides (MOS), (Bio-Mos® by Alltech, Inc. USA) on the growth performance
of Japanese quails. A trial was conducted at the Avian Research and Training Centre,
University of Veterinary and Animal Sciences, Lahore, Pakistan.
Experimental Birds and Management
A total of 1,280 day-old Japanese quails (Coturnix coturnix japonica) chicks,
procured from the hatchery of Avian Research and Training Centre and randomly divided
into 4 groups (A, B, C and D). Each group was consisting of 320 birds and further
replicated into eight groups (n = 40) in a completely randomized design. The birds were
housed in wire-bottomed battery equipped with bulbs for light during the 35 days of
experimental period. Jute bags were used as bedding material during day 1 to day 12. The
trial was conducted in a closed shed with proper ventilation. The initial temperature of
the house was maintained at 37oC during first week of the experiment and was gradually
reduced according to normal management practice (1-2oC/week) to 32 °C in the fifth
week. Chicks were maintained on a 24hr constant light schedule during the 35 day
experimental period. The birds were weighed weekly until the end of the experiment.
Feeding & Diets
21
Birds were fed a corn-based basal diet, or the same basal diet supplemented with
MOS either at 1% (Group B), or 0.5% (Group C) or 0.1% (Group D) levels. Diet in group
A was in accordance with the nutritional requirements of Japanese quail as specified in
NRC (1994) and was considered as control (Table 3.1, 3.2). No coccidiostats or
antibiotics were added in the feed. Water and feed were provided ad libitum throughout
the experiment.
Table 3.1: Ingredient Composition of Experimental Diet
Ingredients Ingredients % (Diet A control )
Maize 30.0
Rice polish 8.00
Canola meal 10.0
Soybean meal 25.0
Corn gluten 60 % 5.00
Rice tips 14.0
Lime stone 1.00
D-L Methionine 0.10
L-Lysine 0.20
Threonine 0.15
Soy oil 1.85
DCP 1.50
Vitamin Premix 0.20
Molasses 3.00
22
Table 3.2: Nutrient Composition of Experimental Diet
Nutrients Nutrients % (Diet A control )
ME Kcal/kg 2900
CP 24.00%
Ca 0.80%
Available P 0.30%
Phytate P 0.34 %
Total P 0.65%
Crude fiber 4.38%
Linoleic acid 1.00%
Methionine 0.50%
Lysine 1.30%
ZOOTECHNICAL PARAMETERS
Feed Consumption
The weighed quantity of feed was offered to each experimental group. Residual
feed and left over feed was recorded to determine weekly feed consumption of each
group. At the end of experiment the overall feed consumption was calculated by adding
weekly feed consumption.
Body Weight Gain
The day-old chicks were weighed on day-1 and later subsequently on weekly
basis to calculate weekly body weight gain. The weekly weight gain was calculated by
23
subtracting the body weight of previous week from the body weight of next week. At the
end of experiment the total body weight gain was also calculated.
Feed Conversion Ratio
Recorded feed consumption and average body weight gain estimated on weekly
basis were utilized to calculate feed conversion ratio (FCR) according to the following
formula.
Feed consumed FCR = Body weight gain
SLAUGHTERING AND SAMPLING
On 35th day, 16 birds (2 per replicate per group) were randomly selected, weighed
and slaughtered for sampling. Blood samples were collected in heparinized vacutanors
and centrifuged at 3000 x g for 10 minutes to collect serum. Serum samples thus
collected were stored in aseptic plastic tubes at -20OC. The small intestine was
eviscerated to measure its length. The weights of gizzard and ceca, with and without
digesta were determined. After the removal of digesta from gizzard and cecum, the
tissues were washed thoroughly with ice-cold water, blotted and then weighed again. The
weights of the heart, and the liver were determined immediately after slaughtering. The
cecal digesta was collected in glass tubes in an ice beaker for enumeration of bacteria.
PARAMETERS STUDIED
WEIGHT AND LENGTH OF VISCERAL ORGANS
Weights of liver heart, ceca, gizzard and small intestine were recorded
24
immediately after slaughtering. Weights of gizzard, ceca and small intestine were
recorded with or without digesta. Similarly, length of small intestine was recorded with
or without digesta. Weights obtained thus were utilized to calculate relative weight of
organ by dividing it with the live weight of that bird as
Weight of Organ Relative weight = × 100 Live weight of bird
MINERAL ESTIMATION
Serum levels of calcium, magnesium, copper and iron were estimated by atomic
absorption spectrophotometer (Perkin Elmer, A Analyst-100) following the method
described by Sandal (1950) and modified by Arenza et al. (1997) while Phosphorous
analyzed by using spectrophotometer (AOAC, 2001).
MICROBIAL POPULATIONS OF CECAL DIGESTA
Cecal digesta thus collected was utilized for enumeration of bacteria. A sample of
0.5g of cecal material was added to saline solution and mixed. 1 ml of this solution was
transferred to next tube and so on to make 1:10 dilution as described by Miles and Misra
(1938) with some modification. Colonies of microorganisms were counted and multiplied
with the dilution factor to get number of live bacteria present in 0.5g digesta and results
were presented as log 10.
Clostridium perfringens culture
Medium preparation
Reinforced clostridial medium (RCM; Cat: 1007; Laboratories CONDA, Madrid,
Spain) was used to prepare culture medium. Reinforced clostridial medium, 38g and 2g
of agar were dissolved in 1 liter of de-ionized water in conical flask by maintaining pH at
25
6.8 and autoclaved at 121OC for 15min. 20ml of media was poured in autoclaved Petri
plates.
Culture of media
For enumeration of Clostridium perfringens, 100μl dilution was taken from third and
fourth dilution (1:1000 and 1:10000 respectively) and spread on RCM media Plates and
incubated in 5% CO2 at 37OC for forty-eight hours. Colonies were counted on electric
Escherichia coli culture
Medium preparation
Eosin Methylene Blue agar (EMB; Lab: 61; Lab M Ltd, Lancashire, United
Kingdom) was utilized for growth E. coli. For this purpose 37.5g of EMB agar was
mixed in one liter of de-ionized water. pH was maintained at 6.8 and autoclaved at 121
OC for 15 minutes. 20 ml media was poured in autoclaved petri plates.
Culturing media
For enumeration of Escherichia coli 100μl dilution was taken from third and
fourth dilution (1:1000 and 1:10000 respectively) and spread on EMB media plates,
incubated aerobically at 37OC for twenty-four hours. E. coli colonies gave green-metallic
sheen on EMB agar and were counted on electric colony counter.
STATISTICAL ANALYSIS
Statistical program SPSS for window (Version 13 SPSS Inc., Chicago, Illinois,
USA) was used for data analysis. Data were expressed as Means ± S.E.M. The S-N-K
test was used to test the normal distribution of the data. The data were analyzed using
one-way analysis of variance. The group differences were compared by the Duncan’s
26
Multiple Range Test (Duncan, 1955). A probability value at P < 0.05 was considered to
be significant.
CHAPTER-4
RESULTS
GROWTH PERFORMANCE
1. Feed Consumption
The results revealed that weekly based and overall feed consumption did not
change in control and MOS supplemented groups (Table 4.1).
2. Body Weight Gain
A significant difference (p<0.05) in the body weight gain of quails was observed
during the first week of age among control and MOS supplemented groups. The body
weight gain was significantly lower in group D compared to control and group B.
However, the body weight gain did not change among control and MOS supplemented
groups during rest of the experimental period. The overall body weight gain was
statistically similar among control and MOS supplemented groups (Table 4.2).
3. Feed Conversion Ratio
A significant difference (p<0.05) in the feed conversion ratio (FCR) was observed
during the first week of age among control and MOS supplemented groups. The FCR of
group D was significantly higher compared to control and group B. However, overall
27
FCR did not change among control and MOS supplemented groups during rest of the
experimental period (Table 4.3).
4. Body Weight
The body weights of the MOS supplemented quails were non-significantly different
compared to the control group. However, body weight of group C was significantly lower
compared to other MOS supplemented groups (Table 4.4).
5. Relative Weights of Visceral Organs
Relative weights of visceral organs of quails have been presented in Table 4.5.
The results revealed that dietary supplementation of MOS did not affect the relative
weights of visceral organs except liver. The relative weight of liver was significantly
higher (p<0.05) in group D compared to control group (Table 4.5).
6. Relative Length of Visceral Organ
The relative length of small intestine with or without digesta of quails has been
presented in Table 4.6. The results revealed that dietary supplementation of MOS did not
significantly affect the relative lengths of small intestine and ceca compared to control
group.
7. Mineral Profile
The results revealed that dietary supplementation of MOS did not affect the
calcium, magnesium, phosphorus, copper and iron concentrations of blood serum (Table
4.7).
8. Microbial Populations of Cecal Digesta
The results revealed that MOS supplementation did not affect microbial
populations of the cecal digesta (Table 4.8).
28
29
TABLE 4.1: Mean feed consumption (g) of control and MOS supplemented groups of quails.TREATEMENT
GROUPS
FEED CONSUMPTION (g)
Week 1 Week 2 Week 3 Week 4 Week 5 Overall
A 33.00 ± 0.77 104.37 ± 4.74 132.62 ± 3.45 145.75 ± 6.93 168.87 ± 6.36 584.62 ± 18.28
B 33.87 ± 1.34 100.75 ± 3.81 135.75 ± 5.57 137.87 ± 4.63 167.87 ± 4.05 576.12 ± 11.98
C 35.37 ± 1.49 97.25 ± 4.76 134.75 ± 4.88 137.12 ± 3.88 171.12 ± 5.37 575.62 ± 12.38
D 36.62 ± 1.36 104.00 ± 4.96 135.00 ± 3.83 141.12 ± 6.17 173.75 ± 6.24 590.50 ± 14.59
Values represent the Mean ± S.E. of four groups of quail chicks.
30
TABLE 4.2: Mean body weight gain (g) of control and MOS supplemented groups of quailsTREATEMENT
GROUPS
BODY WEIGHT GAIN (g)
Week 1 Week 2 Week 3 Week 4 Week 5 Overall
A 12.13 ± 0.18a 32.91 ± 0.84 48.93 ± 0.85 46.73 ± 2.36 43.36 ± 1.98 184.08 ± 1.73ab
B 11.88 ± 0.35a 34.86 ± 0.52 48.86 ± 1.78 44.83 ± 1.39 45.76 ± 1.88 186.21 ± 1.18a
C 11.68 ± 0.31ab 32.20 ± 1.52 51.76 ± 2.03 44.11 ± 1.95 40.30 ± 1.95 180.06 ± 1.78b
D 10.92 ± 0.18b 33.51 ± 0.93 49.22 ± 1.40 49.55 ± 2.23 40.87 ± 2.3 184.08 ± 2.19ab
Values represent the Mean ± S.E. of four groups of quail chicks.a-b Values within columns with no common superscripts are significantly different (P<0.05).
31
TABLE 4.3: Mean feed conversion ratio (FCR) of control and MOS supplemented groups of quails.TREATEMENT
GROUPS
FEED CONVERSION RATIO (g Feed/g BWG)
Week 1 Week 2 Week 3 Week 4 Week 5 Overall
A 2.72 ± 0.05b 3.19 ± 0.21 2.71 ± 0.09 3.16 ± 0.18 3.95 ± 0.22 3.17 ± 0.09
B 2.87 ± 0.16b 2.89 ± 0.12 2.80 ± 0.16 3.10 ± 0.16 3.70 ± 0.16 3.09 ± 0.07
C 3.05 ± 0.17ab 3.04 ± 0.15 2.63 ± 0.14 3.14 ± 0.14 4.35 ± 0.33 3.19 ± 0.06
D 3.37 ± 0.17a 3.11 ± 0.16 2.76 ± 0.14 2.87 ± 0.15 4.37 ± 0.33 3.17 ± 0.07
Values represent the Mean ± S.E. of four groups of quail chicks.a-b Values within columns with no common superscripts are significantly different (P<0.05).
32
TABLE 4.4: Mean body weights (g) of control and MOS supplemented groups of quails.GROUPS INITIAL BODY WEIGHT(g) BODY WEİGHT(g)
A (control) 7.8±0.12 191.25±2.28ab
B (1.0 %-MOS) 7.97±0.10 193.87±1.34a
C (0.5 %- MOS) 7.93±0.09 187.75±1.30b
D (0.1 %-MOS) 8.08±0.18 193.18±1.46a
Values represent the Mean ± S.E. of four groups of quail chicks a-b Values within columns with no common superscripts are significantly different (P<0.05).
33
TABLE 4.5: Mean relative visceral organs weight (g) of control and MOS supplemented groups of quails.
RELATIVE WEIGHTS
(g/g BW)
TREATEMENT GROUPS
A B C D
Small intestine with digesta 3.72 ± 0.16 3.91 ± 0.12 4.09 ± 0.13 3.98 ± 0.15
Small Intestine without digesta 2.45 ± 0.09 2.51 ± 0.06 2.51 ± 0.07 2.49 ± 0.11
Cecum with digesta 0.54 ± 0.04 0.58 ± 0.04 0.56 ± 0.04 0.59 ± 0.04
Cecum with out digesta 0.27 ± 0.01 0.29 ± 0.02 0.30 ± 0.02 0.31 ± 0.01
Gizzard with digesta 3.50 ± 0.15 3.47 ± 0.13 3.49 ± 0.18 3.82 ± 0.22
Gizzard without digesta 2.71 ± 0.12 2.42 ± 0.09 2.44 ± 0.11 2.79 ± 0.16
Heart 0.86 ± 0.03 0.80 ± 0.02 0.82 ± 0.02 0.85 ± 0.03
Liver 2.41 ± 0.11b 2.54 ±0.09ab 2.45 ±0.10ab 2.74 ± 0.11a
Values represent the Mean ± S.E. of four groups of quail chicks.a-b Values within rows with no common superscripts are significantly different (P<0.05).Group A (control) Group B (1% MOS), Group C (0.5% MOS) or Group D (0.1% MOS). BW=body weight.
34
TABLE 4.6: Mean relative visceral organs length (cm) of control and MOS supplemented groups of quails.
RELATIVE LENGTHS
(cm/g BW)
TREATEMENT GROUPS
A B C D
Small intestine with digesta 30.32 ± 0.63 29.93 ± 0.75 31.12 ± 0.65 30.69 ± 0.60
Small Intestine without
digesta 32.34 ± 0.78 31.37 ± 0.79 32.79 ± 0.63 32.11 ± 0.62
Values represent the Mean ± S.E. of four groups of quail chicks. Group A (control) Group B (1% MOS), Group C (0.5% MOS) or Group D (0.1% MOS). BW=body weight
35
TABLE 4.7: Mean serum mineral concentrations of control and MOS supplemented groups of quails.ATTRIBUTES TREATEMENT GROUPS
A B C D
Ca (mg/dl) 10.33 ± 0.83 9.32 ± 0.91 9.80 ± 1.29 11.39 ± 1.11
P (mg/dl) 4.57 ± 0.39 5.40 ± 0.12 4.77 ± 0.44 5.22 ± 0.28
Mg (mg/dl) 3.81 ± 0.32 3.46 ± 0.19 3.27 ± 0.30 3.92 ± 0.34
Fe (ppm) 3.28 ± 0.17 2.80 ± 0.15 3.16 ± 0.16 2.91 ± 0.18
Cu (ppm) 0.38 ± 0.03 0.36 ± 0.02 0.39 ± 0.03 0.35 ± 0.02
Values represent the Mean ± S.E. of four groups of quail chicks. Group A (control) Group B (1% MOS), Group C (0.5% MOS) or Group D (0.1% MOS).
36
TABLE 4.8: Mean change in cecal digesta of control and MOS supplemented groups of quails.
ATTRIBUTES TREATEMENT GROUPS
A B C D
Escheria coli( x 106 c.f.u /g
cecal contents)
4.54 ± 1.03 3.98 ± 1.03 4.54 ± 1.03 4.88 ± 0.97
Clostridium Perfringens (x103
c.f.u / g cecal contents)
2.34 ± 0.60 2.33 ± 0.60 2.02 ± 0.59 1.82 ± 0.60
Group A (control) Group B (1% MOS), Group C (0.5% MOS) or Group D (0.1% MOS);
37
CHAPTER-5
DISCUSSION
Prebiotics are the carbohydrates which are not digested by the digestive enzymes
of the host and are fermented by the beneficial intestinal bacteria and thus are beneficial
for host (Gibson and Roberfroid, 1995). Mannan-oligosaccharide (MOS), a prebiotic is
derived from the cell wall of Saccharomyces cerevisiae and is commercially available as
a feed supplement included in diets as a beneficial compound. The benefits of MOS are
based on specific properties that include modification of the intestinal microflora,
reduction in turnover rate of the intestinal mucosa and modulation of the immune system
in the intestinal lumen. These properties have the potential to enhance growth rate, feed
efficiency, egg production and livability in poultry species (Shane, 1999). Prebiotics have
been shown to improve body weight gain and feed conversion efficiency in turkeys (Sims
et al., 2004; Fritts and Waldrop, 2003) and broilers (Hooge et al., 2003).
Feed Consumption
The results of the present study revealed that the MOS supplementation did not
affect feed consumption. Similar findings have been reported in the broilers (Midilli et
al., 2008; Cakir et al., 2008; Jung et al., 2008) and in quails (Parlat et al., 2003; Ghosh et
al., 2007).These results did not support the findings of Eleftherios et al. (2010) and Oguz
and Parlat (2004) who reported that in quails, feed consumption increased significantly.
In contrast, Rosen (2007a; 2007b) in two comparative studies reported lower feed
consumption for birds fed MOS versus controls. It is considered that due to the presence
38
of Mannan-oligosaccharides, undesirable microorganisms from the gastrointestinal tract
are eliminated that results in reduction of the stress on the mucosa caused by pathogens
and dietary nutrients are absorbed in a normal way.
Body Weight Gain
In the present study a significant difference (p<0.05) in the body weight gain of
quails was observed after the first week of age among control and MOS supplemented
groups, whereas, the overall body weight gain was statistically similar among control and
MOS supplemented groups. Similar results were reported by Ammerman (1989) and
Waldroup et al. (1993). Flemming et al. (2004) also found that MOS has improved the
BWG in broilers. Yang et al. (2007) studied the effects of Mannan-oligosaccharide on
growth performance of broiler and reported that there were no significant differences in
BWG among treatments; however, the MOS supplementation tended to improve BWG in
early life of broiler chicks.. In contrast with present results, Ghosh et al. (2007) reported
that MOS supplementation did not increase body weight gain in quail. It is considered
that due to the presence of Mannan-oligosaccharides; pathogens from the gastrointestinal
tract are eliminated that results in reduction of the stress on the mucosa caused by
pathogens. Thus absorption and utilization of the dietary nutrients increased that result in
higher body weight gain.
Feed Conversion Ratio
In the present study, a significant difference in the feed conversion ratio (FCR)
was observed during the first week of age among control and MOS supplemented groups,
however, during rest of the experimental period, FCR remained unchanged. Similar
results were reported by Eleftherios et al. (2010), who reported that the addition of MOS
39
resulted in a tendency for higher FCR in Japanese quails during the fourth week which
was not significant. These results are in contrast with Midilli et al. (2008) who found that
inclusion of prebiotics improved the feed conversion ratio in broilers. Parlat et al. (2003)
reported that feeding MOS improved overall feed conversion ratio for 0-5 wks of age in
Japanese quails. This improvement in FCR is in agreement with the findings of Parks et
al. (2001), who found that MOS-supplemented diets showed a lower FCR of the birds.
Similarly, Guclu (2003) and Ghosh et al. (2007) found lower FCR for birds fed MOS.
Hooge (2004) based on a meta-analysis of 24 broiler pen trials, reported that Bio-Mos
decreased FCR by an average of 1.99% compared with the control group. Similarly,
Rosen (2007) from statistical evaluation of 82 comparisons with negative control diets
found that Bio-Mos diets reduced FCR by 0.039. Savage and Zakrzewska (1997) in
turkeys and Waldroup et al. (2003) in broilers reported that MOS supplementation
improved FCR significantly, Whereas, Yalqnkaya et al. (2008) found that Mannan-
oligosaccharides did not affect the feed conversion ratio in broiler.
Body Weight
In the present study, body weights of the MOS supplemented quails were non-
significantly different compared to the control group. In contrast to present findings,
Parlat et al. (2003); Oguz and Parlat (2004); Guclu (2003) and Eleftherios et al. (2010) in
quails and Frittis and Waldroup (2003) and Parks et al. (2001) in turkeys observed higher
body weight in birds that consumed MOS. Flemming et al. (2004); Hooge (2004) and
Bentes et al. (2008) found the same results in broilers, whereas, Ghosh et al. (2007) and
Sarica et al. (2009) did not observe any significant difference in body weight among
MOS supplemented and control groups in quails.
40
Relative Weights and Lengths of Visceral Organ
Results of the present study revealed that dietary supplementation of MOS did not
affect the relative weights of heart, gizzard, cecum and small intestine (both filled and
empty), whereas, relative weight of liver was significantly higher in MOS supplemented
group D compared to control group. The results of present study are similar to other
studies done by Mohamed et al. (2008) who found that dietary MOS has no effect on the
relative weights of heart and gizzard of broilers. Similar results were found in the study
of Rehman et al. (2007a,b), who found that inuline did not affect the relative weight and
length of the small intestine in broilers. In present study, the results of the relative weight
of liver are in contrast with the findings of Mohamed et al. (2008) who reported that
MOS supplementation did not significantly affect relative weight of liver in broilers.
Similarly, Rehman et al. (2007a,b) also reported that feed composition did not affect the
weight of liver in broilers. Whereas, Eleftherios et al. (2010) reported that MOS
supplementation decreased the liver to live weight percentage in quails. Results of the
relative length of small intestine are similar with the study of Juskiewicz et al. (2002) and
(2004) who reported that different oligosaccharides have no effects on the length or
weight of the small intestine in turkeys.
The growth promoting effect of MOS is due to their ability to limit the growth of
pathogens in the digestive tract of animals (Bozkurt et al., 2008) and increase the
population of useful bacteria (Ghosh et al., 2007). Thus, the digestive tract remains
healthy, functions more efficiently and more nutrients are absorbed. The lack of
significant improvement in the performance of the birds that was found in our experiment
may be the result of the proper feed composition and the optimum rearing conditions. It is
41
generally accepted that the positive effect of feed growth promoters are more obvious
when animals are not offered good quality feed or are reared in non-optimum conditions
or even bids are kept in un-hygienic or in stress conditions (Sims et al., 2004; Baurhoo et
al., 2007; Bozkurt et al., 2008). Ferket (2004) relates that the best performance of the
birds fed with Mannan-oligosaccharides (MOS) diets is due to the:
Increase of the resistance to the intestinal pathogenic microorganisms.
Reduction of the competition between bacteria and host for the starch and sugars.
Changing intestinal pH, that ultimately suppresses the explosion of pathogenic
bacteria.
Serum Minerals
In the present study, dietary supplementation of MOS did not affect the calcium,
magnesium, phosphorus, copper and iron concentrations of blood serum. These results
support the findings of Van den Heuvel et al. (1998); Ellegard et al. (1997); Tahiri et al.
(2003) and Lopez-Huertas et al. (2006) who reported that prebiotics like inulin,
oligofructose, or other nondigestible oligosaccharide (NDO) did not affect calcium or
iron absorption in humans. These results are in contrast with findings of Abrams et al.
(2005); Griffin et al. (2002); Van den Heuvel et al. (1999) who found that prebiotics
stimulated the absorption of calcium in humans. Ghosh et al. (2008) also found that MOS
treated groups exhibited significantly higher Ca compared to control group in Japanese
quail. Similarly, Scholz-Ahrens et al. (2001) also reported that inuline stimulate
absorption of several minerals, particularly calcium and magnesium in adult rats. Similar
findings were reported by Coudray et al. (1997) in healthy human adults and added that
inulin increased calcium absorption and had no effect on the metabolism of the other
42
minerals like magnesium, phosphorous, copper and iron. Reporting of conflicting results
regarding mineral absorption may be due to the experimental design because the effect of
NDOs depends on the dose, the time of administration, the content of calcium in the diet,
and the age of the subjects studied.
Mechanism
The mechanism involved numerous factors that contribute in mineral absorption
which includes:
Increased bacterial production of short-chain fatty acids through increased supply
with substrate.
Enlargement of the absorption surface by promoting proliferation of enterocytes
mediated by bacterial fermentation products, chiefly lactate and butyrate.
Increased expression of calcium binding proteins.
(Scholz-Ahrens et al., 2001 and 2002; Coudray et al., 2003; Cashman et al., 2003)
Microbial Populations of Cecal Digesta
The results of the present study revealed that MOS supplementation did not affect
Escherichia coli, and Clostridium perfringens populations of the cecal digesta. These
results favour the findings of Yang et al. (2007), who reported that Clostridium
perfringens were not affected in broilers by the supplementation of MOS. Similar
findings were also reported by Spring et al. (2000) and Ceylan et al. (2003) who reported
that Mannan-oligosaccharide did not significantly reduce the concentrations of cecal
pathogens in broilers. Finucane et al., (1999) also reported that there was no significant
difference in the level of Clostridium spp. in cecal contents of turkeys.
43
The results of the present study are in contrast with some of the findings of the
previous studies stating that MOS also resulted in a major reduction in Escherichia coli.
Baurhoo et al. (2007) reported that reduction of the cecal concentration of total
Escherichia coli due to MOS supplementation was more pronounced in Escherichia coli -
challenged birds. Fairchild et al. (2001) also reported the similar results and added that
MOS provides protection to chicks by reducing some of the pathogenic bacteria such as
Escherichia coli. Young et al. (2008) also reported that dietary MOS also reduced the
counts of Clostridium perfringens in the ceca of birds. However, Brzoska et al. (2005)
reported that birds receiving MOS had more Escherichia coli compared with the
antibiotic treatment.
44
CHAPTER-6
SUMMARY
Long term use of antibiotics has side effects like antibiotic resistance and drug
residues in meat that ultimately harm the humens. To avoid such hazards, it was
necessary to find out an alternative of antibiotics. Prebiotics are considered as an
alternative to antibiotics as prophylactic, therapeutic and growth-promoting agents in
poultry production. Keeping in view the present scenario; a 35 day long feeding trial was
conducted.
A total of 1320 day old Japanese quail chicks were randomly divided into 4
groups (n=320) with 8 replicates (n=40). Birds were fed a corn-based basal diet (Group
A) or the same basal diet supplemented with MOS either at 1% (Group B), or 0.5%
(Group C) or 0.1% (Group D) levels. Feed consumption, body weight gain, FCR, total
body weight, relative weights and lengths of visceral organs, serum calcium, magnesium,
phosphorus, copper and iron and microbial population of ceca were the parameters
considered.
Results showed that body weight gain, FCR and relative weight of liver of the
MOS supplemented quails were significantly (p<0.05) different and Final body weights
were non-significantly (p>0.05) different compared to control group. Whereas, feed
consumption, relative weights of other visceral organs, serum calcium, magnesium,
phosphorus, copper & iron and cecal microbial populations were not influenced by
treatments.
CHAPTER-7
45
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Ammerman, E., C. Quarles and P. V. Twining Jr. (1989).Evaluation of fructo-
oligosaccharides on performance and carcass yield of male broilers. Poult. Sci.
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AOAC. (1995).Official method of the association of official analytical chemists. Vol. 1,
6th Ed. AOAC. International, Arlington, USA.
Abrams, S. A., I. J. Griffin, K. M. Hawthorne, L. Liang, S. K. Gunn, G. Darlington and
K. J. Ellis (2005).A combination of prebiotic short- and long-chain inulin-type
fructans enhances calcium absorption and bone mineralization in young
adolescents. Am. J. Clin. Nutr.82:471–476.
Bio-Mos Product Specifications (1997). Bio-Mos Poultry Dossier, Alltech Inc.,
Nicholasville, KY.
Bauer, E., B. A.William, M. W. A. Verstegen and R. Mosenthin (2006). Fermentable
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60
VITA
MUHAMMAD ANWAR IQBAL
PERMANENT ADDRESS:
Ward # 6 Chowk Azam, Tehsil & District Layyah.
E.Mail: anwariqbalk@yahoo.com
Contact No: 0300 8121182
EDUCATION
Master of Philosophy: PHYSIOLOGY, UVAS, Lahore. June, 2010.
Dissertation Title: Effect of Dietary Supplementation of Mannan-Oligosaccharides on
Growth Performance, Cecal Microbial Population and Mineral Absorption in Japanese
Quail (Coturnix Coturnix Japonica).
Advisor: Dr. Habib Rehman, Ph.D.
Master of Science: ZOOLOGY, BZU, Multan. June, 2004.
Thesis Title: Effects of Exercise on Lipid profile on healthy volunteers of Baha-u-ddin
Zakaryya University Multan.
Advisor: Dr. Tassawar Hussain Khan, Ph.D.
Bachelor of Science: ANIMAL SCIENCES, BZU, Multan. September, 2001.
61
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