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

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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

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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

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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.

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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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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