Abstract: An experiment was conducted to determine the influence of organic acid salts and mannan oligosaccharides 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 10 chicks per replicate. Control (C0) birds were given a standard basal diet, C1 birds were provided with basal diet having antibiotic (Bacitracin methyl disalisylate-BMD) at 0.5 g kg-1; T1 birds having diet with sodium butyrate at 5 g kg-1; T2 birds were provided with basal diet having Mannan Oligosaccharide (MOS) at 1 g kg-1; group T3 birds were fed basal diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1 and sodium butyrate 5 g kg-1; T4 birds having diet with sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1; T5 birds having diet with ammonium formate at 1 g kg-1, calcium propionate at 1 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg -1 of the diet. Organic acid salt supplementation increased (p<0.05) live weight, live weight gain compared to control (C). Cumulative feed intake was not significantly affected due to dietary treatments. Superior results in terms of Feed Conversion Ratio (FCR) and Performance Index (PI) were found in organic acid salt combination (T3) and organic acid salts combination (OASC) with MOS (T5) supplemented groups compared to others. OASC and OASC with MOS improved (p<0.05) gut health by reducing bacterial load in gut compared to control (C0) and other groups also. The result showed OASC and OASC with MOS supplementation significantly improved villus height compared to other groups. This experiment revealed that the use of OASC or OASC with MOS in the diet improved body weight gain, FCR, PI, villus height and reduced bacterial load in gut as antibiotic compared.
INTRODUCTION
Public disapproval for the use of antibiotics in the poultry production due to their residual effects in meat, concerns over the emergence of antibiotic resistant Superbugs (JETACAR, 1999). The problem is however much more complicated that just a simple withdrawal of this growth promoters overnight, it will not only affect feed efficiency but also increase mortality and morbidity of animals. Therefore, many parts of the world are experimenting alternative feed additives that may be used to alleviate the problems, associated with the withdrawal of antibiotic from feed. In poultry production organic acids like formic and propionic have mainly been used in order to sanitize the feed having problem with bacterial infection (Hinton and Linton, 1985; Berchieri and Barrow, 1996; Thompson and Hinton, 1997). Butyrate is considered to be responsible for normal development of epithelial cell (Pryde et al., 2002). Mannan oligosaccharides (MOS), derived from yeast cell wall, maintain gut health by adsorption of pathogenic bacteria containing type-I fimbrae of different bacterial strains and remove the bacteria from gut (Oyofo et al., 1989; Spring et al., 2000) and increase villus height, uniformity and integrity (Loddi et al., 2002). Under these circumstances, the present study was carried out to evaluate the efficiency of various organic acid salts, viz., formic acid, propionic acid and butyric acid salt, as well as MOS as growth promoter and performance enhancer of Japanese quail.
MATERIALS AND METHODS
Day old chicks of Japanese quail (n = 280) were randomly distributed into seven experimental groups consisting of four replicates in each. Each replicate was with 10 birds. The chicks were individually weighed and allocated to 28 cages of electrically heated battery brooder so that each cage had birds with similar (p>0.05) average weight. In control (C0) birds were supplied with standard basal diet; group C1 birds were provided with diet having antibiotic Bacitracin methyl disalisylate (BMD) at 0.5 g kg-1 of diet; group T1 birds having diet with sodium butyrate at 5 g kg-1 diet; group T2 birds were provided with diet having MOS 1 g kg-1 diet; group T3 birds were fed diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1 and sodium butyrate 5 g kg-1 of the diet; group T4 birds having diet with sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of the diet; group T5 birds having diet with ammonium formate at 1 g kg-1, calcium propionate at 1 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of the diet. Basal diet was starter (0 to 3 weeks) and finisher (4 to 6 weeks). The diets (Table 1) of the experiment were formulated to meet or exceed the nutrient requirement as per Bureau of Indian Standard (BIS, 1992) .
Housing and Management
Chicks were housed in a electrically heated battery brooder which were disinfected
thoroughly three days prior to arrival of birds. The day-old chicks were offered
electrolyte solution upon arrival. Birds were maintained on a 24 h constant
light schedule. Body weight was recorded at the initial day followed by at weekly
interval up to 6 weeks before offering feed. FCR, feed intake and live weight
of birds were recorded at weekly intervals.
| Table 1: | Composition of basal diets for starter and finisher for quail (By parts) |
| +: Composition of trace minerals. Manganese Sulphate 15 g, Copper Sulphate 2 g, Ferrus Sulphate 6 g and Zinc Carbonate 6 g. ++: Composition of DAILYMIX. Vitamin A 82,500 IU, Vitamin B2 50 mg, Vitamin D3 12,000 IU, Vitamin K 10 mg, Vitamin B12 15 mcg. +++ Composition of BRIPLEX. Vitamin B1 8 mg, Vitamin B2 4 mg, Vitamin B6 16 mg, Vitamin B12 80 mcg, Niacin 120 mg, Calcium Pantothenate 80 mg, Folic acid 3600 mcg and Vitamin E 80 mg | |
PI for each group was calculated from total body weight gain and feed efficiency ratio.
Dietary Ingredients of Feeds
The dietary ingredients were in Table 1. Total protein was analyzed according
to AOAC (1995).
Measurement of pH of Different Parts Gastrointestinal (GI) Tract
To determine the pH of gut of birds, 10 g of gut content from crop, proventriculus,
gizzard, duodenum, jejunum and ileum were collected aseptically in 90 mL sterilized
physiological saline (1: 10 dilution) (Al-Natour and Alshawabkeh, 2005) and
pH was determined.
Measurement of Villus Height
For measurement of villus height, parts of small intestine were taken during
slaughter. Then slides were prepared by sectioning and staining of tissues from
parts of intestine. Heights of intestinal villus were measured by ocular micrometer
under 10x objective of microscope. The reading was taken from ocular micrometer
and the actual villus height was obtained by multiplying the ocular micrometer
reading by conversion factor derived from stage micrometer (Lillie, 1965). The
heights of intestinal villus were expressed as micrometer (μm).
Microbial Count in Intestinal Content
Two birds from each replicate of different group were electrically stunned
and sacrificed at 6th week of age. Intestine including duodenum, jejunum, ileum
were removed and ligated at both sides and placed into 50 mL tubes in normal
saline and then kept at 4°C until used for intestinal sampling. Serial dilution
of collected samples from different parts of intestinal contents were made upto
fifth dilution with normal saline and different bacterial loads of content were
enumerated by pour plate method (Quinn et al., 1992).
Statistical Analysis
The analysis was done by using General Linear Model of SPSS programme (SPSS,
1997) with replicates as experiment units for studying the effect of organic
acid on broiler performance and gut health. Levels of significance were calculated
by Duncan Test (Duncan, 1995) whenever any effect was found significant.
RESULTS AND DISCUSSION
Effect on Performance
0-3 Weeks
The average body weight and cumulative feed intake (Table 2) of Japanese
quail upto 3 weeks were not varied (p>0.05) due to dietary treatments but
significant variation was found only in FCR and Performance Index (PI). Combination
of acid salts with MOS (T5) showed highest (p<0.05) PI over other
groups. Group C1, T3 and T1 were also comparable
with T5. Even group C0 showed better performance index
than MOS supplemented group. FCR was not affected (p>0.05) due to supplementation
of OAS or in combination form of OAS and MOS when compared to control. However,
supplementation of individual MOS in the diet (T3) increased (p<0.05)
FCR.
3-6 Weeks
It was observed that supplementation of only acidifier combination and acidifier
combination with MOS showed better (p<0.01) body weight gain, FCR and PI
(Table 2) compared to others. No difference (p>0.05) in
body weight gain was seen in control (C0) and MOS (T2)
supplemented group.
| Table 2: | Effect of feeding organic acid salts and MOS on the performance traits of Japanese quail† |
| † Values bearing no common superscript in a row differs significantly (p<0.05). C0 birds supplied with basal diet; C1, T1 and T2 provided with diet having BMD at 0.5 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of diet, respectively. T3 birds were fed diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1 and sodium butyrate 5 g kg-1, T4 birds having diet with sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 and T5 birds having diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of the diet | |
0-6 Weeks
Average live body weight (Table 2) after 6 weeks varied due to dietary treatments
showing highest (p<0.05) body weight in acidifier combination with MOS supplemented
group followed by T3 T4, T1 and C0.
Control group attained lowest (p<0.05)) body weight than all other treatment
groups. FCR and PI were found better (p<0.01) in acidifier combination and
acidifier combination with MOS supplemented group compared to others. Individual
MOS supplemented group showed significantly better FCR than control group (C0).
In some of the previous studies it was found that organic acids as a substitute
of AGPs improved the performance of birds even in absence of antibiotic (Mairoka
et al., 2004). Benedetto (2003) also observed mixture of organic acids
can replace Antibiotic Growth Promoters (AGPs) and acid salts improved production
performance along with other beneficial effects. Present study also showed that
T3 group (combination of acid salts) attained better performance
in live weight gain, FCR and PI compared to others groups except T5
Stanley et al. (2000) found that addition of 0.1% MOS in the Tom’s
diet significantly increase the body weight gain than negative control. Similarly
Parks et al. (2001) reported from a study in turkey supplemented with MOS, utilized
as alternative to AGPs to improve turkey performance. In this experiment MOS
supplemented result, partially support the earlier studies. Live weight gain
of Japanese quail in MOS supplemented group (T3) after 6 week of
age was comparable to antibiotic but no significance difference with control
group (C0). On the other hand FCR and PI of T3 group had
significant difference with control group. Data on live weight gain showed any
marked difference with MOS in the diet of Japanese quail which complies with
the report of Elangovan et al. (2005). Vander Wielen et al. (2000)
reported birds previously fed butyrate can better withstand the stress of coccidial
challenge. In the present study Japanese quail received butyrate showed better
result in live weight gain, FCR and PI than control as well as consistent result
with antibiotic. Butyrate + MOS and mix of acid salts + MOS gave good result
in live weight gain. OASC + MOS attained highest body weight gain after 6 weeks
period with best FCR and PI. Organic acids in un-dissociated form kill the pathogenic
bacteria (Nursey, 1977) and on the other hand MOS being undigested in GI tract
act as a ligand, bind with the type I fimbrae of pathogenic bacteria and thus
increase the population of helpful bacteria in the gut.
| Table 3: | Effect of Organic acid salts and MOS on the gut microbial count of Japanese quail† |
| † Values bearing no common superscript in a row differs significantly (p<0.05). C0 birds supplied with basal diet, C1, T1 and T2 provided with diet having BMD at 0.5 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of diet, respectively. T3 birds were fed diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1 and sodium butyrate 5 g kg-1, T4 birds having diet with sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 and T5 birds having diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of the diet | |
So it may be inferred that the synergistic effect of OASC with MOS resulted superior performance in Japanese quail.
Effect on Gut Microbial Count
The intestinal content of experimental Japanese quail was subjected to gut
microbial count like Coliform, Escherichia coli (E. coli) and
Clostridium perfringens (Cl. perfringens) were counted and presented
in Table 3. It was seen that acidifier combination with MOS supplementation
reduced (p<0.01) coliform count followed by T3, T4,
T2, C1, T1 and C0. The
coliform count in C1, T2, T3 and
T5 group showed significant difference (p<0.05) with control (C0)
group. In case of E. coli the treatment group T5 (acidifier
combination + MOS) possessed lowest bacterial count followed by T4,
T3, T2, C1, T1 and C0.
AGP supplementation in diet resulted reduced (p<0.05) Cl. perfringens
in the intestine. However, dietary treatment with OASC and MOS resulted
to reduced (p<0.05) Cl. perfringens than control group (T0).
Here MOS and organic acid salts supplementation were comparable with AGP group
(T1). From the trend of the analysis it was indicated that MOS and
organic acid salts supplementation groups had an influence to reduce coliforms,
E. coli and Clostridium in the gut of experimental birds which
was in accordance with the findings of Lon (1995), Spring et al. (2000),
Fairchild et al. (2001) and Sims et al. (2004).
The reduced microorganism in the G.I. tract of Japanese quail with the treatment of MOS might be due to block type-I fimbriae which prevent pathogens from attaching to the intestinal lining and passes them out of gut (Dawson and Pirvulescu, 1999). On the other hand organic acids in their undissociated forms pass through the cell membrane of the bacteria and dissociate to from H+ ions which lower the pH of bacterial cell, causing the organism to use its energy, trying to restore the normal balance. Whereas Rcoo- anions produced from the acid can disrupts DNA, hampering protein synthesis and putting the organism in stress. As a result the organism can not multiply rapidly and decrease in number (Nursey, 1997).
Effect on Villus Height
The result showed that only OASC and OASC with MOS supplementation significantly
improved villus height of duodenum, jejunum, ileum (Table 4)
compared to other groups. However, the villus height of ileum did not significantly
differ in acidifier combination and butyrate+ MOS supplementation group. Earlier
workers Macari and Mairoka (2000) and Pelicano et al. (2005) reported
higher villus height in small intestine with most organic acidifier and MOS
in diet of broiler. The increase of villus height of different segment of small
intestine may be attributed due to intestinal epithelium acts a natural barrier
against pathogenic bacteria and toxic substances that are present in the intestinal
lumen.
| Table 4: | Effect of organic acid salts and MOS on the villus height in different parts of intestine of Japanese quail† |
| † Values bearing no common superscript in a row differs significantly (p<0.05). C0 birds supplied with basal diet; C1, T1 and T2 provided with diet having BMD at 0.5 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of diet, respectively. T3 birds were fed diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1 and sodium butyrate 5 g kg-1, T4 birds having diet with sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 and T5 birds having diet with ammonium formate at 1 g kg-1, calcium propionate 1 g kg-1, sodium butyrate at 5 g kg-1 and MOS at 1 g kg-1 of the diet | |
Organic acidifier may reduce the growth of many pathogenic or non-pathogenic intestinal bacteria, therefore, reduce intestinal colonization and infectious process, ultimately decrease inflammatory process at the intestinal mucosa resulting improved villus height and function of secretion, digestion and absorption of nutrients (Iji and Tivey, 1998; Pelicano et al., 2005).
CONCLUSION
From the present study it may be inferred that the organic acid combination along with MOS have better performance compared to antibiotic supplementation in the diet. The improved villus height in duodenum, jejunum and ileum in the respective group also supports the finding. Thus, it may be concluded that organic acid combination along with MOS can be a potential tool as an alternative to antibiotic in the diet of Japanese Quail for it’s optimum production.