Abstract

Six hundred male broilers were divided into two groups: group I, fed on a conventional diet, and group II, fed on a diet supplemented with probiotics. Group II was orally administered a combination of three probiotics (Bacillus mesentericus TO-A, Clostridium butyricum TO-A and Streptococcus faecalis T-110). Growth performance, gut mucosal immunity, and carcass and meat quality of broilers in both groups were assessed after being reared for 49 days. At 9, 14, 34 and 49 days, birds in group II showed a significantly greater body weight (p <0.05) than birds in group I (control). Average daily gain for 0–9, 0–14, 0–34 and 0–49 days was significantly (p <0.05) higher for birds in group II than that for birds in group I. For the entire period, the feed conversion ratio in group II was significantly lower (p <0.05) than that in group I. At 28 and 49 days, birds in group II had significantly lower serum total cholesterol and triglyceride concentration (p <0.05) than birds in group I. Group II had significantly higher percentages of carcass yield than those in group I. In addition, group I had lower percentages of abdominal fat than those in group II (p <0.05). In leg and breast meat, group II had significantly lower percentages of fat than group I (p <0.05). At 49 days, birds in group II had significantly greater jejunum and ileum IgA concentration than birds in group I (p <0.05). It is suggested that supplementation of broiler diet with probiotics improved growth performance, gut mucosal immunity and carcass and intramuscular fat.

Keywords: growth performance, gut mucosal immunity, carcass and meat quality, broilers, probiotics

Author and Article Information

Affiliation1 Toa Pharmaceutical Co., Ltd.

RecievedJun 18 2015 Accepted: Oct 05 2015 Published: Oct 21 2015

CitationInatomi T (2015) Growth performance, gut mucosal immunity and carcass and intramuscular fat of broilers fed diets containing a combination of three probiotics. Science Postprint 1(2): e00052. doi: 10.14340/spp.2015.10A0001.

Copyright©2014 The Authors. Science Postprint published by General Healthcare Inc. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 2.1 Japan (CC BY-NC-ND 2.1 JP) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

Funding None

Competing interest None

Ethics Ethical approval was obtained from the ethics committee of Toa Pharmaceutical Co., Ltd.

Corresponding author: Takio Inatomi

Address: Toa Pharmaceutical Co., Ltd. 2-1-11, Sasazuka, Shibuya, Tokyo 151-0073, Japan

E-mail: takato.inatomi@gmail.com

Peer reviewerRyuichi Tatsumi¹ and Reviewer B
1 Department of Animal and Marine Bioresource Sciences, Graduate School of Agriculture, Kyushu University

Introduction

The use of antimicrobial growth promoters in animal nutrition has been beneficial for the improvement of growth performance and prevention of diseases 1. Recently, biosecurity threats caused by the resistance of pathogens to antibiotics and accumulation of antibiotic residues in animal products and the consumer 2 required a limitation on the global use of antimicrobial growth promoters. To respond to market demands, the poultry industry is actively searching for alternatives to antibiotics that can both maintain performance levels and be economically viable. A variety of functional preparation such as herb, essential oil and organic acid has been tried. Probiotics provide a possible solution from the viewpoint of multiple beneficial effects. Probiotics are live organisms that favourably affect the animal body when consistently provided in the diet and act to balance the intestinal flora in a symbiotic relationship, which positively influences intestinal villi 3. As a consequence, digestion and absorption of nutrients can be improved. Multiple beneficial effects of including probiotics in the diet of broilers have been reported, including 1) performance 4–7; 2) nutrient digestibility 8, 9; 3) modulation of intestinal microflora 7, 10–12; 4) inhibition of pathogens 13–15; 5) immunomodulation and gut mucosal immunity 4, 10, 11 and 5) improved carcass yield and meat quality 16, 17. However, to the best of our knowledge, there are very few studies reporting on the use of a combination of three probiotics in broiler chickens.

Therefore, this study was undertaken to determine the effect of supplementing broiler diet with a combination of three probiotics on growth performance, gut mucosal immunity and carcass and intramuscular fat in broilers.

Materials and Methods

Ethical approval

This study was conducted at the commercial poultry farm in Kagoshima prefecture in Japan and was conducted according to the fundamental guidelines for proper conduct of animal experiments and related activities in academic research institutions under the jurisdiction of the Ministry of Education, Culture, Sports, Science and Technology.

Ethical approval was obtained from the ethics committee of Toa Pharmaceutical Co., Ltd.

Birds, diets and experimental design

A total of 600 Cobb500™ male broiler chicks at 1 day of age were weighed and divided into two groups, with group I fed a conventional diet and group II fed on a diet supplemented with probiotics, and both groups were reared to the end of the experimental study at 49 days. A standard management procedure was used throughout the experiment. All birds were provided feed and drinking water ad libitum and were offered identical antibiotic-free basal diets. A probiotic cocktail (AVITEAM, Toa Pharmaceutical Co. Ltd., Tokyo, Japan) containing Bacillus mesentericus TO-A at 1 x 10⁸ CFU g⁻¹, Clostridium butyricum TO-A at 1 x 10⁸ CFU g⁻¹ and Streptococcus faecalis T-110 at 1 x 10⁹ CFU g⁻¹ in corn starch was supplemented to the feed at 0.02% (w/w). Commercially available poultry feed (Kagoshima Agricultural Economic Federation) was used as the formulated diet throughout the experimental study. The broiler chicks were fed with standard broiler starter at 1–9 days (CP: 21% ME: 2,988 kcal kg⁻¹), broiler grower A at 10–14 days (CP: 19% ME: 3,083 kcal kg⁻¹), broiler grower B at 15–34 days (CP: 18% ME: 3,176 kcal kg⁻¹) and broiler finisher at 35–49 days (CP: 17% ME: 3,176 kcal kg⁻¹) rations as formulated by the Kagoshima Agricultural Economic Federation. The composition of the diets was shown in Table 1. The birds were reared under hygienic management practises throughout the entire period of study.

Growth performance

Chickens were weighed individually on 1, 9, 14, 34 and 49 days to determine body weight and average daily gain. Feed intake was weighed at 1–9, 10–14, 15–34 and 35–49 days, and feed conversion rate (feed consumed per weight gain) was also calculated.

Serum total cholesterol and triglyceride

At 28 and 49 days, 50 birds from each group were randomly chosen and blood samples (2 ml) were taken from the jugular vein. Serum was isolated by centrifugation at 3,000 xg for 20 min and stored at −70°C until analysis. Serum cholesterol and triglyceride concentrations were measured using a blood automatic analyser (HITACHI 7060, Tokyo, Japan).

Carcass yield, abdominal fat and proximate composition

At the end of the experiment (49 days of age), birds were slaughtered to evaluate yield. Fifty birds from each group were randomly chosen, identified, individually weighed and fasted for 6 h with water provided ad libitum. Carcasses were weighed after removing the feet, head and neck. Abdominal fat (%) and proximate composition of cuts (leg and breast) were calculated. Moisture content was determined by the oven method (AOAC, 1999; 18) and protein content by the Kjeldahl method (AOAC, 1999; 18). Fat composition was determined by the Soxhlet apparatus method (AOAC, 1999; 18) and ash composition was determined using a furnace at 600°C (AOAC, 1999; 18).

Jejunum and ileum IgA concentration

Jejunum and ileum samples were collected randomly from fifty birds from each group at 49 days old. Jejunum and ileum samples were extracted after the birds were killed by cervical dislocation. The jejunum and ileum were excised from each bird. Sample tissues were separated and stored at −20°C until analysis. For analysis, samples were thawed to room temperature. Deionised water was added to 2g of each sample, and it was homogenised for 30 s using a mechanical homogeniser (VirTis, Gardiner, NY). An aliquot (5 ml) of the sample was centrifuged at 20,000 xg for 30 min. The supernatant was collected and stored at −20°C until analysis for IgA concentration. Jejunum and ileum IgA concentrations were measured by chicken-specific IgA ELISA quantitation kits (Jiancheng Biological Engineering Research Institute, Nanjing, China). The ELISA procedure was conducted according to the protocol of the manufacturer, and absorbance was measured at 450 nm.

Statistical analysis

A student’s t-test was performed using the EZR software (Saitama Medical Center, Jichi Medical University). EZR is a graphical user interface for R (The R Foundation for Statistical Computing, version 2.13.0). Differences among means of the dietary treatment groups were compared using least significant differences. A significance level of p <0.05 was used.

Results

Growth performance

The body weight, average daily gain and feed conversion ratio of each group are shown in Tables 2, 3 and 4.

At 9, 14, 34 and 49 days old, birds in group II had significantly greater body weight than birds in group I (p <0.05). The average daily gain for 0–9, 0–14, 0–34 and 0–49 days was significantly higher in group II than that in group I (p <0.05). For the entire period (7 weeks), the feed conversion ratio in group II was significantly lower than that in group I (p <0.05).

Serum total cholesterol and triglyceride

The serum total cholesterol and triglyceride concentrations in each group are shown in Table 5. At 28 and 49 days, birds in group II had significantly lower serum total cholesterol and triglyceride concentrations than birds in group I (p <0.05).

Carcass yield, abdominal fat and proximate composition

Carcass yield and abdominal fat are shown in Table 6. Group II had significantly higher percentages of carcass yield than group I, and group I had lower percentages of abdominal fat than group II (p <0.05). The proximate compositions are shown in Table 7. In leg and breast cuts, group II had significantly lower percentages of fat than group I (p <0.05).

Jejunum and ileum IgA concentration

Jejunum and ileum IgA concentration for each group is shown in Table 8. At 49 days, birds in group II had significantly greater jejunum and ileum IgA concentration than birds in group I (p <0.05).

Discussion

In the current study, supplemental probiotics (AVITEAM, Toa Pharmaceutical Co. Ltd., Tokyo, Japan) containing B. mesentericus TO-A at 1 x 10⁸ CFU g⁻¹, C. butyricum TO-A at 1 x 10⁸ CFU g⁻¹, and S. faecalis T-110 at 1 x 10⁸ CFU g⁻¹ significantly improved the growth performance and feed conversion ratio in broiler chickens (p <0.05). Many studies have shown that probiotics promote the growth of broiler chickens 19–22 and improve the feed conversion ratio 23–25. The mechanisms through which the probiotics promote growth performance and improve the feed conversion ratio are considered to be complex. One of the reasons for this is that the probiotics maintain gut health, which is conducive to improved growth performance 26. With respect to gut health, it has been reported that probiotics containing B. mesentericus TO-A, C. butyricum TO-A and S. faecalis T-110 are effective for the prevention of the growth of enterohaemorrhagic Escherichia coli, which is harmful to the intestine of infant rabbits 27. Furthermore, it has been reported that this particular probiotic composition produces butyric acid, which promotes the restoration of intestinal mucosal cells 28.

In support of past studies, the current study suggested that probiotics improve the average daily gain and feed conversion ratio by the maintenance of gut health in broiler chickens.

In the current study, group II had a significantly lower serum total cholesterol and triglyceride concentration than group I (p <0.05). Generally, serum total cholesterol and triglyceride concentrations have been an index of lipid metabolism. A similar reduction in serum cholesterol concentration has been found in broilers 29, layers 30, rats 31 and humans 32 fed diets supplemented with Lactobacillus. Reduction in serum triglyceride concentration has been found in rats fed diets supplemented with probiotics containing B. mesentericus, C. butyricum and S. faecalis 33. The decrease in cholesterol concentration could be due to cholesterol assimilation by Lactobacillus cells 34, 35 or due to the co-precipitation of cholesterol with deconjugated bile salts 36. In the current study, it is suggested that by the same mechanism, group II had significantly lower percentages of fat in abdominal, leg and breast cuts than group I.

To the best of our knowledge, there were very few studies reporting on the use of B. mesentericus, C. butyricum and S. faecalis for the prevention of deposition of fat in broiler chickens.

Havenaar and Spanhaak 37 reported that probiotics stimulate the immunity of chickens in two ways: 1) flora from the probiotic migrate throughout the gut wall and multiply to a limited extent or 2) antigen released by dead microorganisms are absorbed; thus, stimulating the immune system. It has been reported that probiotics containing B. mesentericus TO-A, C. butyricum TO-A and S. faecalis T-110 cause an increase in Ig productivity of mesenteric lymph node in rats 33, stimulation of the Th1 immune response in PBMC and dendritic cells 38 and an influx of CD8+ T cells into the intestinal mucosa, which may enhance the intestinal immunity by CD8+ T cells in young chicks 39, but there were no reports that B. mesentericus, C. butyricum and S. faecalis improved gut mucosal immunity in broiler chicken that had mature functions of immunity. In the current study, probiotics included B. mesentericus, C. butyricum and S. faecalis improved jejunum and ileum IgA concentrations significantly at 49 days in broiler chickens. In general, broiler chickens at 49 days had mature functions of immunity. To the best of our knowledge, there were very few studies reporting that the B. mesentericus, C. butyricum and S. faecalis improved gut mucosal immunity in broiler chicken that had mature functions of immunity. The responsible mechanisms remain unclear; however, it has been suggested that probiotics stimulate gut immunity and maintain gut health.

Conclusions

In conclusion, the results of the current study indicated that probiotics containing B. mesentericus TO-A, C. butyricum TO-A and S. faecalis T-110 promote growth performance (average daily gain, feed conversion ratio, carcass yield), gut mucosal immunity (jejunum and ileum IgA concentration) and decrease abdominal and intramuscular fat in broiler chicken. It is therefore suggested that the use of probiotics (AVITEAM) would be advantageous to the poultry industry.

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