In ovo Inoculation of Bacillus subtilis and Raffinose Affects Growth Performance, Cecal Microbiota, Volatile Fatty Acid, Ileal Morphology and Gene Expression, and Sustainability of Broiler Chickens (Gallus gallus)

Affiliations

¹ Department of Animal Production, Faculty of Agriculture, Al-Azhar University, Cairo, Egypt.
² Department of Dairy Science and Food Technology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, India.
³ Department of Animal and Poultry Production, Faculty of Agriculture, Damanhour University, Damanhour, Egypt.
⁴ Sustainable Agriculture Research Group, Department of Agriculture, Faculty of Environmental Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
⁵ Department of Pharmacology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.
⁶ Laboratory of Biotechnology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.
⁷ Biology Department, College of Science, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.
⁸ Avian and Rabbit Diseases Department, Faculty of Veterinary Medicine, Benha University, Banha, Egypt.
⁹ Agricultural Botany Department, Faculty of Agriculture (Saba Basha), Alexandria University, Alexandria, Egypt.
¹⁰ Department of Agricultural Microbiology, Faculty of Agriculture, Zagazig University, Zagazig, Egypt.
¹¹ Biochemistry Department, Faculty of Agriculture, Zagazig University, Zagazig, Egypt.
¹² Biological Applications Department, Nuclear Research Center, Egyptian Atomic Energy Authority, Abu Zaabal, Egypt.

PMID: 35711554
PMCID: PMC9194610
DOI: 10.3389/fnut.2022.903847

In ovo Inoculation of Bacillus subtilis and Raffinose Affects Growth Performance, Cecal Microbiota, Volatile Fatty Acid, Ileal Morphology and Gene Expression, and Sustainability of Broiler Chickens (Gallus gallus)

Abdelrazeq M Shehata et al. Front Nutr. 2022.

. 2022 May 31:9:903847.

doi: 10.3389/fnut.2022.903847. eCollection 2022.

Authors

Affiliations

¹ Department of Animal Production, Faculty of Agriculture, Al-Azhar University, Cairo, Egypt.
² Department of Dairy Science and Food Technology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, India.
³ Department of Animal and Poultry Production, Faculty of Agriculture, Damanhour University, Damanhour, Egypt.
⁴ Sustainable Agriculture Research Group, Department of Agriculture, Faculty of Environmental Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
⁵ Department of Pharmacology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.
⁶ Laboratory of Biotechnology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.
⁷ Biology Department, College of Science, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.
⁸ Avian and Rabbit Diseases Department, Faculty of Veterinary Medicine, Benha University, Banha, Egypt.
⁹ Agricultural Botany Department, Faculty of Agriculture (Saba Basha), Alexandria University, Alexandria, Egypt.
¹⁰ Department of Agricultural Microbiology, Faculty of Agriculture, Zagazig University, Zagazig, Egypt.
¹¹ Biochemistry Department, Faculty of Agriculture, Zagazig University, Zagazig, Egypt.
¹² Biological Applications Department, Nuclear Research Center, Egyptian Atomic Energy Authority, Abu Zaabal, Egypt.

PMID: 35711554
PMCID: PMC9194610
DOI: 10.3389/fnut.2022.903847

Abstract

Banning antibiotic growth promoters has negatively impacted poultry production and sustainability, which led to exploring efficient alternatives such as probiotics, probiotics, and synbiotics. Effect of in ovo injection of Bacillus subtilis, raffinose, and their synbiotics on growth performance, cecal microbial population and volatile fatty acid concentration, ileal histomorphology, and ileal gene expression was investigated in broilers (Gallus gallus) raised for 21 days. On 300 h of incubation, a total of 1,500 embryonated eggs were equally allotted into 10 groups. The first was non-injected (NC) and the remaining in ovo injected with sterile distilled water (PC), B. subtilis 4 × 10⁵ and 4 × 10⁶ CFU (BS1 and BS2), Raffinose 2 and 3 mg (R1 and R2), B. subtilis 4 × 10⁵ CFU + raffinose 2 mg (BS1R1), B. subtilis 4 × 10⁵ CFU + raffinose 3 mg (BS1R2), B. subtilis 4 × 10⁶ CFU + raffinose 2 mg (BS2R1), and B. subtilis 4 × 10⁶ CFU + raffinose 3 mg (BS2R2). At hatch, 60 chicks from each group were randomly chosen, divided into groups of 6 replicates (10 birds/replicate), and fed with a corn-soybean-based diet. In ovo inoculation of B. subtilis and raffinose alone or combinations significantly improved body weight, feed intake, and feed conversion ratio of 21-day-old broilers compared to NC. Cecal concentrations of butyric, pentanoic, propionic, and isobutyric acids were significantly elevated in R1, R2, BS2R1, and BS2R2, whereas isovaleric and acetic acids were significantly increased in R1 and BS2R1 compared to NC. Cecal microbial population was significantly altered in treated groups. Ileal villus height was increased (p < 0.001) in BS1, R2, and BS2R2 compared to NC. The mRNA expression of mucin-2 was upregulated (p < 0.05) in synbiotic groups except for BS1R1. Vascular endothelial growth factor (VEGF) expression was increased (p < 0.05) in BS2, R1, BS1R1, and BS1R2 compared to NC. SGLT-1 expression was upregulated (p < 0.05) in all treated birds except those of R1 group compared to NC. The mRNA expressions of interleukin (IL)-2 and toll-like receptor (TLR)-4 were downregulated (p < 0.05) in BS2 and R1 for IL-2 and BS1R1 and BS2R2 for TLR-4. It was concluded that in ovo B. subtilis, raffinose, and synbiotics positively affected growth performance, cecal microbiota, gut health, immune responses, and thus the sustainability of production in 21-day-old broilers.

Keywords: bioactive compounds; broiler chickens; gene expression; gut microbiota; ileal architecture; in ovo feeding; sustainability; volatile fatty acid.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

**FIGURE 1**
Effect of *in ovo* inclusion of *Bacillus subtilis*, raffinose, and their synbiotics on ileal histomorphometry of 21-day-old broilers. Each number on the figure means the following groups: 1 = non-injected group; 2 = sterile distilled water; 3 = *Bacillus subtilis* 4 × 10⁵/egg; 4 = *Bacillus subtilis* 4 × 10⁶/egg; 5 = Raffinose 2 mg/egg; 6 = Raffinose 3 mg/egg; 7 = (*Bacillus subtilis* 4 × 10⁵ + Raffinose 2 mg)/egg; 8 = (*Bacillus subtilis* 4 × 10⁵ + Raffinose 3 mg)/egg; 9 = (*Bacillus subtilis* 4 × 10⁶ + Raffinose 2 mg)/egg; 10 = (*Bacillus subtilis* 4 × 10⁶ + Raffinose 3 mg)/egg. Images were captured with light microscopy. Scale bar indicates 500 μm.

**FIGURE 2**
Effect of *in ovo* inclusion of *Bacillus subtilis*, raffinose, and their synbiotics on ileal gene expression of 21-day-old broilers. **(A,B)** Intestinal function-related, **(C,D)** nutrient transporter genes, and **(E,F)** immune-related genes. NC, non-injected group; PC, sterile distilled water; BS1 = *Bacillus subtilis* 4 × 10⁵/egg; BS2 = *Bacillus subtilis* 4 × 10⁶/egg; R1 = Raffinose 2 mg/egg; R2 = Raffinose 3 mg/egg; BS1R1 = (*Bacillus subtilis* 4 × 10⁵ + Raffinose 2 mg)/egg; BS1R2 = (*Bacillus subtilis* 4 × 10⁵ + Raffinose 3 mg)/egg; BS2R1 = (*Bacillus subtilis* 4 × 10⁶ + Raffinose 2 mg)/egg; BS2R2 = (*Bacillus subtilis* 4 × 10⁶ + Raffinose 3 mg)/egg. Data are presented as the mean values with their standard errors. Gene expression differences were evaluated using fold changes compared to the non-injected group. The significance was declared at p < 0.05. Bars with different letters represent significant differences among the different groups.

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In ovo Inoculation of Bacillus subtilis and Raffinose Affects Growth Performance, Cecal Microbiota, Volatile Fatty Acid, Ileal Morphology and Gene Expression, and Sustainability of Broiler Chickens (Gallus gallus)

Affiliations

In ovo Inoculation of Bacillus subtilis and Raffinose Affects Growth Performance, Cecal Microbiota, Volatile Fatty Acid, Ileal Morphology and Gene Expression, and Sustainability of Broiler Chickens (Gallus gallus)

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

LinkOut - more resources

Full Text Sources