Supplementation of protected sodium butyrate alone or in combination with essential oils modulated the cecal microbiota of broiler chickens challenged with coccidia and Clostridium perfringens.

Authors: BORTOLUZZI, CRISTIANO - University Of Georgia; Rothrock, Michael; VIEIRA, B - University Of Georgia; MALLO, J - Norel Animal Nutrition; PUYALTO, M - Norel Animal Nutrition; HOFACRE, CHARLES - Southern Poultry Research, Inc; APPLEGATE, TODD - University Of Georgia

Submitted to: Frontiers in Sustainable Food Systems
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/17/2018
Publication Date: 11/6/2019
Citation: Bortoluzzi, C., Rothrock Jr, M.J., Vieira, B.S., Mallo, J.J., Puyalto, M., Hofacre, C., Applegate, T.J. 2019. Supplementation of protected sodium butyrate alone or in combination with essential oils modulated the cecal microbiota of broiler chickens challenged with coccidia and Clostridium perfringens.. Frontiers in Sustainable Food Systems. 2:72. https://doi.org/10.3389/fsufs.2018.00072.
DOI: https://doi.org/10.3389/fsufs.2018.00072

Interpretive Summary: The objective of this study was to determine the effects of protected sodium butyrate (SB), and protected sodium butyrate plus essential oils (carvacrol and ginger; SBEO) on the establishment of the cecal microbiota of broilers induced to necrotic enteritis (NE). Birds were assigned to 4 treatments (8 replicates of 58 birds): (1) unchallenged control; (2) challenged control; (3) challenged and supplemented with SB; (4) challenged and supplemented with SBEO. On d 13, challenged birds were orally inoculated with ~5,000 Eimeria maxima oocysts. On d 18-19, the same birds were exposed to Clostridium perfringens via drinking water (~1x108 cfu). Cecal excreta was collected at d 12, 18, 21, and 28 for microbiota analysis through 16s rRNA sequencing using Illumina platform. All sequence data processing was performed using QIIME v. 1.9.1 software. NE challenge impaired FCR in the overall experimental period (d 1-41) by 5%, but SB and SBEO supplementation completely reverted this effect (p = 0.01). The establishment of the cecal microbiota over time within each experimental group was evaluated. The inclusion of SB alone or in combination with EO contributed significantly to the establishment of the cecal microbiota, as shown by the diversity indexes. The community structure and abundance of the cecal microbiota were significantly different across ages, especially in the groups supplemented with SB and SBEO. As shown in the PCoA analysis, the supplementation of SB led a more stable microbial community and lower between sample variability over time. The cecal microbiota in the four ages evaluated was dominated by bacteria belonging to the phylum Firmicutes, followed by Bacteroidetes and Proteobacteria. The most abundant genera observed in the cecal microbiota were: Ruminococcus, Lactobacillus, and Bacteroides, followed by unclassified Ruminococcaceae and Clostridiales. In the unchallenged control birds, it was observed that Ruminococcus decreased (p = 0.006), whereas Bacteroides and Clostridiales increased (p = 0.02) as the birds aged. In the challenged control group, however, it was also observed that the frequency of Coprococcus and Blautia decreased as birds aged (p = 0.01) and, Clostridiales did not increase as a consequence of in this group of birds. Supplementation of SB, but not SBEO, increased the frequency of Lactobacillus (p = 0.01) on d 12 compared to d 18 and 28, and prevented the reduction in the frequency of Blautia as the birds aged. Nevertheless, supplementation of SB and SBEO contributed to many changes in the predicted functions of the cecal microbiota over time, which was not observed in the unsupplemented birds. SB and SBEO improved growth performance of NE challenged broilers, which may be partially attributed to their modulating effects on the cecal microbiota.

Technical Abstract: The objective of this study was to determine the effects of protected sodium butyrate (SB), and protected sodium butyrate plus essential oils (carvacrol and ginger; SBEO) on the establishment of the cecal microbiota of broilers induced to necrotic enteritis (NE). Birds were assigned to 4 treatments (8 replicates of 58 birds): (1) unchallenged control; (2) challenged control; (3) challenged and supplemented with SB; (4) challenged and supplemented with SBEO. On d 13, challenged birds were orally inoculated with ~5,000 Eimeria maxima oocysts. On d 18-19, the same birds were exposed to Clostridium perfringens via drinking water (~1x108 cfu). Cecal excreta was collected at d 12, 18, 21, and 28 for microbiota analysis through 16s rRNA sequencing using Illumina platform. All sequence data processing was performed using QIIME v. 1.9.1 software. NE challenge impaired FCR in the overall experimental period (d 1-41) by 5%, but SB and SBEO supplementation completely reverted this effect (p = 0.01). The establishment of the cecal microbiota over time within each experimental group was evaluated. The inclusion of SB alone or in combination with EO contributed significantly to the establishment of the cecal microbiota, as shown by the diversity indexes. The community structure and abundance of the cecal microbiota were significantly different across ages, especially in the groups supplemented with SB and SBEO. As shown in the PCoA analysis, the supplementation of SB led a more stable microbial community and lower between sample variability over time. The cecal microbiota in the four ages evaluated was dominated by bacteria belonging to the phylum Firmicutes, followed by Bacteroidetes and Proteobacteria. The most abundant genera observed in the cecal microbiota were: Ruminococcus, Lactobacillus, and Bacteroides, followed by unclassified Ruminococcaceae and Clostridiales. In the unchallenged control birds, it was observed that Ruminococcus decreased (p = 0.006), whereas Bacteroides and Clostridiales increased (p = 0.02) as the birds aged. In the challenged control group, however, it was also observed that the frequency of Coprococcus and Blautia decreased as birds aged (p = 0.01) and, Clostridiales did not increase as a consequence of in this group of birds. Supplementation of SB, but not SBEO, increased the frequency of Lactobacillus (p = 0.01) on d 12 compared to d 18 and 28, and prevented the reduction in the frequency of Blautia as the birds aged. Nevertheless, supplementation of SB and SBEO contributed to many changes in the predicted functions of the cecal microbiota over time, which was not observed in the unsupplemented birds. SB and SBEO improved growth performance of NE challenged broilers, which may be partially attributed to their modulating effects on the cecal microbiota.