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. 2024 Nov;103(11):104237.
doi: 10.1016/j.psj.2024.104237. Epub 2024 Aug 22.

Dietary supplementation of microencapsulated botanicals and organic acids enhances the expression and function of intestine epithelial digestive enzymes and nutrient transporters in broiler chickens

Andrea Toschi  1 , Liang-En Yu  2 , Sofia Bialkowski  2 , Lydia Schlitzkus  2 , Ester Grilli  3 , Yihang Li  4
Affiliations

Dietary supplementation of microencapsulated botanicals and organic acids enhances the expression and function of intestine epithelial digestive enzymes and nutrient transporters in broiler chickens

Andrea Toschi et al. Poult Sci. 2024 Nov.

Abstract

Organic acids and botanicals have shown protective effects on gut barrier and against inflammation in broilers. However, their effects on intestinal digestive enzymes and nutrients transporters expression and functions have not been fully studied. The objective of this study was to understand how a microencapsulated blend of botanicals and organic acids affected intestinal enzyme activities and nutrient transporters expression and functions in broilers. A total of 288 birds were assigned to a commercial control diet or diet supplemented with 500 g/MT (metric ton) of the microencapsulated additive. Growth performance was recorded weekly. At d 21 and d 42, jejunum and ileum were isolated for enzyme (maltase, sucrase, and aminopeptidase) and transporter (SGLT1, GLUT2, GLUT1, EAAT3, B0AT1, and PepT1) analyses. Jejunum specific nutrients (glucose, alanine, and glutamate) transport activities were evaluated by Ussing chamber. Protein expression of nutrient transporters in small intestine were measured in mucosa and brush-border membrane (BBM) samples by western blot. Intestinal gene expression of the transporters was determined by RT-PCR. Statistical analysis was performed using Student's t-test comparing the supplemented diet to the control. The feed efficiency was significantly improved through the study period in the supplemented group (P ≤ 0.05). Significant changes of intestinal histology were shown in both jejunum (P ≤ 0.10) and ileum (P ≤ 0.05) after 21 d of treatment. At d21, jejunal maltase activity was upregulated (P ≤ 0.10). The Ussing chamber transport of glucose and alanine was increased, which was in line with increased gene expression (GLUT2, GLUT1, EAAT3, and B0AT1) (P ≤ 0.10 and P ≤ 0.05, respectively) and BBMV protein levels (B0AT1, P < 0.10). At d21, ileal sucrase and maltase activities were upregulated (P ≤ 0.05). Increased expressions of GLUT1, EAAT3, and B0AT1 were observed in both mRNA and protein levels (P ≤ 0.05). Similar pattern of changes was also shown at d42 of age. Our results suggest that feeding microencapsulated additives improves intestinal nutrient digestion and transporter expression and function in broilers, thereby enhancing feed efficiency.

Keywords: botanical; broiler; digestive enzyme; intestinal nutrient transport; organic acid.

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Conflict of interest statement

DISCLOSURES The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ester Grilli reports financial support was provided by Vetagro S.p.A. Ester Grilli reports a relationship with Vetagro Inc. that includes: board membership. Ester Grilli reports a relationship with University of Bologna that includes: employment. Andrea Toschi reports a relationship with Vetagro S.p.A. that includes: employment.

Figures

Figure 1
Figure 1
Growth performance results of birds fed with control or supplemented diet. ADG = Average daily gain; ADFI = average daily feed intake; FCR = feed conversion ratio. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 2
Figure 2
Digestive enzyme activity in jejunum of birds fed with control or supplemented diet. Maltase activity in BBMV and mucosa and sucrase activity in BBMV and mucosa are reported. BBMV = brush-border membrane vesicles. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 3
Figure 3
Nutrient transporter function in jejunum of birds fed with control or supplemented diet. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #Indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 4
Figure 4
Nutrient transporter protein levels in jejunum of birds fed with control or supplemented diet. BBMV = brush-border membrane vesicles; SGLT1 = sodium/glucose cotransporter 1; B0AT1 = Sodium-dependent neutral amino acid transporter; EAAT3 = excitatory AAs transporter 3. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 5
Figure 5
Nutrient transporter mRNA levels in jejunum of birds fed with control or supplemented diet. SGLT1 = sodium/glucose cotransporter 1; GLUT1 = glucose transporter 1; GLUT 2 = glucose transporter 2; EAAT3 = excitatory AAs transporter 3; B0AT1 = Sodium-dependent neutral amino acid transporter; PepT1 = peptide transporter 1. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 6
Figure 6
Digestive enzyme activity in ileum of birds fed with control or supplemented diet. Maltase activity in BBMV and mucosa and sucrase activity in BBMV and mucosa are reported. BBMV = brush-border membrane vesicles. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 7
Figure 7
Nutrient transporter protein levels in ileum of birds fed with control or supplemented diet. BBMV = brush-border membrane vesicles; SGLT1 = sodium/glucose cotransporter 1; B0AT1 = Sodium-dependent neutral amino acid transporter; EAAT3 = excitatory AAs transporter 3. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 8
Figure 8
Nutrient transporter mRNA levels in ileum of birds fed with control or supplemented diet. SGLT1 = sodium/glucose cotransporter 1; GLUT1 = glucose transporter 1; GLUT 2 = glucose transporter 2; EAAT3 = excitatory AAs transporter 3; B0AT1 = Sodium-dependent neutral amino acid transporter; PepT1 = peptide transporter 1. Difference between 2 groups at each growth period was labeled. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
Figure 9
Figure 9
Serum glucose level in birds fed with control or supplemented diet. *Indicates significant difference (P ≤ 0.05), #indicates tendency (P ≤ 0.10). Dietary treatments were as follows: CTR, basal diet; OA+B, basal diet + 500 ppm microencapsulated organic acids and botanicals.
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