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. 2020 Aug 13;15(8):e0237505.
doi: 10.1371/journal.pone.0237505. eCollection 2020.

Excreta biomarkers in response to different gut barrier dysfunction models and probiotic supplementation in broiler chickens

Reza Barekatain  1   2 , Gordon S Howarth  2 , Nicky-Lee Willson  2 , David Cadogan  3 , Stuart Wilkinson  3
Affiliations

Excreta biomarkers in response to different gut barrier dysfunction models and probiotic supplementation in broiler chickens

Reza Barekatain et al. PLoS One. .

Abstract

Increased intestinal permeability (IP) and inflammation are both linked with functionality of the intestinal barrier and in particular enterocytes. Currently, almost all assessment methods of the intestinal barrier function are invasive. The present study aimed to quantify selected proteins as novel biomarkers in excreta of broiler chickens to facilitate non-invasive assessment of gut barrier function using enzyme-linked immunosorbent assays (ELISA). It was further hypothesised that probiotics as feed additives may counteract gut barrier dysfunction. A 3 × 2 factorial arrangement of treatments was used with the main factors being gut barrier dysfunction models (control, rye-based diet, and dexamethasone-DEX) with and without probiotic supplementation (a three-strain Bacillus) using 72 male Ross 308 day-old chickens. Each of the 6 experimental treatments was replicated 12 times. On d 21 of age, fluorescein isothiocyanate dextran (FITC-d) uptake into serum was examined to test IP. Fresh excreta samples were collected on d 20. The biomarkers included alpha-1 antitrypsin (A1AT), intestinal fatty acid binding protein (I-FABP), lipocalin-2 (LCN2), fibronectin (FN), intestinal alkaline phosphatase (IAP), ovotransferrin (OVT) and superoxide dismutase [Cu-Zn] (SOD1). Only DEX increased (P<0.001) FITC-d passage to the blood on d 21 of age, indicating a greater IP. The excreta concentrations of A1AT, I-FABP and SOD1 were unaltered by the experimental treatments. DEX increased (P<0.05) FN concentration in excreta compared with control birds. Conversely, inclusion of rye in the diet reduced (P<0.05) FN but increased (P<0.001) OVT in excreta. Independently, DEX decreased IAP (P<0.05) in excreta compared with control and rye-fed birds. The excreta concentration of LCN2 tended (P = 0.086) to increase in birds injected by DEX. There was no demonstrable effect of probiotic addition on any of the studied parameters. Among the tested biomarkers, FN, IAP, and LCN2 revealed promise as biomarkers of intestinal barrier function quantified by ELISA kits.

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Conflict of interest statement

D.C. is a member of the Industry Committee of PoultryHub Australia. D.C. and S.W. are both employed by the Feedworks Ltd Pty. The specific roles of these authors are articulated in the ‘author contributions’ section. Feedworks did not provide funding for the study and this commercial affiliation does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Serum FITC-d concentration of broilers subjected to two leaky gut models with and without probiotic.
Error bars represent standard error of the mean (SEM).
Fig 2
Fig 2. Concentration of Alpha 1 antitrypsin in excreta samples of broilers (n = 72) for the main effects of gut barrier dysfunction models (P>0.05) and probiotic supplementation (P>0.05).
Error bars represent standard error of the mean (SEM).
Fig 3
Fig 3. Concentration of fibronectin in excreta samples (n = 72) for the main effects of gut barrier dysfunction models (P<0.05) and probiotic supplementation (P>0.05).
The error bars represent standard error of the mean (SEM). Bars with differing superscripts are statistically different (P<0.05).
Fig 4
Fig 4. Concentration of intestinal fatty acid binding protein in excreta samples (n = 72) for the main effects of gut barrier dysfunction models (P>0.05) and probiotic supplementation (P>0.05).
Error bars represent standard error of the mean (SEM).
Fig 5
Fig 5. Concentration of Lipocalin 2 in excreta samples (n = 72) for the main effects of gut barrier dysfunction models and probiotic supplementation.
Error bars represent standard error of the mean (SEM).
Fig 6
Fig 6. Concentration of ovotransferrin in excreta samples (n = 72) for the main effects of gut barrier dysfunction models and probiotic supplementation.
Error bars represent standard error of the mean (SEM).
Fig 7
Fig 7. Concentration of intestinal alkaline phosphatase in excreta samples (n = 72) for the main effects of gut barrier dysfunction models and probiotic supplementation.
Error bars represent standard error of the mean (SEM). Bars with differing superscripts are statistically different (P<0.05).
Fig 8
Fig 8. Concentration of superoxide dismutase [Cu-Zn] in excreta samples (n = 36) for the main effects of gut barrier dysfunction models (P>0.05) and probiotic supplementation (P>0.05).
Error bars represent standard error of the mean (SEM).
See this image and copyright information in PMC

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