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current biomarkers require invasive sampling and specific laboratory analyses (ELISA, RT-qPCR, HPLC, etc.) limiting their use in the research field. Nevertheless, nowadays, different reports about the development of rapid and non-invasive techniques that could be applied directly on the farm have been published; with the most promising ones based on the detection of molecules ending up in the excreta and litter. In summary, the analysis of several markers is mandatory when evaluating the efficacy of a feed additive focused on gut health, in order to deeply understand its mode of action, which is usually quite complex.


Use of olive bioactives in poultry Several studies have evaluated the effects of standardised mixtures of olive bioactives in poultry, with special emphasis on gut health. Animals who received olive bioactives in their feeds showed positive results on performance versus the control group (Figure 2), especially under challenging situations. Therefore, to better understand the mode of action of this family of products, several biomarkers (in vitro and in vivo) have been measured to have a wider vision of their biological effects into the animal, with especial focus on gut function and the inflammatory status of the animal. Preliminary cell studies demonstrated the potential activity of these olive bioactives on reducing oxidative stress (augmented catalase and glutathione gene expressionin IPEC-J2 cells), reducing inflammation (prevented IL-1β and iNOS gene overexpression in Raw 264.7 cells) and improving gut integrity (improved TEER and upregulated tight junctions gene expression in IPEC-J2 cells). In these trials, cells were immune challenged with LPS and data showed prevention of negative consequences derived from the LPS challenge after pre-incubation with olive bioactives. To explore the mode of action of a specific formulation based on olive bioactives in broiler chicken diets and their potential benefits, Lucta and its collaborators performed different in vivo trials. In a first trial, birds supplemented with olive bioactives improved animal growth as a result of its immunomodulatory effect at the gut level. Additionally, these results were not different from those obtained with the positive control with monensin (Figure 3). The same olive bioactives formulation was evaluated again in an in vivo intestinal inflammatory model based on feed restriction. After a short feed restriction period, broiler chickens displayed a loss of intestinal integrity, as evidenced by a decrease in the gene expression of Claudin-1, a lower villus length/crypt depth ratio, and a higher serum lactulose/mannitol ratio. In addition, the challenge increased intestinal inflammation as suggested by the upregulation of the gene expression of pro-inflammatory cytokine IL-8 and the Toll-Like Receptor 4 in the ileum. The supplementation of olive bioactives in the diet was able to prevent some of these negative effects, resulting in a


  


Average Daily Gain 120   


110 100 90 80


70 60


 


 Days of age 


 





   2.5


MON OB 2 * 1.5 1 0.5 * 0 IL-8 *P < 0.05 compared to control.


TFG-beta4 Biomarker


BU-1 * * *


better intestinal functionality (higher serum mannitol concentration and lower duodenal crypth depth), a modulation of the immune system (higher gene expression of the B-cell marker Bu-1) and an attenuated local inflammation (decreased gene expression of IL-8).


Promising tool The use of specific formulations of olive bioactive compounds in broiler chicken diets may be considered as a promising tool to deal with challenging situations associated to our poultry production systems thanks to boost antioxidant, anti-inflammatory and immunomodulatory activities in broilers to stabilise their physiological condition and support animal welfare.


▶ GUT HEALTH | DECEMBER 2020 83


Fold change


g/d


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