Optimization of Emulsification Conditions on Ethanol Extract of Taiwanese Green Propolis Using Polysorbate and Its Immunomodulatory Effects in Broilers.

Felix Shih-Hsiang Hsiao,Yeong-Hsiang Cheng,Chia-Jung Tsai,Yu-Hsiang Yu,Yi-Hsuan Chien,Clara Ajeng Artdita,Yue-Wen Chen,Kuo-Feng Hua

doi:10.3390/ani12040446

Felix Shih-Hsiang Hsiao, Yeong-Hsiang Cheng + Show 6 more

Open Access

https://doi.org/10.3390/ani12040446

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Abstract

Simple SummaryTaiwanese green propolis (TGP) ethanol extract has been shown to have a wide range of biological activities, such as antimicrobial and anti-inflammatory properties. However, extraction using ethanol limits the use of TGP as an ingredient in animal feeds. In addition, the effect of TGP ethanol extract on immunomodulation in broilers is still unclear. In order to increase the utilization of TGP ethanol extract in poultry productivity, this study aimed to establish the optimal emulsification conditions for TGP ethanol extract using polysorbate and investigate its effectiveness in improving immune response in broilers.Beeswax and resin are the main components of propolis, both of which are hydrophobic. The use of emulsifiers helps to improve the extraction of active propolis compounds and makes them more widely used. In this study, we investigated the optimal parameters for the emulsification of Taiwanese green propolis (TGP) using different polysorbates (polysorbate-20, polysorbate-60, and polysorbate-80) and evaluated the effects on the immunomodulatory response in broilers. The results showed that 4 mg/mL of TGP in combination with 2% polysorbate-60 at 60 °C for 60 min significantly decreased the undissolved particle size of ethanol extract of TGP during the emulsification. The bioactive compounds of TGP, the propolins (C, D, F, G, and H), were also detected after emulsification. Supplementation of emulsified TGP (eTGP) in the drinking water of broilers before and after vaccination significantly enhanced the antibody titer response to infectious bronchitis virus at 28 days of age. In the lipopolysaccharide-challenged model, supplementation of eTGP in the drinking water of broilers decreased pro-inflammatory gene expression and increased anti-inflammatory gene expression. These results together suggested that the polysorbate-60 could effectively emulsify the ethanol extract of TGP. Moreover, eTGP could be used as a vaccine adjuvant and an immunomodulator to improve the immune response of broilers.

Highlights

The use of antibiotic growth promoters (AGP) in animal feed has been banned by European nations since 2006, meaning researchers are still looking for AGP alternatives
We first determined the emulsification efficiency between different polysorbates, and 2 mg/mL Taiwanese green propolis (TGP) ethanol extract was used as test material
TGP300 and TGP600 showed no differences in the inhibition of tumor necrosis factor α (TNF-α) and inducible nitric oxide synthase (iNOS) expression levels induced by LPS

Summary

Introduction

The use of antibiotic growth promoters (AGP) in animal feed has been banned by European nations since 2006, meaning researchers are still looking for AGP alternatives. Flavonoids and phenolic esters are the most biologically active main components of propolis [9,10,11]; the contents of these components differ depending on the method of propolis extraction [10,12]. A variety of organic solvents such as ethanol have been used to extract a large number of total flavonoids and phenolic propolis compounds [10,14,15]. It is worth noting that this extraction method limits the use of propolis as an ingredient in animal feeds. Emulsifiers have both hydrophilic and lipophilic substances, which can mix water and oil into a stable emulsion [16]. Only a few studies have explored the effectiveness of polysorbates as emulsifiers for ethanol extracts of propolis

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