Abstract

Salmonella enterica serovar (ser.) Enteritidis (S. Enteritidis) is a foodborne pathogen often associated with contaminated poultry products. This study evaluated the anti-adherence and intracellular clearance capability of oligosaccharides extracted from palm kernel cake (PKC), a by-product of the palm oil industry, and compared its efficacy with commercial prebiotics— fructooligosaccharide (FOS) and mannanoligosaccharide (MOS)—against S. Enteritidis in vitro. Based on the degree of polymerization (DP), PKC oligosaccharides were further divided into ‘Small’ (DP ≤ 6) and ‘Big’ (DP > 6) fractions. Results showed that the Small and Big PKC fractions were able to reduce (p < 0.05) S. Enteritidis adherence to Cancer coli-2 (Caco-2) cells at 0.1 mg/ mL while MOS and FOS showed significant reduction at 1.0 mg/mL and 10.0 mg/mL, respectively. In terms of S. Enteritidis clearance, oligosaccharide-treated macrophages showed better S. Enteritidis clearance over time at 50 µg/mL for Small, Big and MOS, while FOS required a concentration of 500 µg/mL for a similar effect. This data highlights that oligosaccharides from PKC, particularly those of lower DP, were more effective than MOS and FOS at reducing S. Enteritidis adherence and enhancing S. Enteritidis clearance in a cell culture model.

Highlights

  • The human gastrointestinal tract is home to a vast number of microbes, with the large intestine containing up to 1011 bacterial cells per gram of colonic content [1,2]

  • In light of the above knowledge and based on the mode of Salmonella infection starting from the epithelial cells of the intestinal lining to the macrophages of the lamina propria, this study evaluated the protective effects of different sized oligosaccharides extracted from palm kernel cake (PKC) against Salmonella Enteritidis in vitro and compared their efficacy with two commercial prebiotics—FOS and MOS

  • High Performance Liquid Chromatography (HPLC) coupled with an aminopropyl column and a Refractive Index Detector (RID) was used to ensure that the pooled oligosaccharide fractions contained oligosaccharides of different compositions from one another

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Summary

Introduction

The human gastrointestinal tract is home to a vast number of microbes, with the large intestine containing up to 1011 bacterial cells per gram of colonic content [1,2]. Dysbiosis, i.e., the disruption of balance within the gut microbiota due to antibiotics, diets and/or infections may lead to an overgrowth of pathogenic microbes [5,6,7] Salmonella, those of the serovar Enteritidis, is an example of a commonly encountered foodborne pathogen that is often associated with the consumption of contaminated poultry meat and eggs [8]. In light of the above knowledge and based on the mode of Salmonella infection starting from the epithelial cells of the intestinal lining to the macrophages of the lamina propria, this study evaluated the protective effects of different sized oligosaccharides extracted from PKC against Salmonella Enteritidis in vitro and compared their efficacy with two commercial prebiotics—FOS and MOS

Sample Preparation
Size Exclusion Chromatography
Pooling Fractions Based on HPLC Profiles
Molecular Weight Determination
Cells and Bacterial Culture Conditions
Adherence Inhibition Study Using Caco-2 Cells
LDH Analysis of Caco-2 Cells
Intracellular Salmonella Clearance Using U-937
Statistical Analysis
HPLC Profiles of PKC Oligosaccharides
Molecular Weights of Oligosaccharides
Salmonella Enteritidis Adherence Inhibition to Caco-2 Cells
Effects of Oligosaccharides on the Release of LDH in LPS-Induced Caco-2 Cells
Intracellular Clearance of Salmonella Enteritidis in U-937 Macrophages

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