Effect of two seaweed polysaccharides on intestinal microbiota in mice evaluated by illumina PE250 sequencing

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Abstract

Effect of polysaccharides from two seaweeds, Porphyra haitanensis and Ulva prolifera, on intestinal microbiota in mice was evaluated by illumina PE250 sequencing. Analysis showed significant structural changes in fecal microbiota among the three sample groups. There were significant differences in the composition of fecal microbiota among the three groups at phylum and genus levels. At the phylum level, the most predominant phylum was Bacteroidetes contributing 58.76%, 73.39%, 75.38% and 64.40% of the fecal microbiota in K, Z, H and D groups respectively, followed by Firmicutes, contributing 37.61%, 23.99%, 21.87% and 30.82% respectively. Many genera were significantly higher in the Z and H group than in the K group, including Prevotellaceae UCG-001 (p < 0.05) and Rikenellaceae RC9 (p < 0.01). In conclusion, our results suggest that polysaccharide type and glycoside may contribute to shaping mice gut microbiota.

Introduction

Intestinal microbiota is the complex community of microorganisms that live in the digestive tracts of humans and other animals [1]. Factors that disrupt the intestinal microbiota include antibiotics, food, stress, and parasites [2,3]. Studies analyses have identified the different bacterial genera in gut microbiota and their associations with nutrient intake [4]. There is an association between the concentration of each microbial community and diet. For example, Prevotella, a genus of Gram-negative bacteria in gut, is related to carbohydrates and simple sugars [5]. Specialist microbes survive on their host's carbohydrate excretions. The composition and diversity of them would change when altering the diet [6]. It is meaningful to study the relation between the diet and intestinal microbiota.

As previously reported, the intake of oligosaccharides or polysaccharide recovered and improve the ecosystem by changing the microbial composition [[7], [8], [9], [10]]. Porphyra haitanensis and Ulva prolifera are two common seaweed. Studies suggested that the polysaccharide from these two seaweeds have excellent physiological activities, including antitumor, immunomodulating, antioxidant and antihyperlipidemic [11,12]. However, scare publication information about their effect on intestinal microbiota. Among a number of approaches for the normalization of RNA sequencing data emerged in the literature, the Illumina sequencing platforms generate up to 100 gigabases of high quality sequence data per lane (HiSeq 4000) or up to 15Gb (MiSeq) [13,14]. The Illumina instruments provide currently the highest yields as well as the highest quality data. Hence, in the present study, two degraded polysaccharides from P. haitanensis and U. prolifera were studied and its effect on intestinal microbiota in mice evaluated by illumina PE250 sequencing were investigated.

Section snippets

Sample preparation and chemicals

P. haitanensis and U. prolifera were separately collected on the coast of Wenzhou and Qingdao. Natural polysaccharides were extracted in hot-water soluble as described previously [15]. The degraded polysaccharide was prepared with the method of H2O2/Vc and the chemical has reported in previous study [16,17]. Two polysaccharides were named after sample Z (P. haitanensis) and H (U. prolifera) in present study.

Animals and treatment

Twenty-four healthy male C57BL/6 mice (8 weeks, 20 ± 2 g; Certificate number, 2015000529984)

Statistics of trimed sequences

Downstream analyses of quality and chimera filtered reads for all the samples were performed using the Usearch software. Paired-end reads was assigned to samples based on their unique barcode and truncated by cutting off the barcode and primer sequence. All the trimed sequences length was shown in Fig. 1. From the figure, the proportion of the high-quality sequence was >95%, which meets the requirements of subsequent sequencing. At the same time, most of the sequence length was between 351 bp

Acknowledgements

This work was supported by funds from Natural Science Foundation of China (No. 31700307), Natural Science Foundation of Zhejiang Province (No. LQ16D060005), the Key Research Program of Frontier Sciences, Chinese Academy of Sciences (No. QYZDB-SSW-DQC023), the Youth Innovation Promotion Association, Chinese Academy of Sciences (No. 2015164), and the Foundation for Huiquan Young Scholar of Institute of Oceanology, Chinese Academy of Sciences (No. 2015).

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