The in vitro faecal evaluation of prebiotic effects of rooibos phenolic compounds on the gut microbiota of vervet monkeys (Chlorocebus pygerythrus)

Abstract

Background: The development of metabolic disease is accompanied by changes in gut microbiota phenotype, including a decrease of beneficial bacteria and increase of pernicious bacteria of the gastrointestinal tract. A Western (high-fat and high-sugar) diet, sedentary lifestyle and altered gut microbiota diversity have been associated with an increased risk of developing metabolic diseases such as type 2 diabetes and its associated risk factor, obesity. Many researchers have studied the link between the gut microbiota and diet. Hence our in vitro study is aimed at investigating the potential prebiotic effect of an aspalathin-rich unfermented rooibos extract, Afriplex GRT™ and aspalathin on the faecal bacterial diversity of vervet monkeys fed Western diet. Methodology: A total of six vervet monkeys (Chlorocebus pygerythrus) were selected from monkeys fed either a maize based normal diet (standard diet group; n=3) or a high fat diet (Western diet group; n=3) for more than 5-years. Faecal samples were collected from the animals in both groups at the Primate Unit and Delft Animal Centre (PUDAC) between 7 – 9 AM. Faecal samples from the two groups were divided into culture-independent baseline samples (before culture) and culture-dependent samples (after anaerobic culture). The culture-dependent samples were cultured under anaerobic conditions at 37°C for 10 hours, with or without Afriplex GRT™ extract or aspalathin. Bacterial genomic DNA (gDNA) was extracted from all samples using the NucleoSpin® DNA Stool extraction kit. Purified gDNA was sent for metagenomic sequencing for 16S rRNA gene analysis of microbial diversity using an Ion Torrent Next-generation Sequencing platform. Results: Results indicated that the Western diet affects the abundance of several bacterial species. Afriplex GRT™ and aspalathin significantly enhanced the relative abundance of health promoting butyrate-producing bacteria such as Faecalibacterium prausnitzii in both standard and Western diet groups (p= 0.02 and p=0.04, respectively). A similar trend was observed in other beneficial bacteria such as Eubacterium spp., Sutterella spp., and Dorea longicatena. Conclusion: Based on the data observed, it can be suggested that Afriplex GRT™ has a beneficial prebiotic effect on gut microbiota diversity and gut health.