Abstract

Previously, we demonstrated that rectal swabs provide a legitimate alternative to faecal sampling for the assessment of the intestinal microbiota in young piglets. However, we also reported that mucosa-adhered microbial populations were more represented in rectal swabs compared to faecal samples, albeit to a degree that varied per swab-sample. Here, we explored the possibility to exploit this variable enrichment of adhered populations in the rectal swabs to assess the impact of diet on mucosa-adhered microbiota in pre-weaning piglets. Paired samples of rectal swabs and colon luminal contents were collected from piglets just before weaning during two independent but similarly designed animal experiments [n = 28 piglets (experiment 1); n = 16 piglets (experiment 2)], with an early feeding treatment (EF) group that had access to customised fibrous feed in addition to sow’s milk and a control (CON) group exclusively reared on sow’s milk. The intestinal microbiome composition in rectal swabs and colon samples collected at 29 days of age were subjected to metataxonomic analysis. The results identified the genera Escherichia-Shigella, Anaerococcus, Peptostreptococcus, Enterococcus, Trueperella, Actinomyces, and Peptoniphilus as discriminative taxa enriched in rectal swabs compared to colon. Apart from Escherichia-Shigella (10–11% average relative abundance), most of these mucosa-adhered microbial genera display relatively low abundance. Rectal swab microbiota was found to be more variable, which is likely due to variable enrichment of mucosa-adhered microbes. Although almost exclusively driven by one of the experiments, the post-weaning diarrhoea-associated taxa Escherichia-Shigella, was enriched in CON compared to the EF group, suggesting that early life feeding may suppress post-weaning-diarrhoea-related problems in piglets. Our findings demonstrate that rectal swabs allow the investigation of the mucosa-adhered microbial populations as a function of dietary treatment in piglets. This offers opportunities to further study dietary approaches that suppress the abundance of the post-weaning diarrhoea associated adherent microbes like Escherichia-Shigella. Furthermore, we demonstrate that the paired swab-colon microbiota information (obtained from a subset of animals) can predict the mucosa-adhered populations or “mucosity factor” in rectal swab samples, facilitating the analysis of the adhered microbiota in large animal cohort studies using readily obtainable rectal swabs.

Highlights

  • Choice of a suitable sample type is important for reliable microbiota analysis

  • We have shown that rectal swab is a legitimate alternative to faeces, to study the early life microbiome development in piglets, we recognised that the mucosa-adhered microbial groups were more represented in rectal swabs to a variable degree (Choudhury et al, 2019)

  • The present study aims to bring our previous findings one step further, by evaluating the impact of pre-weaning fibrous diet (Choudhury et al, 2021a) on the mucosa-adhered microbial population using paired colon and rectal swab samples obtained from the same piglets at the end of lactation

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Summary

Introduction

Choice of a suitable sample type is important for reliable microbiota analysis. This is because microbiome composition can be influenced by the chosen sample type, due to high niche-specificity of residing microbial communities. Multiple niches or micro-environments exist in the intestinal tract, such as the intestinal lumen, epithelial mucus layer, or the mucosal tissues, which encompass differential abundance of microbial populations (Jones et al, 2018) This notion is supported by several studies that reported differences in the microbial populations found to be mucosa-associated or in the lumen of the gut in various mammals, including humans (Carroll et al, 2010; Burrough et al, 2017; Jones et al, 2018; Zhang et al, 2018; Mottawea et al, 2019; Wu et al, 2020; Klymiuk et al, 2021). We implied that contrary to faecal samples, rectal swabs have the potential to reflect the mucosa-adhered population albeit to an unpredictable and quite variable degree per sample (Choudhury et al, 2019)

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