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. 2019 Jul 26;7(8):218.
doi: 10.3390/microorganisms7080218.

Developing Gut Microbiota Exerts Colonisation Resistance to Clostridium ( syn. Clostridioides) difficile in Piglets

Łukasz Grześkowiak  1 , Temesgen Hailemariam Dadi  2 , Jürgen Zentek  2 , Wilfried Vahjen  2
Affiliations
Free PMC article

Developing Gut Microbiota Exerts Colonisation Resistance to Clostridium ( syn. Clostridioides) difficile in Piglets

Łukasz Grześkowiak et al. Microorganisms. .
Free PMC article

Abstract

Clostridium (syn. Clostridioides) difficile is considered a pioneer colonizer and may cause gut infection in neonatal piglets. The aim of this study was to explore the microbiota-C. difficile associations in pigs. We used the DNA from the faeces of four sows collected during the periparturient period and from two to three of their piglets (collected weekly until nine weeks of age) for the determination of bacterial community composition (sequencing) and C. difficile concentration (qPCR). Furthermore, C. difficile-negative faeces were enriched in a growth medium, followed by qPCR to verify the presence of this bacterium. Clostridium-sensu-stricto-1 and Lactobacillus spp. predominated the gut microbiota of the sows and their offspring. C. difficile was detected at least once in the faeces of all sows during the entire sampling period, albeit at low concentrations. Suckling piglets harboured C. difficile in high concentrations (up to log10 9.29 copy number/g faeces), which gradually decreased as the piglets aged. Enrichment revealed the presence of C. difficile in previously C. difficile-negative sow and offspring faeces. In suckling piglets, the C. difficile level was negatively correlated with carbohydrate-fermenting bacteria, and it was positively associated with potential pathogens. Shannon and richness diversity indices were negatively associated with the C. difficile counts in suckling piglets. This study showed that gut microbiota seems to set conditions for colonisation resistance against C. difficile in the offspring. However, this conclusion requires further research to include host-specific factors.

Keywords: Clostridioides; Clostridium difficile; enrichment; gut microbiota; pig; qPCR; sequencing; sow; suckling; weaned.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(a) Stacked bar plots showing the average percentage of bacterial taxa in sow and offspring faeces over time, (b) and a Venn diagram depicting the number of bacterial taxa that were unique and shared between sows, suckling, and weaned piglets, as analysed by the 16S rDNA sequencing.
Figure 2
Figure 2
Diversity shown as (a) richness, (b) evenness, and (c) Shannon indices, and (d) a principal component analysis performed using the relative abundance of faecal microbial taxa in sows (green box plots), suckling (violet box plots) and weaned piglets (orange box plots) over time, as analysed by the 16S rDNA sequencing. Dots in figures (a–c) indicate outliers. ap: antepartum; pp: postpartum.
Figure 3
Figure 3
Concentration (mean ± SD, log10 copy/g) of C. difficile in faecal samples from the offspring (n = 8–10) sampled weekly from one until nine weeks of age and analysed using qPCR, without previous sample enrichment. SP: suckling piglets; WP: weaned piglets.
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