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The impact of probiotics and lactoferrin supplementation on piglet gastrointestinal microbial communities

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Probiotics and lactoferrin are currently being used in neonatal intensive care units in the hopes of reducing rates of sepsis and necrotizing enterocolitis (NEC). While studies have shown that these measures can be clinically beneficial to premature babies, and there are ongoing trials to measure their impact on NEC and sepsis rates, little is known about how they may impact microbiota development. We thus employed a newborn piglet model to assess the impact of feeding probiotics or a combination of probiotics and lactoferrin on development of the gastrointestinal microbiota. Healthy full-term piglets were fed either probiotics alone or probiotics and a bovine lactoferrin supplement over the first weeks of life, and their microbiota profiles were compared with unsupplemented controls. We found that both probiotic and probiotic plus lactoferrin treatments impacted the microbial composition within the gastrointestinal tract, with differing impacts on various regions within the gut. In addition, the impact of probiotics was often reversed by the presence of lactoferrin and both feeding interventions altered the microbiota’s genetic propensity to use ferric versus ferrous ions. These results suggest that iron availability may be a key factor to consider when designing feeding interventions that target the microbiome.

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This work was funded by the Government of Canada through Genome Canada and the Ontario Genomics Institute (Grant No. OGI-067 and OGI-149), CIHR grant number GPH-129340, CIHR grant number MOP-114872, CIHR grant number ECD-144627 and the Ontario Ministry of Economic Development and Innovation (REG1-4450). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Correspondence to Alain Stintzi.

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Supplementary material 1—Principal co-ordinate analysis (PCoA) of piglet microbiota profiles (a–c) and sequencing replicates (d–f), along with median distances between sequencing replicates (g–i) using the Bray Curtis dissimilarity (a, d, g), weighted Unifrac (b, e, h) and unweighted Unifrac (c, f, i). The PCoA plots are colored by piglet batch and the sequencing groups within the replicates are denoted by shapes (PDF 449 kb)


Supplementary material 2—Principal co-ordinate analyses of the piglet intestine microbiota profiles (a–c) and stool microbiota profiles (e–f) using the Bray Curtis dissimilarity (a, d), weighted Unifrac (b, e) and unweighted Unifrac (c, f) colored by either intestine grouping (a–c) or sample date (d–f) with shapes denoting the piglet batch (PDF 257 kb)


Supplementary material 3—Linear regressions with 95% confidence intervals of species richness (a) and evenness (b) in the stool samples over time with the start of the feeding intervention highlighted with a dotted line (PDF 110 kb)

Supplementary material 4—Species richness (a) and evenness (b) across the gastrointestinal tract (PDF 205 kb)


Supplementary material 5—Taxa and KEGG terms identified as being differentially abundant between the three feeding interventions (XLSX 334 kb)

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Grzywacz, K., Butcher, J., Romain, G. et al. The impact of probiotics and lactoferrin supplementation on piglet gastrointestinal microbial communities. Biometals 32, 533–543 (2019).

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