Fecal bacteria and metabolite responses to dietary lysozyme in a sow model from late gestation until lactation

Abstract

Lysozyme (LZM) is a natural anti-bacterial protein that is found in the saliva, tears and milk of all mammals including humans. Its anti-bacterial properties result from the ability to cleave bacterial cell walls, causing bacterial death. The current study was conducted to investigate the effects of dietary LZM on fecal microbial composition and variation in metabolites in sow. The addition of LZM decreased the fecal short-chain fatty acids (SCFAs). Zonulin and endotoxin in the serum, and feces, were decreased with lysozyme supplementation. Furthermore, fecal concentrations of lipocalin-2 and the pro-inflammatory cytokine TNF-α were also decreased while the anti-inflammatory cytokine IL-10 was increased by lysozyme supplementation. 16S rRNA gene sequencing of the V3-V4 region suggested that fecal microbial levels changed at different taxonomic levels with the addition of LZM. Representative changes included the reduction of diversity between sows, decreased Bacteroidetes, Actinobacteria, Tenericutes and Spirochaetes during lactation as well as an increase in Lactobacillus. These findings suggest that dietary lysozyme supplementation from late gestation to lactation promote microbial changes, which would potentially be the mechanisms by which maternal metabolites and inflammatory status was altered after LZM supplementation.Microbial diversity in sows' feces. A total of 54 fecal samples were subjected to 16S rRNA gene sequencing. Average raw reads, average effective tags and average operational taxonomic units (OTUs) for each treatment were shown in Supplementary Table S1. A set of 1,272 OTUs existed in all treatments and were thus defined as core OTUs (Fig. S2). These comprised 71.5% of the total number of OTUs, whereas 44,37,52, 135,47, 105, 72,56 and 57 OTUs were uniquely identified at Con. d1, LZM 150 d1, LZM 300 d1, Con. d7, LZM 150 d7, LZM 300 d7, Con. d21, LZM 150 d21 and LZM 300 d21, respectively (Fig. S2). To determine the bacterial diversity, the alpha and beta diversity of the fecal microbiota were assessed. We then compared the richness (observed species and Chao 1 index) and diversity (Shannon index) indices for the alpha diversity. As demonstrated in Table 2, LZM 300 diets decreased the observed species and Chao 1 index at day 21 of lactation. The lactation stage had a significant effect on fecal microbial community richness, with a Chao 1 index on day 1 of lactation being lower than at other stages (P < 0.01). A decrease in Shannon index (fecal microbial community diversity) with LZM supplementation at 300 mg/kg was found on day 7 and 21 of lactation (P < 0.01). For the analysis of beta diversity, the relationships among Control, LZM 150 and LZM 300 on day 1, 7 and 21 of lactation in the gut microbiome were examined by principal component analysis. The gut microbiota of sows showed obvious segregation in the different treatments, especially in treatment LZM300 d21 (Fig. 3) based on weighted UniFrac distance. The PERMANOVA analysis found that the bacterial community structure was significantly (P < 0...

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