Increased dietary zinc oxide changes the bacterial core and enterobacterial composition in the ileum of piglets

This study was conducted to investigate the effects of increased dietary ZnO on the bacterial core and enterobacterial composition in the small intestine of piglets that were fed diets containing a total of 124 or 3,042 mg of Zn per kilogram of diet, respectively. Zinc was supplemented to the basal diet as ZnO. Bacterial 16S rRNA genes of ileal DNA extracts were PCR-amplified with 2 bar-coded primer sets and sequenced by 454 pyrosequencing. The bacterial core species were calculated from the relative abundances of reads present in 5 of 6 samples per group and at a minimum of 5 sequences per sample. The reference database SILVA was used to assign sequence reads at an alignment minimum of 200 bases and 100% identity. Lactic acid bacteria dominated the bacterial core, but showed diverse responses to dietary ZnO. Of the dominant Lactobacillus spp., Lactobacillus reuteri was reduced due to increased dietary ZnO (44.7 vs. 17.9%; P=0.042), but L. amylovorus was not influenced. However, the changes of relative abundances of other lactic acid bacteria were more noteworthy; Weissella cibaria (10.7 vs. 23.0%; P=0.006), W. confusa (10.0 vs. 22.4%; P=0.037), Leuconostoc citreum (6.5 vs. 14.8%; P=0.009), Streptococcus equinus (0.14 vs. 1.0%; P=0.044), and S. lutetiensis (0.01 vs. 0.11%; P=0.016) increased in relative abundance. Nonlactic acid bacteria that were influenced by increased dietary ZnO included the strict anaerobic species, Sarcina ventriculi, which showed a strong numerical decrease in relative abundance (14.6 vs. 5.1%). Species of the Enterobacteriaceae increased their relative abundance, as well as species diversity, in the high dietary ZnO experimental group. Bacterial diversity indices were increased due to increased dietary ZnO (P < 0.05), which was traced back to the increase of sequences from subdominant species. Increased dietary ZnO led to an increase of less prominent species and, thus, had a major impact on the bacterial composition and diversity in piglets. This effect may help to stabilize the intestinal microbiota in the sensitive postweaning period.