This study was conducted to investigate the effects of feeding oregano essential oil, butyrate, and its mixture on the intestinal microbial diversity of calves. A completely randomized experimental design was used. Sixty-four healthy neonatal Holstein female calves with birth weight ≥ 35 kg were randomly divided into one control and three treatments (16 calves per group). The control group was fed normally, and the treatment group was fed oregano essential oil, butyrate, and their mixture, respectively. The experiment lasted for 70 days, and the lactation period lasted for 56 days. On days 55 and 70, rectal fecal samples from five calves were collected from each group for 16S rRNA amplification and sequencing. The results showed as follows: (1) the three treatments had no significant effects on the intestinal microbial community diversity, community uniformity, and community pedigree diversity of calves (p > 0.05). (2) At the phylum level, Firmicutes, Bacteroidota, Spriochatetota, Actinobacteriota, Firmicutes, and Bacteroidota gates of the main bacteria were detected in feces. (3) At the genus level, the top ten species with relative abundance detected are: norank_ F_Muribaaculaceae, Ruminococcus, unclassified_ F_ Lachnospiraceae, UCG-005, Prevotelaceae_NK3B31_Group, Prevotella, Bacteroides, Rikenellaceae_RC9_Gut_Group, and Faecalibacterium, Alloprevotella. (4) LEfSe analysis results show that the species with significant differences in the control group were f__Lachnospiraceae, o__Lachnospirales, o__Coriobacteriales, and c__Coriobacteriia, g__Megasphaera; in the essential oil group were g__Lachnospiraceae_AC2044_group, o__Izemoplasmatales, g__norank_f__norank_o__Izemoplasmatales, and f__norank_o__Izemoplasmatales; in the sodium butyrate group were g__Lachnospiraceae_NK4A136_group, and g__Sharpea, g__Fournierella; in the mixed group were g__Flavonifractor, and g__UBA1819. (5) The functional prediction analysis of calf gut microbes, found on the KEGG pathway2, shows that essential oil significantly improved membrane transport, Sodium butyrate inhibits lipid metabolism and improves the body's resistance to disease. (p < 0.05). (6) The effects of each treatment on the intestinal microbial structure of calves did not last for 14 days after the treatment was stopped. In conclusion, the addition of oregano essential oil, butyrate, and its mixtures to milk fed to calves can modulate the microbial structure, and it is recommended that oregano essential oil and butyrate be used separately, as a mixture of the two can increase the rate of diarrhea in calves.
Keywords: fecal micro-organisms; growth performance; oregano essential oil; sodium butyrate.