Increasing evidence has indicated the ameliorative effect of milk replacer supplementation in ruminants for regulating their early growth and rumen development. However, it is still unclear whether milk replacer supplementation has a beneficial role in the growth performance and rumen microbiota of Yimeng black goats (YBGs). Therefore, this study was performed to investigate the effects of milk replacer on growth performance and rumen microbiota of YBGs. Our results revealed that milk replacer supplementation could significantly improve the growth performance of YBGs. Additionally, the results of alpha and beta diversities indicated that there was no significant difference in richness and diversity between the control and milk replacer-treated YBGs. At the phylum level, Bacteroidetes, Firmicutes, and Proteobacteria were the most dominant phyla in all the samples at different stages. Moreover, the YBGs treated with milk replacer possessed a higher abundance of Verrucomicrobia than that in the control YBGs, while the level of Actinobacteria was obviously decreased. It is noteworthy that the abundance of Proteobacteria in the control YBGs was higher than that in the YBGs supplemented with milk replacer throughout the experiment. At the level of genus, the differences in the richness between control and milk replacer supplement YBGs were gradually observed. Compared with the control YBGs, the proportion of Akkermansia, Veillonella, Anaerovibrio, Ruminococcaceae_NK4A214_group, Ruminococcus_1, and Ruminococcus_2 was increased in the YBGs treated with milk replacer, whereas Turicibacter was decreased. In conclusion, milk replacer supplementation may serve as a good applicant for ameliorating early YBGs development and rumen microbiota.

Growth differentiation factor 9 (GDF9) plays an important role in the early folliculogenesis of sheep. This study investigated the mRNA expression of ovine GDF9 in different tissues by real-time PCR. GDF9 exhibits significantly higher levels of expression (p < 0.01) in the ovary, relative to other tissues, indicating that its expression is tissue specific. To explore the regulatory mechanism of this tissue-specific expression, the methylation level of one CpG island (−1453 to −1854) of GDF9 promoter in ovary and heart was determined. In this region (−1987 to −1750), only the mC-4 site was present in the Sp4 binding site showed differential methylation between the heart and ovary; with increased (p < 0.01) methylation being observed in the heart. Additionally, the methylation level was negatively correlated with GDF9 mRNA expression (R = −0.75, p = 0.012), indicating that the methylation of this site plays an important role in transcriptional regulation of GDF9. The methylation effect of the mC-4 site was confirmed by using dual-luciferase. Site-directed mutation (methylation) of mC-4 site significantly reduced (p < 0.05) basal transcriptional activity of GDF9 promoter in oocytes. These results imply that methylation of GDF9 promoter CpG island mC-4 site may affect the binding of the Sp4 transcription factor to the GDF9 promoter region in sheep, thereby regulating GDF9 expression and resulting in a tissue-specific expression.

Yimeng black goat is one of the national breeds of geographical indication in China and is one of the key protected local livestock and poultry breeds of Shandong province. The complete mitochondrial genome sequence of Yimeng black goat was investigated in this study (GenBank accession no. MT134111). The mitogenome (16,640 bp) consisted of a non-coding control region (D-loop region), two ribosomal RNA (rRNA) genes, 13 protein-coding genes (PCGs), and 22 transfer RNA (tRNA) genes. The complete mitochondrial genome sequence and the neighbour-joining tree of the Yimeng black goat would contribute to further study in genetic mechanism and phylogenomic research of goats.