The early microbial colonization of the porcine gut is an important priming factor for gut and immune development. Nevertheless, little is known about the composition of microbes that translocate into the ileo-cecal lymph nodes (ICLN) in the neonatal phase. This study aimed to characterize age- and nutrition-related changes in the metabolically active bacterial and fungal composition of the ICLN in suckling and newly weaned piglets. Ten litters received only sow milk, while ten litters had access to creep feed from day of life (DoL) 10. Weaning occurred on DoL28. The ICLN were collected from 10 piglets/feeding group on each sampling day (DoL7, 14, 21, 28, 31 and 35) for RNA isolation, transcription into complementary DNA for 16 S rRNA and ITS2 amplicon sequencing. Age and weaning influenced the microbiome in the ICLN more than the nutrition during the suckling phase. Species richness and alpha-diversity of the bacterial but not fungal communities were increased on DoL7 and postweaning. Potential modes of action may have been linked to gut permeability at these ages and selective sampling by immune cells. Potential selective transfer of microbes may explain the dominance of Lactobacillus and Limosilactobacillus in the ICLN. Piglets that only drank sow milk comprised more Bacteroides in their ICLN on DoL35 compared to the creep fed piglets. Especially the role of fungi in the ICLN, including their mechanisms for translocation survival, needs further attention, as we detected metabolically active mold fungi and plant pathogens (e.g., Fusarium, Alternaria and Blumeria) in the ICLN.