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. 2024 Dec 28;14(1):30902.
doi: 10.1038/s41598-024-81227-y.

Temporal dynamics in the composition of metabolically active bacteria and fungi in the ileo-cecal lymph nodes of suckling and newly weaned piglets

Barbara U Metzler-Zebeli  1   2 , Frederike Lerch  3   4 , Fitra Yosi  3   4   5 , Julia C Vötterl  3   4 , Simone Koger  4   6 , Jutamat Klinsoda  3   7
Affiliations

Temporal dynamics in the composition of metabolically active bacteria and fungi in the ileo-cecal lymph nodes of suckling and newly weaned piglets

Barbara U Metzler-Zebeli et al. Sci Rep. .

Abstract

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.

Keywords: Bacteriome; Ileo-cecal lymph nodes; Microbial translocation; Mycobiome; Neonatal piglets; Weaning.

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Conflict of interest statement

Declarations. Ethical approval: All procedures involving animal handling and treatment were approved by the institutional ethics committee of the University of Veterinary Medicine Vienna and the National authority according to the Law for Animal Experiments, Tierversuchsgesetz (BMWFW- 68.205/00936- V3b/2019). The pig experiment was performed in accordance with the university and national regulations and complied to the ARRIVE guidelines. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Bacterial 16 S rRNA and fungal 26 S rRNA gene copies in ileo-cecal lymph nodes. (A) total bacteria, and (B) total fungi. One piglet group [sow milk group; n = 10 per day of life (DoL)] drank only sow milk during the suckling phase, the other piglet group [creep feed group; n = 10 per DoL] received additional creep feed from DoL10 until weaning which took place on DoL28. No creep feed effects and interactive effects of DoL and creep feed on total microbial abundances (p > 0.1).
Fig. 2
Fig. 2
Species richness and alpha-diversity of bacterial (blue) and fungal (orange) communities in ileo-cecal lymph nodes. Age-related differences for (A) Observed amplicon sequence variant (ASV) index, (B) Shannon index, and (C) Simpson index in suckling and newly weaned piglets. One piglet group [sow milk group; n = 10 per day of life (DoL)] drank only sow milk during the suckling phase, the other piglet group [creep feed group; n = 10 per DoL] received additional creep feed from DoL10 until weaning which took place on DoL28. No creep feed effects and interactive effects of DoL and creep feed on diversity indices (p > 0.1).
Fig. 3
Fig. 3
Non-metric multidimensional scaling (NMDS) plot of pairwise Bray-Curtis dissimilarities. (A) Bacteria (stress level = 0.0802), and (B) fungi (stress level = 0.0910) in ileo-cecal lymph nodes of suckling and newly weaned piglets. Ellipses represent the standard deviation. Days of life are indicated by numbers (7, 14, 21, 28, 31 and 35) in the center of the ellipses.
Fig. 4
Fig. 4
Relative abundances of the 20 most abundant bacterial and fungal genera in ileo-cecal lymph nodes. (A) Bacterial genera, and (B) fungal genera based on 16 S rRNA and ITS2 gene sequencing, respectively, in suckling and newly weaned piglets. One piglet group [sow milk group; n = 10 per day of life (DoL)] drank only sow milk during the suckling phase, the other piglet group [creep feed group; n = 10 per DoL] received additional creep feed from DoL10 until weaning which took place on DoL28. Effects (adjusted p < 0.05) and trends (adjusted p < 0.10) of DoL is indicated by ‘*’ and ‘#’, respectively.
Fig. 5
Fig. 5
Age- and creep feed-related effects on genera. Relative abundance of (A) Bacteroides, (B) Prevotellaceae NK3B31 group, (C) Alternaria, and (D) Trichoderma in the ileo-cecal lymph nodes of suckling and newly weaned piglets. One piglet group [sow milk group; n = 10 per day of life (DoL)] drank only sow milk during the suckling phase, the other piglet group (creep feed group; n = 10 per DoL) received additional creep feed from DoL10 until weaning which took place on DoL28. A-B represent bacterial genera, whereas C-D represent fungal genera.
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