Gut microbial alterations in neonatal jaundice pre- and post-treatment

Abstract

Neonatal jaundice is a common disease that affects up to 60% of newborns. Herein, we performed a comparative analysis of the gut microbiome in neonatal jaundice and non-neonatal jaundice infants (NJIs) and identified gut microbial alterations in neonatal jaundice pre- and post-treatment. We prospectively collected 232 fecal samples from 51 infants at five time points (0, 1, 3, 6, and 12 months). Finally, 114 samples from 6 NJIs and 19 non-NJI completed MiSeq sequencing and analysis. We characterized the gut microbiome and identified microbial differences and gene functions. Meconium microbial diversity from NJI was decreased compared with that from non-NJI. The genus Gemella was decreased in NJI versus non-NJI. Eleven predicted microbial functions, including fructose 1,6-bisphosphatase III and pyruvate carboxylase subunit B, decreased, while three functions, including acetyl-CoA acyltransferase, increased in NJI. After treatments, the microbial community presented significant alteration-based β diversity. The phyla Firmicutes and Actinobacteria were increased, while Proteobacteria and Fusobacteria were decreased. Microbial alterations were also analyzed between 6 recovered NJI and 19 non-NJI. The gut microbiota was unique in the meconium microbiome from NJI, implying that early gut microbiome intervention could be promising for the management of neonatal jaundice. Alterations of gut microbiota from NJI can be of great value to bolster evidence-based prevention against 'bacterial dysbiosis'.

Keywords: Gut microbiota; MiSeq sequencing; Neonatal jaundice; Treatment.

Figure 1. Study design and flow diagram

A total of 232 fecal samples from 15 NJI and 36 non-NJI were collected. After a strict pathologic diagnosis and exclusion process, the remaining samples were used for DNA extraction, 16S rRNA sequencing and data quality control. Finally, 6 NJI with treatment and 19 non-NJI were utilized for bioinformatics analysis.

Figure 2. Altered meconium microbiota composition in NJI

(A) Microbial α diversity decreased in NJI, as shown by the ACE estimator. (B) Shared and unique genera among NJI-0M, non-NJI-0M, NJI-1M, and non-NJI-1M. Overall diversity was calculated using unweighted UniFrac by PCoA (C) and NMDS (D), indicating a separation of samples between NJI and non-NJI. Fecal microbiota composition at the phylum level (E) and genus level (F) between NJI and non-NJI.

Figure 3. Identification of specific bacterial taxa and microbial functions associated with neonatal jaundice

(A) The greatest differences in taxa between NJI and non-NJI are presented according to the LDA scores (log10). (B) Differences in gut microbial functions between NJI and non-NJI based on the LDA scores (log10).

Figure 4. Gut microbial differences of infants between pre-treatment (NJI-0M) and post-treatment (NJI-1M)

Overall diversity was calculated using unweighted UniFrac by PCoA (A) and NMDS (B), indicating a separation of samples between pre-treatment and post-treatment. (C) PCoA based on the weighted UniFrac distance in NJI with treatment to assess the microbial distribution among 0, 1, 3, 6, and 12 months, indicating that samples tended to be uniform at 0 and 12 months, and samples are most heterogeneous at 1–6 months. Fecal microbiota composition at the phylum level (D) and genus level (E) between pre-treatment and post-treatment. (F) Compared with pre-treatment, two phyla were significantly increased, while two phyla were significantly decreased post-treatment (all P<0.05). (G) One genus was increased whereas one genus was decreased in pre-treatment versus post-treatment (all P<0.05). Abbreviations: M0, 0 month; M1, 1 month; M3, 3 months; M6, 6 months; M12, 12 months.

Figure 5. Identification of specific bacterial taxa and microbial functions between pre-treatment (0 month) and post-treatment (1 month)

(A) The greatest differences in taxa between pre-treatment and post-treatment are presented according to the LDA scores (log10). (B) Differences in gut microbial functions between pre-treatment and post-treatment based on the LDA scores (log10). Abbreviations: M0, 0 month; M1, 1 month.

Figure 6. Gut microbial alterations of infants between recovered NJI and non-NJI

(A) PCoA based on the weighted UniFrac distance between recovered NJI and non-NJI at 0, 1, 3, 6, and 12 months, indicating that the microbial communities became more uniform over time. Fecal microbiota composition at the phylum level (B) and genus level (C) between recovered NJI and non-NJI. (D) Compared with recovered NJI, one phylum was significantly decreased in non-NJI (P<0.05). (E) Three genera were increased in non-NJI versus recovered NJI (all P<0.05). Abbreviations: M0, 0 month; M1, 1 month; M3, 3 months; M6, 6 months; M12, 12 months.

Figure 7. Identification of specific bacterial taxa and microbial functions between recovered NJI and non-NJI

(A) The greatest differences in taxa between recovered NJI and non-NJI are presented according to the LDA scores (log10). (B) Differences in gut microbial functions between recovered NJI and non-NJI based on the LDA scores (log10).