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. 2024 Jul 2;14(1):15188.
doi: 10.1038/s41598-024-66097-8.

Characterising the Tasmanian devil (Sarcophilus harrisii) pouch microbiome in lactating and non-lactating females

Affiliations

Characterising the Tasmanian devil (Sarcophilus harrisii) pouch microbiome in lactating and non-lactating females

Lucy E Ockert et al. Sci Rep. .

Abstract

Wildlife harbour a diverse range of microorganisms that affect their health and development. Marsupials are born immunologically naïve and physiologically underdeveloped, with primary development occurring inside a pouch. Secretion of immunological compounds and antimicrobial peptides in the epithelial lining of the female's pouch, pouch young skin, and through the milk, are thought to boost the neonate's immune system and potentially alter the pouch skin microbiome. Here, using 16S rRNA amplicon sequencing, we characterised the Tasmanian devil pouch skin microbiome from 25 lactating and 30 non-lactating wild females to describe and compare across these reproductive stages. We found that the lactating pouch skin microbiome had significantly lower amplicon sequence variant richness and diversity than non-lactating pouches, however there was no overall dissimilarity in community structure between lactating and non-lactating pouches. The top five phyla were found to be consistent between both reproductive stages, with over 85% of the microbiome being comprised of Firmicutes, Proteobacteria, Fusobacteriota, Actinobacteriota, and Bacteroidota. The most abundant taxa remained consistent across all taxonomic ranks between lactating and non-lactating pouch types. This suggests that any potential immunological compounds or antimicrobial peptide secretions did not significantly influence the main community members. Of the more than 16,000 total identified amplicon sequence variants, 25 were recognised as differentially abundant between lactating and non-lactating pouches. It is proposed that the secretion of antimicrobial peptides in the pouch act to modulate these microbial communities. This study identifies candidate bacterial clades on which to test the activity of Tasmanian devil antimicrobial peptides and their role in pouch young protection, which in turn may lead to future therapeutic development for human diseases.

Keywords: Immunology; Marsupial; Microbiome; Reproduction; Skin.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Map of sites where pouch swab samples were collected from Tasmanian devils. Samples were collected from the following sites for microbial analysis; Narawntapu National Park (NP; N = 11), Stony Head (SH; N = 5), Fentonbury (FB; N = 18), Kempton (KP; N = 16), and Buckland (BL; N = 4). A breakdown of the number of samples collected for each sampling group can be found in Additional file 1: Table S2. Colour corresponds to month when fieldtrip took place, scale bar represents kilometres, and arrow indicates north. Graphic was created using QGIS (v 3.22) and Australian Bureau of Statistics Digital Boundary Files.
Figure 2
Figure 2
Taxonomy bar plots displaying the relative abundance of bacteria present in Tasmanian devil pouches across reproductive stages at three taxonomic levels; (A) Phylum, (B) Family, and (C) Genus. Each column represents one sample. Samples are organised according to reproductive stages; lactating and non-lactating. Location is indicated below the x-axis as follows; BL = Buckland, FB = Fentonbury, KP = Kempton, NP = Narawntapu National Park, and SH = Stony Head. Taxa with a median relative abundance of < 0.25% were merged into ‘Other’. Visualisation was performed using ggplot2 (v 3.4.2).
Figure 3
Figure 3
Alpha diversity metrics of pouch microbial compositions per reproductive status. Whisker box plots depict (A) Observed Amplicon Sequence Variant Richness, (B) Shannon Diversity, and (C) Faith’s Phylogenetic Diversity from lactating (purple; n = 25) and non-lactating (orange; n = 30) devils. Results of statistical analysis can be found in Additional file 1: Table S5. Visualisation was performed using ggplot2 (v 3.4.2).
Figure 4
Figure 4
Differences in microbial community composition observed between reproductive status and location. Nonmetric Multidimensional Scaling (NMDS) plots of (A) unweighted, and (B) weighted UniFrac distances. Colour represents reproductive status while shape indicates location. Results of statistical analysis can be found in Additional file 1: Table S6. Visualisation was performed using ggplot2 (v 3.4.2).
Figure 5
Figure 5
Differentially abundant amplicon sequence variants (ASVs) between reproductive groups as identified by DESeq2 analysis. Each point represents a single differentially abundant ASV, grouped according to genus. The comparison was made between lactating versus non-lactating pouches, hence a positive Log2 Fold Change indicates a higher prevalence in lactating pouches and a lower prevalence in non-lactating pouches. All ASVs visualised above were found to be significantly different between reproductive groups using a Kruskal–Wallis test. Visualisation was performed using ggplot2 (v 3.4.2).

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