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PLoS One
2021 Apr 22;164:e0241026. doi: 10.1371/journal.pone.0241026.
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Surveillance of densoviruses and mesomycetozoans inhabiting grossly normal tissues of three Aotearoa New Zealand asteroid species.
Hewson I
,
Sewell MA
.
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Asteroid wasting events and mass mortality have occurred for over a century. We currently lack a fundamental understanding of the microbial ecology of asteroid disease, with disease investigations hindered by sparse information about the microorganisms associated with grossly normal specimens. We surveilled viruses and protists associated with grossly normal specimens of three asteroid species (Patiriella regularis, Stichaster australis, Coscinasterias muricata) on the North Island / Te Ika-a-Māui, Aotearoa New Zealand, using metagenomes prepared from virus and ribosome-sized material. We discovered several densovirus-like genome fragments in our RNA and DNA metagenomic libraries. Subsequent survey of their prevalence within populations by quantitative PCR (qPCR) demonstrated their occurrence in only a few (13%) specimens (n = 36). Survey of large and small subunit rRNAs in metagenomes revealed the presence of a mesomycete (most closely matching Ichthyosporea sp.). Survey of large subunit prevalence and load by qPCR revealed that it is widely detectable (80%) and present predominately in body wall tissues across all 3 species of asteroid. Our results raise interesting questions about the roles of these microbiome constituents in host ecology and pathogenesis under changing ocean conditions.
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33886557
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Fig 1. Sampled specimens of Stichaster australis (A-B), Coscinasterias muricata (C-D) and Patiriella regularis (E-F).Viral metagenomes were prepared from body wall (b) samples collected by biopsy punch. Additional specimens of gonad (g) and pyloric caeca (p) were collected for quantification of viral genotypes and the mesomycetozoan.
Fig 2. Contig map of densovirus-like genome fragments recovered from Coscinasterias muricata and Patiriella regularis viral metagenomes.The colors of arrows indicate densoviral gene, and the best match (by BLASTx against the non-redundant database at NCBI) along with e-value is indicated adjacent to each ORF. The black lines running through ORFs indicate total contig length. The numbers in brackets below each contig are the number of reads recruiting to the contig from the origin library.
Fig 3. Phylogenetic representation of Patiriella regularis and Coscinasterias muricata-associated densoviral genome fragments.The trees are based upon 170 amino acid (Non-Structural), 211 amino acid (Structural; middle), and 104 amino acid (Structural; bottom) alignments performed using the CLC Sequence Viewer version 8.0. The trees were constructed with neighbor joining and Jukes-Cantor distance, where bootstrap values (1000 reps) are indicated above nodes. Red labels indicate sequences obtained in this study, while green labels indicate sequences obtained from asteroids in other studies.
Fig 4. Phylogenetic representation of asteroid-associated 28S rRNA sequences in purified virus metagenomes.The tree was constructed by neighbor joining and based on an 849 nucleotide alignment of eukaryotic 28S rRNA. Shown are close matches by BLAST against the non-redundant database.
Fig 5. Mesomycetozoan 28S rRNA copies as determined by qPCR in asteroid tissues.a,b denotes significant difference (p < 0.025, Studentâs t-test with Bonferroni correction for 2 comparisons).
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