Comparison – C.bairdi 20102558-2729 vs. 6129-403-26 NanoPore Taxonomic Assignments Using MEGAN6

After noticing that the initial MEGAN6 taxonomic assignments for our combined C.bairdi NanoPore data from 20200917 revealed a high number of bases assigned to E.canceri and Aquifex sp., I decided to explore the taxonomic breakdown of just the individual samples to see which of the samples was contributing to these taxonomic assignments most.

After completing the individual taxonomic assignments, I compared the two sets of assignments using MEGAN6 and generated this bar plot showing percentage of normalized base counts assigned to the following groups within each sample:

  • Aquifex sp.
  • Arthropoda
  • E.canceri
  • SAR (Supergroup within which Alveolata/Hematodinium sp. falls)
IMPORTANT!!!
  • The taxonomic makeup shown in these comparisons is only a comparison of bases assigned amongst the four taxa selected above. It is not a comparison of the full taxonomic makeup of the two samples. I will discuss the data shown here in that context.

20201002_cbai_nanopore_20102558-2729-Q7-vs-6129-403-26-Q7_megan-taxonomic-comparison-bar-plot

Comparison table:

Taxa 20102558-2729-Q7_base-counts 20102558-2729-Q7_base-counts(%) 6129-403-26-Q7_base-counts 6129-403-26-Q7_base-counts(%)
Aquifex sp. 221,823.00 10.25 199,287.06 10.43
Arthropoda 1,046,619.00 48.38 1,134,731.00 59.40
Enterospora canceri 889,082.00 41.10 561,754.19 29.41
Sar 5,855.00 0.27 14,582.56 0.76
TOTAL 2,163,379.00 1,910,354.81

Some observations:

  • Aquifex sp. account for nearly the same percentage of assignments in both samples.
  • Arthropoda makes up ~50% of assigned bases in the uninfected muscle sample (20102558-2729), but ~60% in the Hematodinium-infected hemolymph sample. (6129-403-26).
  • E.canceri makes up ~41% of assigned bases in the uninfected muscle sample (20102558-2729), but only ~30% in the Hematodinium-infected hemolymph sample.
  • SAR contributes a very small percentage to each of the two samples, but has ~2.8x the number of assigned bases. Additionally, as noted in the taxonomic assignment analysis of 20102558-2729-Q7 on 20200928, no bases are assigned to descendants of this Supergroup, whereas in the taxonomic analysis of 6129-403-26-Q7 on 20200928, there are bases assigned within the descendants of this Supergroup, down the level of Hematodinium sp. Genus.

Pretty interesting stuff!

I also briefly looked at the taxonomic assignments from all of our hemolymph RNAseq samples to see if if Aquifex sp. and/or E.canceri appear:

Interestingly, a high number of reads are assigned to E.canceri in all samples, but no reads are assigned to Aquifex sp.. Another observation is that a fair number of reads get assigned to Vibrio parahemolyticus, but very few number of NanoPore DNA bases get assigned to V.parahemolyticus.

Next up I think I might try to identify which contigs/scaffolds from the cbai_genome_v1.0 Flye assembly correspond to these taxa. The approach would be to create a BLAST database (DB) from the cbai_genome_v1.0.fasta (19MB). Then extract the NanoPore reads assigned to each of the taxa above, then BLAST them against the cbai_genome_v1.0 BLAST DB.

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