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Next, we used supervised tensor learning and classified MTBC strains into major lineages using spoligoype deletions and MIRU patterns.
As a way of validating multiple-biomarker tensors, we use them in a supervised learning model to predict major lineages using spoligotype deletions and MIRU.
In order to do this, we estimated FST coefficients with individuals nested in colonies and colonies nested in major lineages using a three-level hierarchy in the GDA program [ 55], for each pair of putative cryptic species.
In our experiments, we used spoligotype and MIRU as biomarkers and predicted the six major lineages using the same data as for the above unsupervised learning experiments combined into a single dataset.
We determined parameters of within-population variability for all samples and major lineages using Arlequin 3.5 [ 70], calculating the number of segregating sites and haplotypes, haplotype diversity (Hd), nucleotide diversity, and Fu's FS[ 71], an indicator of population expansion when it is negative and significant (p ≤ 0.02) [ 35, 70, 71] (Table 1).
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The historical effects of natural selection on the cytb protein were estimated with the software TreeSAAP using a phylogenetic reconstruction for 107 mammal taxa from all major mammalian lineages using 13 complete protein-coding mitochondrial gene sequences.
Similarly, a recent study of major snake lineages using 333 anchored phylogenomics loci and species-tree methodologies found low support values within lamprophiids, the family to which pseudoxyrhophiines belong [ 15].
We used the concatenated nucleotide sequence data for all six genes, the BI phylogenetic tree topology of Figure 1, and several fossil-based minimum age constraints to estimate divergence times for major holometabolous lineages using the relaxed clock BI method implemented in the programs Estbranches and Multidivtime [ 53].
We calculated DNA polymorphism statistics to summarize the genetic diversity of each gene and major mtDNA lineages using DnaSP v5.10.1 [ 54], and calculated the following statistics: number of haplotypes (H), haplotype diversity (Hd), Watterson's theta (θ w ), nucleotide diversity per site (Pi), average number of nucleotide differences between sequences (k), and number of segregating sites (S).
We examined four major differentiated intestinal epithelial lineages using established markers.
Isolation by distance was tested within each major mtDNA lineage using a Mantel test of correlation between genetic and geographic distance matrices implemented in the software package IBD [ 42].
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