Significance of co-infection patterns between selected operational taxonomic units (OTUs).

<p>Only data from the Georgia tick DNA sample collection is shown. The upper triangle represents <i>p</i>-values of positive associations (codetection in the same sample) between OTUs and the lower triangle the <i>p</i>-values of negative associations. Black cells were not evaluated due to low expected co-occurrence (<1) between the OTUs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102130#pone.0102130-Veech1" target="_blank">[49]</a>.</p

Life stage and geographic origin of <i>Amblyomma americanum</i> ticks used in bacterial community analyses.

§<p>Nymphs collected in July 2010; adults in July 2010 and May 2011.</p><p>*Nymphs collected in August 2010; adults in May 2011.</p><p>NT = Not tested.</p

Bacterial operational taxonomic unit (OTU) sequence abundance detected in tick samples.

<p>OTUs present in greater than one third of all tick samples were considered broadly distributed. OTUs present at a mean abundance ≥1% of the community (i.e. 10 sequences) were considered locally abundant. Sequence abundance values are slightly offset to reveal overlapping OTUs.</p

Multivariate analysis of the effect of sex and life stage on the relatedness of <i>Amblyomma americanum</i> bacterial communities.

<p>Tick DNAs from the archival collection (n = 27) were older, collected in a different geographical region, and processed using different protocols than tick DNAs from the Georgia collection (n = 104).</p><p>*Corrected for multiple comparisons using Holm’s method; α  = 0.05.</p

Non-metric multi-dimensional scaling (NMDS) of a Bray-Curtis distance matrix describing archival <i>Amblyomma americanum</i> bacterial communities.

<p>Each point symbolizes a single tick’s community (n = 27); some points may overlap completely. Point and ellipse colors indicate life stage; ellipses represent 95% confidence intervals around life stage centroids. Non-overlapping centroids are considered significantly different at α = 0.05. R² values in the upper left corner of plots describe the amount of variation in the data set explained by the groupings. The stress value is a measure of the disagreement between the rank order in the original data set and that in the NMDS (lower numbers indicate better agreement). Numbers indicate the species scores for the plot as follows: (1) <i>Coxiella</i>, (2) <i>Rickettsia</i>, (3) <i>Midichloria</i> 1, (4) <i>Borrelia</i>, (5) <i>Francisella</i>, (6) <i>Midichloria</i> 2, (7) <i>Ehrlichia</i>, (8) Enterobacteriaceae, (9) <i>Pseudomonas</i> 1, (10) Burkholderiaceae, (11) Acidobacteria Gp1, (12) <i>Pseudomonas</i> 2, (13) Rhizobiales 2, (14) <i>Bacillus</i>, (15) Acetobacteraceae 1, (16) Acetobacteraceae 2, (17) Bacteria 1, (18) Bacteria 2.</p

Non-metric multi-dimensional scaling (NMDS) of a Bray-Curtis distance matrix describing Georgian <i>Amblyomma americanum</i> bacterial communities.

<p>Each point symbolizes a single tick’s community (n = 104); some points may overlap completely. Point and ellipse colors indicate life stage; ellipses represent 95% confidence intervals around life stage centroids. Non-overlapping centroids are considered significantly different at α = 0.05. R² values in the upper left corner of plots describe the amount of variation in the data set explained by the groupings. The stress value given is a measure of the disagreement between the rank order in the original data set and that in the NMDS (lower numbers indicate better agreement). Brown numbers indicate the species scores for select OTUs as follows: (1) <i>Coxiella</i>, (2) <i>Rickettsia</i>, (3) <i>Midichloria</i> 1, (4) <i>Borrelia</i>, (5) <i>Ehrlichia</i>, (6) Bacillales 1, (7) <i>Wolbachia</i>, (8) Rhizobiales 1, (9) <i>Rhodobaca</i>, (10) Bacillales 2, (11) <i>Nitriliruptor</i>, (12) Bacillales 3.</p

Plasmid based protein composition of <i>B</i>. <i>mayonii</i> in comparison to 4 other Bbsl genospecies and 1 RF <i>Borrelia</i> species.

<p>The mean amino acid identity is indicated for proteins present on each of the 14 <i>B</i>. <i>mayonii</i> plasmids. Comparisons are to protein databases constructed for <i>B</i>. <i>burgdorferi</i> (B.b.s.s.; green), <i>B</i>. <i>garinii</i> (B.g.; blue), <i>B</i>. <i>afzelii</i> (B. a.; red), <i>B</i>. <i>bissettii</i> (B.b.; yellow), and <i>B</i>. <i>miyamotoi</i> (B.m.; grey).</p

Comparison of <i>B</i>. <i>burgdorferi</i> B31 and <i>B</i>. <i>mayonii</i> lp54.

<p>Heat map of nucleotide identity between lp54 plasmids of <i>B</i>. <i>burgdorferi</i> B31 (top) and <i>B</i>. <i>mayonii</i> MN14-1420 (bottom). Nucleotide identity along the entire lp54 plasmids is displayed as colored blocks (Yellow = highest, Blue = lowest). Orientation of arrows indicates gene orientation on the forward or reverse DNA strand. The variable PF54 gene array is at the 3’ end of the plasmid. Specific gene loci discussed in text are indicated.</p

<i>B</i>. <i>mayonii</i> proteins with greatest sequence diversity as compared to 4 other Bbsl genospecies and 1 RF Borrelia species.

<p>Bar graph depicting <i>B</i>. <i>mayonii</i> proteins greater than 100 amino acids with less than 60% amino acid identity to other Bbsl proteins. Amino acid identity is shown with respect to homologs present in <i>B</i>. <i>burgdorferi</i> (B.b.s.s.; green), <i>B</i>. <i>garinii</i> (B.g.; blue), <i>B</i>. <i>afzelii</i> (B. a.; red), <i>B</i>. <i>bissettii</i> (B.b.; yellow) and <i>B</i>. <i>miyamotoi</i> (B.m.; grey).</p

Phylogenetic analysis of <i>B</i>. <i>mayonii</i> based on a ~6.5 kbp region of cp26.

<p>The ~6.5 kbp region of cp26 encompasses open reading frames from BB_B14 through BB_B18 of <i>B</i>. <i>burgdorferi</i> B31. DNA sequences from 8 other Bbsl genospecies (22 strains) were retrieved from GenBank. Bootstrap values greater than 50% are shown. The scale bar corresponds to 0.02 substitutions per nucleotide position.</p