Distribution of mutations with regards to NA subtype.

<p>Mainly adopted from Orozovic et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089306#pone.0089306-Orozovic1" target="_blank">[26]</a>.</p>1)<p>Sum of all R118K mutations.</p>2)<p>Mutation unrelated to inhibitor resistance.</p>3)<p>Isolates that were not analyzed.</p>4)<p>Previously published values.</p

Sequence alignment of N2 reference, and three N1 sensitive mutants.

<p>The sequence interval between the 121^th and 149^th amino acid (marked with - - - ; N2 reference numbering) is excluded from alignment for practical reasons. The relevant mutation sites are indicated in red. (r) - reference sequence; Number within the parenthesis indicate code for virus isolates according <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089306#pone-0089306-t002" target="_blank">Table 2</a>. (3) - H1N1 R118K mutant; (4) - H1N1 R118K/D151N mutant; (14) - H6N1 R118K/D151N/D198N mutant. Cl.Co- ClustalW consensus sequence, “*” is used to indicate identical residues, “:” is used to indicate conserved substitution, “.” is used to indicate semi-conserved substitution, “ ” (empty space) is used to indicate dissimilar residues.</p

Performance of virus isolates in CM assay.

1)<p>Code for virus subtypes; *- mutations induced by OC (adopted from literature); without *- mutations induced by ZA (adopted from literature).</p>2)<p>Mutation unrelated to inhibitor resistance.</p>3)<p>F - framework residue; C - catalytic residue; WT- wild type.</p

IC₅₀ values of viral isolates analyzed by CM.

<p>Each bar shows mean ± SE IC₅₀ of three replicates. The x-axis shows the viral isolates. NxI and NxII represent wild types (WTs) and 1–14 represent mutants. For codes of enumeration, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089306#pone-0089306-t002" target="_blank">Table 2</a>. z – inhibition by ZA; o – inhibition by OC. On the y-axis, the IC₅₀ values are expressed as concentration of inhibitor in nM. All means were tested by one-way ANOVA and post-hoc Tukey tests. A) N1 subtype; B) N3 subtype; C) N6 subtype and D) N9 subtype.</p

Intracellular localization of viable and heat killed <i>C. jejuni</i> in <i>A. polyphaga</i>.

<p>The number of viable and heat killed <i>C. jejuni</i> found inside digestive- (circles) and non digestive (squares) vacuoles are shown after 1 h, 24 h, 48 h and 72 h of co-incubation. Viable <i>C. jejuni</i> were found inside non digestive vacuoles to a significantly larger extent than heat killed bacteria. This difference was not observed inside digestive vacuoles. Data for all time points are based on two to three independent experiments, except for viable <i>C. jejuni</i> and <i>A. polyphaga</i> at 72 h that was based on one experiment. Mean±95 confidence interval.</p

<i>A. polyphaga</i> trophozoites associated with <i>C. jejuni</i>.

<p>The fraction of <i>A. polyphaga</i> trophozoites associated with <i>C. jejuni</i> are shown as % of all counted trophozoites at 1 h, 24 h, 48 h and 72 h after infection. The trophozoites registered at each time point fall into one of the following categories: Trophozoites associated with viable <i>C. jejuni</i> only (squares), associated with heat killed <i>C. jejuni</i> only (circles), associated with both viable and heat killed <i>C. jejuni</i> (triangles) or associated with no <i>C. jejuni</i> (diamonds). Among the trophozoites associated with both viable and heat killed <i>C. jejuni,</i> the amount of viable <i>C. jejuni</i> were dominating at all time points except at 96 h. Data are based on three to five independent experiments for samples taken at 1 h–72 h and on one experiment for 96 h. Mean±95 confidence interval.</p

Viable and heat killed <i>C. jejuni</i> are taken up into different types of <i>A. polyphaga</i> vacuoles.

<p>(A) Live/Dead stained viable (green) <i>C. jejuni</i>, confined to tight vacuoles. (B) Live/Dead stained heat killed (red) <i>C. jejuni</i> residing in giant spacious vacuoles. (C) Vaculoes with CTC stained viable (red) <i>C. jejuni</i> do not co-localize with Alexa fluor-488 labeled dextran filled vacuoles (green). (D) Vaculoes with CTC stained heat killed (red) <i>C. jejuni</i> have taken up Alexa fluor-488 labeled dextran (green). (E) In contrast to non digestive vacuoles, giant digestive vacuoles contained smaller vesicles (arrow). Picture D and E are from the same amoeba. ZEISS Axioskop (Germany)×63, FI 450–490 (FT 510, LP 520) and a Nikon camera COOLPIX 995 was used for fluorescence images A, C, D and bright field image E. OLYMPUS BX 50 (Japan) ×100 and an OLYMPUS camera DP 50 was used for fluorescence image B.</p

Adhered/internalized viable and heat killed <i>C. jejuni</i> per <i>A. polyphaga</i> trophozoite.

<p>After 1-incubation the majority of amoeba associated viable <i>C. jejuni</i> were found on the surface of trophozoites but at 24 h and thereafter, the majority were found in intracellular vacuoles. The number of viable bacteria internalized into <i>A. polyphaga</i> trophozoites was significantly higher than for heat killed bacteria during the first 48 h. Data are based on three to five independent experiments for samples taken at 1 h–72 h and on one experiment for 96 h. Mean±95 confidence interval.</p

<i>E</i>. <i>coli</i> sequence types (ST), clonal complex and ESBL genotype(s) in isolated <i>E</i>. <i>coli</i> from Franklin's gulls in Canada.

<p><i>E</i>. <i>coli</i> sequence types (ST), clonal complex and ESBL genotype(s) in isolated <i>E</i>. <i>coli</i> from Franklin's gulls in Canada.</p

The number of <i>E</i>. <i>coli</i> (percent in brackets) with phenotypic resistance to ten antibiotics isolated from Franklin's gulls in Canada and Chile.

<p>The number of <i>E</i>. <i>coli</i> (percent in brackets) with phenotypic resistance to ten antibiotics isolated from Franklin's gulls in Canada and Chile.</p