Additional file 5: Database 1. of GIS-supported epidemiological analysis on canine Angiostrongylus vasorum and Crenosoma vulpis infections in Germany

The data supporting the findings of present study. (CSV 2996 kb

Additional file 3: Table S1. of GIS-supported epidemiological analysis on canine Angiostrongylus vasorum and Crenosoma vulpis infections in Germany

Univariable statistical analysis of associations several independent variables and infection with A. vasorum or C. vulpis. (DOCX 16 kb

Additional file 2: Figure S2. of GIS-supported epidemiological analysis on canine Angiostrongylus vasorum and Crenosoma vulpis infections in Germany

Map presenting DLM factors defined by the final model together with A. vasorum- and C. vulpis - positive cases. (TIFF 12806 kb

Additional file 4: Table S2. of GIS-supported epidemiological analysis on canine Angiostrongylus vasorum and Crenosoma vulpis infections in Germany

Results of the full multivariable logistic regression model for A. vasorum and C. vulpis. (DOCX 14 kb

Supplementary Material_Version 2

<p>Cp values retrieved from the endogenous quality control (18S rRNA gene) as well as Cp values for the detection of FCoV RNA (7b gene).</p

In <i>Toxoplasma gondii</i> type I-infected cats reactions against type I, I/II and I/III specific peptides are strongest and are overrepresented in number.

<div><p>Intensities (MSIVs) by which clonal type I-infected cats reacted with individual peptides were analyzed using ANOVA and the Least Significant Difference (LSD)-Post-Hoc-Test (A). Whiskers represent 95% confidence intervals of the means of MSIV (bars). The differences between the means of MSIVs were regarded as statistically significant, when they were equal or higher than the LSD values. Different letters above the whiskers indicate significant differences between the mean intensities in the LSD-Post-Hoc-Test. </p> <p>To evaluate whether positive or negative serum reactions against clonal type-specific peptide cohorts were over- or underrepresented in cats infected with <i>T. gondii</i> clonal type I, a log-linear model analysis was used and the results presented in a mosaic plot (B). The size of each box in the mosaic plot corresponds to the observed frequencies of positive (Pos) and negative (Neg) peptide reactions as well as the number of analyzed peptides within each peptide cohort. Pearson residuals represent standardized deviations of observed from expected values. The Pearson residuals 0-2 with solid blue line indicate that the number of positive or negative reactions is higher, but not statistically significantly higher than expected (Pearson chi-squared p-value < 0.1). Blue scale shadings suggest the statistically significant rejection of the null hypothesis, i.e. overrepresentation of reactions against particular peptide groups (Pearson residuals (>2), Pearson chi-squared p-value < 0.05). Dashed red lines indicate an underrepresentation of positive or negative peptide reactions which is not statistically significant. Red scale shadings suggest a statistically significant rejection of the null hypothesis, i.e. underrepresentation of peptide reactions within the analyzed peptide group (Pearson residuals (<-2) Pearson chi-squared p-value, 0.05).</p></div

In naturally <i>Toxoplasma gondii</i> seropositive cats strongest reactions were observed against type II and I/II-specific peptides and reactions against these peptides were overrepresented.

<div><p>Intensities (MSIVs) by which naturally infected cats reacted with individual peptides were analyzed using ANOVA and the Least Significant Difference (LSD)-Post-Hoc-Test (A). To evaluate whether positive or negative serum reactions against clonal type-specific peptide cohorts were over- or underrepresented in naturally <i>T. gondii</i>-infected cats, a log-linear model analysis was carried out and results presented as mosaic plot (B).</p> <p>Detailed explanations of [A] and [B] are already given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080213#pone-0080213-g001" target="_blank">Figure 1</a>. </p></div

In cats infected with non-canonical <i>Toxoplasma gondii</i> strongest reactions were observed against type I/III specific peptides and the number of reactions against these peptides were overrepresented.

<div><p>Intensities (MSIVs) by which non-canonical type-infected cats reacted with individual peptides were analyzed using ANOVA and the Least Significant Difference (LSD)-Post-Hoc-Test (A). To evaluate whether positive or negative serum reactions against clonal type-specific peptide cohorts were over- or underrepresented in cats infected with atypical <i>T. gondii</i>, a log-linear model analysis was used and the results presented in a mosaic plot (B).</p> <p>Detailed explanations of [A] and [B] are already given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080213#pone-0080213-g001" target="_blank">Figure 1</a>. </p></div

Naturally <i>Toxoplasma gondii</i> seropositive cats and cats with a known clonal type II infection recognized similar peptide patterns.

<p>To explore whether there are particular patterns of anti-peptide reactions among all serologically positive <i>T. gondii</i> cat sera, a XY-fused Selforganizing Kohonen Network analysis (XYF-SKN) was performed. Figure (A) presents the clustering of various groups of <i>T. gondii</i> positive cat sera in a Y-map. Most of the sera derived from cats infected with non-canonical <i>T. gondii</i> types (A), infected by clonal types I, II or III (I, II, or III), or naturally infected cats (N) clustered in the grid nodes <i>1</i> to <i>8</i> either together (type III- and atypical type-infected [node <i>4</i>] as well as the type II and naturally infected cats [nodes <i>1</i> to <i>3</i> and <i>6</i> to <i>8</i>]) or separately (cats infected with type I <i>T. gondii</i> [node <i>5</i>]). Three cat groups were predicted using XYF-SKN, e.g. a cat group infected with non-canonical and type III <i>T. gondii</i> strains (<i>A</i>), a clonal type I infected group (<i>I</i>) and a naturally-infected cat group (<i>N</i>). Predicted cat groups are presented by different background colours of grid nodes, e. g <b><i>A</i></b> by red, <b><i>I</i></b> by green and <b><i>N</i></b> by blue. Figure (B) shows the number of group-specific sera within predicted clusters (<i>A</i>, <i>I</i> and <i>N</i>), further confirming that sera of naturally infected cats clustered mainly with type II reference sera. Figure (C) shows the number of group-specific sera within eight grid nodes in Figure (A).</p

In a <i>Toxoplasma gondii</i> type III-infected cat type III and I/III specific peptides were recognized strongest and were overrepresented in number.

<div><p>Intensities (MSIVs) by which clonal type III-infected cats reacted with individual peptides were analyzed using ANOVA and the Least Significant Difference (LSD)-Post-Hoc-Test (A). To evaluate whether positive or negative serum reactions against clonal type-specific peptide cohorts were over- or underrepresented in a cat infected with <i>T. gondii</i> clonal type III, a log-linear model analysis was used and the results presented in a mosaic plot (B).</p> <p>Detailed explanations of [A] and [B] are already given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080213#pone-0080213-g001" target="_blank">Figure 1</a>. </p></div