Immunolocalization of the S3Pvac peptides in <i>E. granulosus</i> and <i>E. multilocularis</i>.

<p>Sections of larval specimens reveal a strong binding to the tegument (<b>T</b>) as well as in the parenchyma (<b>P</b>) of <i>E. multilocularis</i> and in tegument of <i>E. granulosus</i>. (<b>A</b>) Control labeled with rabbit pre immune serum. (<b>B</b>) Positive control labeled with polyclonal sera from rabbit immunized with a total extract of <i>Taenia crassiceps</i> ORF cysticerci.</p

Sequences used to model the 3D structure of KETc1, GK1 and KETc12.

1<p>GK1 and KETc1 3D models include more than just the peptide sequence because the HMMSTR-server requires sequences of at least 20 amino acids length. Upper case marks the sequences of peptides while lower caps mark the preceding sequence to them.</p>2<p>There is no “complete” sequence for any of the strains of <i>T. crassiceps.</i> However, considering the strong similarity between the <i>Taenia</i> species we decided to construct a hypothetical sequence for each strain assuming that the preceding regions to KETc1 in <i>T. crassiceps</i> spp. are identical to those preceding the peptide in the other <i>Taenia</i> species.</p

Multiple sequence alignment GK1 peptides.

<p>The DNA and amino acid sequences of GK1 in several species of the Taeniidae family were aligned. GK1 alignments of amino acids (A) and cDNA (B) are shown in the upper part of the figure; the region encoding the actual peptides is marked with a double-headed arrow on the top of the alignment. On every alignment, boxes corresponding to the regions subjected to dN/dS-ratio analysis are marked accordingly. Models of the 3D structure of the GK1 peptide are shown in panel C. The model in red corresponds to <i>T. solium</i>, <i>T. saginata</i> and <i>E. granulosus</i>' GK1 and in purple to <i>T. crassiceps</i> ORF and WFU and <i>E. multilocularis</i>. A very good fitting between the two structures is shown as expected from the strong similarity of the amino acid sequence.</p

S3Pvac peptides were detected in the tegument of <i>Taenia</i> tapeworms and <i>T. solium</i> eggs.

<p>No reaction was detected in control sections, for which naive rabbit serum was used in place of the specific polyclonal rabbit serum. <b>S.</b>- Sucker; <b>Sc.</b> Scolex, <b>MP.</b>- Medullar parenchyma, <b>N.</b>- Neck; <b>DC.</b>- Distal cytoplasm region; <b>PE.</b>- perinuclear cytoplasm region. <b>O.</b>- oncospheres.</p

Primers used for PCR amplification of the KETc1 and GK-1 (KETc7) homologous sequences in <i>T. crassiceps</i> ORF, <i>T. crassiceps</i> WFU, <i>T. solium</i>, <i>T. saginata</i>, <i>E. multilocularis</i>, and <i>E. granulosus.</i>

<p>Primers used for PCR amplification of the KETc1 and GK-1 (KETc7) homologous sequences in <i>T. crassiceps</i> ORF, <i>T. crassiceps</i> WFU, <i>T. solium</i>, <i>T. saginata</i>, <i>E. multilocularis</i>, and <i>E. granulosus.</i></p

Multiple sequence alignment of KETc1 peptides.

<p>The DNA and amino acid sequences of KETc1 of several species of the Taeniidae family were aligned. KETc1 alignments of amino acids (A) and cDNA (B) are shown in the upper part of the figure. On every alignment, boxes corresponding to the regions subjected to dN/dS-ratio analysis are marked accordingly. Panel C and D shows the 3D models of the complete and truncated version of KETc1. Panel D depicts the models corresponding to <i>T. solium</i> (red), <i>T. crassiceps</i> ORF and WFU (pink), <i>T. saginata</i> (light green) and <i>Echinococcus</i> group (dark green). Structures are aligned with very good general agreement. Panel D shows the alignment of the truncated KETc1 peptide where the yellow model corresponds to <i>T. solium</i> and <i>T. saginata</i>; blue corresponds to <i>T. crassiceps</i> ORF and WFU and brown to the <i>Echinococcus</i> group.</p

Immunolocalization of the S3Pvac peptides in larval specimens of the four Taeniidae families.

<p>Metacestode cryostat parasite sections were incubated with a rabbit pre-immune serum (A) or with the three different rabbit antibodies (anti-GK1, anti-KETc1, anti-KETc12) and developed with biotinylated goat anti-rabbit IgG plus streptavidin peroxidase conjugate and counterstained with hematoxilin. The arrows indicate the areas in the metacestode where the peptides were localized. 10X (A and C), 40X (B and D). Tegument (<b>T</b>), parenchyma (<b>P</b>), tegument of the spiral canal (<b>TSC</b>), parenchymal folds (<b>PF</b>), microthrix (<b>MT</b>), parenchyma (<b>P</b>). The three peptides are present in metacestode stage of four cestodes analyzed.</p