Table 1.
Description of appendectomy sections based on age, pathological, and molecular results.
Fig 1.
Microscopic images of the cross-section of E. vermicularis in the appendix lumen in different magnifications (A, B, and C). Arrows illustrate the cephalic alae located bilateral and symmetrically in the sections. Images were drawn with camera lucida, Enterobius vermicularis cross-section in tissue sections emphasizing cephalic alae (D, E, and F). C1: Flattened and less than five somatic muscles in each quadrant in cross-section indicated platymyarian and meromyarian types of the somatic muscle.
Fig 2.
Electrophoresis of cox1 gene (378 bp) amplified fragments using nested PCR in paraffin tissue samples of patients with appendicitis due to E. vermicularis in Tehran and Hamedan provinces.
M:100 bp DNA Marker (Gene Ruler™, Fermentas); lane numbers 1–12: amplicons related to FFPE tissues; C+: positive control (the adult worm); NTC: no template control negative control.
Fig 3.
Median-Joining haplotype networks analysis of cox1 gene (378 bp) based on 12 isolates of E. vermicularis constructed using PopArt 1.7.
Six haplotypes were identified within the isolates in the current study. The details of the sequence(s) belonging to each haplotype are noted. Each circle represents a unique haplotype, n represents the number of sequences in each haplotype, and the hatch marks along the network branches indicate the number of mutations.
Table 2.
Details of statistics genetic diversity for 12 sequences of E. vermicularis in Iran.
Table 3.
Pairwise genetic distance matrix of cox1 sequences between 12 isolated Enterobius vermicularis.
Evolutionary analysis was conducted in MEGAX by using the Kimura 2-parameter model. The number of base substitutions per site from between sequences is shown.
Fig 4.
The amino acid alignment of mitochondrial cytochrome c oxidase 1 (cox1) gene of Iranian E. vermicularis haplotypes with marked changes in the amino acids sequence.
Dots denote identical amino acids to the top sequence.
Fig 5.
Phylogenetic relationships were inferred from 333 bp of cox1 gene sequences of Enterobius vermicularis isolates in this study (black circles) and others retrieved from GenBank.
The tree was constructed based on the maximum likelihood method in MEGAX with Ascaris suum as the outgroup. Bootstrap values (>50%) are shown at the nodes based on 1000 replicates. The scale bar indicates genetic distance. All the reference sequences of Enterobius vermicularis from GenBank are displayed with their accession number, the host origins, and the original countries. Types of E. vermicularis and the outgroup are illustrated in different colors. The studied sequences are bold, with black circles, and placed in two clades, colored red.
Fig 6.
Median-joining network analysis of haplotype inferred from cox1 sequences (333 bp) of E. vermicularis using PopArt 1.7 software (http://popart.otago.ac.nz).
Eighty-seven sequences were distributed among 59 haplotypes (12 sequences from this study and the rest from other studies in the NCBI database). All the sequences from this study belonged to type B. Each circle represents a haplotype, and its colors represent a country. The size of the circle indicates the relative frequency of sequences belonging to a particular haplotype. Hatch marks along the network branches indicate the number of mutations. B. Distribution and diversity of different haplotypes of E. vermicularis based on locations on the world map. Each color represents a different geographical location. PopArt software was used to place sample locations for sequences on the map panel (http://popart.otago.ac.nz/index.shtml) [28]. The map interface was implemented using the Marble library (http://marble.kde.org), which is distributed according to the GNU Lesser General Public License (LGPL), Version 2 (http://www.gnu.org/licenses/lgpl.html). Map data is copyright OpenStreetMap and was used under the Open Database License (www.opendatacommons.org/licenses/odbl).