Table 1.
S. mansoni (Sm) and S. haematobium (Sh) like-eggs processed, according to their shedding way (urine or stool) and patient country of origin.
Fig 1.
Photographs illustrating the morphology of Schistosoma mansoni-like eggs shed by the urine of migrant patients residing in Spain.
A–C: from 3 different patients from Senegal; D: from a patient from Ivory Coast; E: from a patient from Guinea-Bissau; F: miracidium hatching under microscope light, in one egg from Senegal. (optical microscopy, original magnification, A–F = ×400; no stain used). Scale bars: A–F = 50 μm.
Table 2.
Genetic profiles obtained according to their mito-nuclear signature, and organized by migrant country of origin, egg shedding morphology and type of sample processed (urine or stool).
Table 3.
ITS-2 sequences identified in the 86 clonal sequences derived from 13 S. mansoni-like eggs.
Table 4.
Molecular haplotype identification by rDNA (ITS1-5.8S-ITS2) and mtDNA cox1 sequencing of lateral and terminal spined eggs, according to the type of sample (urine or stool), patient, and country of origin. Codes for patients listed in Table 1.
Fig 2.
Polymorphic sites in the sequence comparison of the complete transcribed spacer region of the nuclear rDNA between the haplotypes of A) S. mansoni like-eggs and B) S. haematobium like-eggs haplotypes (H) obtained (in bold) and other haplotypes or isolates from GenBank (S1 Table).
Numbers (to be read in vertical) refer to variable positions in the alignment made with MEGA X;. = Identical; - = Indel; - = Not sequenced; N = Undetermined nucleotide base; Heterozygotic position/s represented with corresponding symbol of IUPAC code for incomplete nucleic acid specification. Ci = Côte d’Ivoire; Gb = Guinea-Bissau; Ma=Mali; Se = Senegal.
Fig 3.
Two different fragments of ITS chromatograms showing the onset of the overlapping reading with the A) forward and B) reverse primers, respectively, inS. mansoni x S. haematobium sequences.
The overlapping chromatograms reading with both primers is caused by two deletions in the S. haematobium sequence or two insertions in the S. mansoni sequence at positions 80 (brown arrow) (C) and 284 (yellow arrow) (D) of their alignment. This does not allow the nucleotide assignment from positions 80 to 283 of the 5’ ITS-1 region in eggs containing an S. mansoni x S. haematobium ITSs sequence. Black arrows indicate heterozygous positions at sites that discriminate between S. mansoni and S. haematobium.
Fig 4.
Polymorphic sites in nucleotide and amino acid positions identified in the cox1 mtDNA sequence alignment of S. mansoni-like and S. haematobium-like haplotypes
(H) (in bold) and isolates of the same species from GenBank. Numbers (to be read in vertical) refer to variable positions obtained in the alignment made with MEGA X.= Identical; - = Not sequenced;? = undetermined positions. In the case of identical sequences (S2 Table), only one GenBank accession number has been selected as representative.
Fig 5.
Phylogenetic tree of mtDNA cox1 of Schistosoma species based on maximum-likelihood model constructed with 11 haplotypes (7 Schistosoma mansoni and 4 S. haematobium) from the present study (in bold) and 30 sequences from the Genbank database (accession numbers are shown in the tree).
Tree rooted using the S. japonicum sequence (KU196417) as outgroup. The tree is drawn to scale, with branch lengths measured as the number of substitutions per site. Bootstrap supports for nodes obtained using 1,000 replicates.