Fig 1.
Map of Vietnam showing tick collection sites realised with QGIS version 3.10, the layers have been uploaded to the DIVA-GIS website: https://www.diva-gis.org/datadown (A); Morphologically, the 70% alcohol tick-preserved species were collected in Vietnam over a period of 10 years: Amblyomma varanensis [♀: a, b]; Amblyomma sp. [♀: c, d]; Dermacentor auratus [♂: e, f]; Dermacentor compactus [♂: g, h]; and approximately 2 years: Rhipicephalus (B) microplus [♀: I, k]; Rhipicephalus sanguineus s.l [♂: l, m] (B).
Table 1.
Target amplified and used for qPCR and standard PCR.
Table 2.
The number of tick species used for MALDI-TOF MS analysis, creation of the MS reference spectra creation, and molecular biology confirmation.
Fig 2.
Comparison of MALDI-TOF MS spectra from the legs of six tick species collected in Vietnam.
The MS spectra revealed intra-species reproducibility and inter-species specificity (A); The MS spectra were compared by Principal Component Analysis (B); a.u., arbitrary units; m/z, mass-to-charge ratio.
Fig 3.
Comparison of MALDI-TOF MS spectra from the legs of six alcohol-preserved tick species collected in Vietnam and stored for different periods of time.
The dendrogram was built using between one and eight representative MS spectra from six distinct tick species (A). The MS spectra of different specimens showed intra-species reproducibility and inter-species specificity as confirmed by PCA (B).
Fig 4.
23S rRNA gene-based phylogenetic analysis of strains identified in this study.
Phylogenetic tree highlighting the position of A. phagocytophilum, A. marginale, A. platys, and E. rustica identified in our study are close to their homologues available in GenBank (A). 18S rRNA gene-based phylogenetic analysis of strains identified in the present study. Phylogenetic tree highlighting the position of B. vogeli, T. sinensis, and T. orientalis relative to their correspondence available in GenBank (B).
Table 3.
Microorganisms detected using molecular biology tools in ticks collected in Vietnam.