Fig 1.
Flowchart of methods and samples used, divided by sections: reptile capturing and processing, entomological survey and dog sampling.
Created in: https://biorender.com/.
Fig 2.
Geographic location of reptile and dog blood sample collection sites and sand fly capturing, in the surroundings of Valenzano municipality, Apulia, Region.
(Publicly available satellite shapefiles from http://mt0.google.com/vt/lyrs=s&hl=en&x={x}&y={y}&z={z}; in QGIS 3.4).
Fig 3.
Environmental characteristics of both collection sites.
A) “muretto a secco” where the CDC light traps were placed in the campus of Veterinary Medicine, University of Bari “Aldo Moro”. B) a high-walled private-owned dog shelter.
Fig 4.
Trypanosomatidae from geckoes.
A) Trypanosoma sp. trypomastigotes in blood of Tarentola mauritanica gecko. B) molecularly identified Leishmania infantum amastigote-like forms in leukocyte of Tarentola mauritanica gecko. Scale bars A) 50 μm; B) 10 μm.
Table 1.
Trypanosomatid detection in Tarentola mauritanica geckoes using blood cytology, isolation (in TE medium), dqPCR and qPCR; values of the threshold cycle (Ct) are reported.
Table 2.
Sand fly females tested for Leishmania infantum and/or Leishmania tarentolae by duplex quantitative PCR.
The mean (M), minimum (Min), maximum (Max) and standard deviation (sd) values of the threshold cycle (Ct) are reported.
Table 3.
Antibody titers against Leishmania infantum and Leishmania tarentolae promastigotes detected by indirect fluorescent antibody test (IFAT) according to sampling time (May and November 2021) and serum dilution (1:80 to 1:2560).
Values of the threshold cycle (Ct) are reported for dqPCR and qPCR that were only positive for Leishmania infantum.
Fig 5.
Cultured promastigotes of isolated strains.
A) the strain RTAR/IT/21/RI325 isolated from a gecko Tarentola mauritanica, with longer body and flagella. B) the strain ISER/IT/21/SF178 isolated from a sand fly female Sergentomyia minuta. Scale bars A,B) 30 μm.
Fig 6.
Phylogenetic tree based on Leishmania hsp70 sequences inferred using the maximum likelihood method based on the Kimura 3-parameter model.
Trypanosoma brucei gambiense was used as an outgroup. Scale bar indicates nucleotide substitution per site. The sequences of Leishmania spp. obtained in this study are in bold.