Fig 1.
Red stars represent the collection sites in this study. BK: Bokeo, LNT: Luangnamtha, LPB: Luangphabang, VTP: Vientiane province, VTC: Vientiane Capital, XYR: Xayaboury, and XK: Xiengkhouang. The map of the sampling sites was created using QGIS software and open-source data. The Lao administrative shapefile data was downloaded from the United Nations Office for the Coordination of Humanitarian Affairs (OCHA) website (https://data.humdata.org/dataset/cod-ab-lao), provided under free and open data (https://data.humdata.org/faqs/licenses); the topographic data of Earth’s surface was downloaded from NASA Earthdata (https://www.earthdata.nasa.gov/data/instruments/srtm) and are made available under NASA’s free and open data policy (https://www.earthdata.nasa.gov/engage/open-data-services-software/data-use-policy).
Table 1.
Main characteristics of collection sites for sandflies in Laos.
Table 2.
Number of sandfly species by sex from seven provinces.
Table 3.
Number of sandfly species by the site characteristics from seven provinces.
Fig 2.
Maximum likelihood phylogenetic tree constructed by IQ tree from 222 cyt-b sequences of 397 nt using a model that auto-selected based on Bayesian Information Criterion (BIC).
The numbers on the branches represent the bootstrap values (%) derived from 1,000 bootstrap replicates (options: -m TEST -bb 1000 -alrt 1000 -abayes). Reference sequences of each species generated in previous studies and available in GenBank were selected and included in this analysis. Green color indicates species found in this study. New species that are closely related to Se. maiae are labelled in red color. “n” indicates the number of sequences used for the analysis.
Fig 3.
Female morphological characteristics of sandflies.
Sergentomyia barraudi group (A, B): A: cibarium of Se. siamensis with anterior teeth, B: cibarium of Se. barraudi group without anterior teeth; Se. brevicaulis group (C, D): C: cibarium with one row of anterior teeth, and D: cibarium with two rows of anterior teeth; E: cibarium of Se. maiae paratype from Thailand; F: cibarium of Se. sutherlandi n. sp.; G: cibarium of Se. gemmea-like; Se. tambori (H, I): H: cibarium and I: spermathecae; and J: spermatheca of Ph. (Adlerius) sp. 1 (photo not to scale).
Fig 4.
Sergentomyia brevicaulis drawing from a Lao female.
A: head (TM7-SM-LNT-76); B: pharynx and cibarium (TM7-SM-LNT-38).; C: flagellomeres 1, 2 and 3 (= AIII, AIV and AV) (TM7-SM-LNT-80); D: palp (TM7-SM-LNT-80); E: third segment of the palp (P3) (TM7-SM-LNT-507); F: mouth parts (labrum-epipharynx, mandible, hypopharynx, maxilla, and labial furca from left to right) (TM7-SM-LNT-506); G: wing (TM7-SM-LNT-38); H: spermathecae (TM7-SM-LNT-506) and I: furca (TM7-SM-LNT-83).
Table 4.
Comparative measurements (in µm) and counts between Se. sutherlandi n. sp. and Se. maiae females.
Fig 5.
Sergentomyia sutherlandi n. sp. Female.
A: head (holotype SF22-EX651); B: pharynx and cibarium (holotype SF22-EX651) with a view showing the implantation of the pharyngeal teeth in the rectangular box; C: flagellomeres 1, 2 and 3 (= AIII, AIV and AV)(paratype EX22-EX677); D: palp (paratype EX22-EX650); E: third segment of the palp (P3)(paratype EX22-EX650); F: mouth parts (labrum-epipharynx, mandible, hypopharynx, maxilla, and labial furca from left to right)(paratype EX22-EX695); G: wing (paratype EX22-EX650); H: furca and spermathecae (paratype EX22-EX695).
Fig 6.
Maximum likelihood phylogenetic tree constructed from Trypanosoma partial ITS1 and 5.8S region.
Sequences from BLAST results were selected and included for analysis. The tree was constructed using IQ-TREE using the auto-selected model (K2P+G4) based on Bayesian Information Criterion (BIC) with parameters -m TEST -bb 1000 -alrt 1000 -abayes. The numbers on the branches represent the bootstrap values (%) derived from 1000 replicates. Sequences obtained from this study in Laos are highlighted in red. Leishmania major (FN677342) was used as the outgroup.