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Fig 1.

Experimental design for species identification of female sand flies and morphological examination and characterization of sand fly eggs.

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Fig 2.

Maximum Likelihood tree based on the Cytb sequences using the Tamura 3-parameter (T92) distance in MEGA X.

The sequences obtained in the present study are written in red.

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Table 1.

GenBank accession numbers of Cytb sequences used to identify the species of sand flies in this study.

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Fig 3.

Phlebotomus stantoni.

A. LM of head showing ciberial teeth; B. LM showing a spermatheca (arrow); C. LM showing a chorionic pattern of the egg with unconnected parallel ridges (arrowheads) and reticular pattern (arrows).

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Fig 4.

Ultrastructural morphology of Ph. stantoni eggs.

A, B. SEM of whole eggs showing chorionic sculpturing with unconnected parallel ridges and a reticular pattern on the same egg; C, D. Details of the chorionic pattern; E, F. Coarsely arranged fibrous material which formed minute protuberances between ridges; G. Anterior pole showing the aeropylar area with one aeropyle opening; H. Higher magnification of the aeropylar area delimited by a single non-columnar circular ridge; I. A reticular basal layer near the aeropylar area.

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Fig 5.

Sergentomyia khawi.

A. LM of a head showing the cibarial teeth; B. Posterior abdominal region of a gravid female sand fly with eggs inside; C. Higher magnification of the area of box I in B showing a spermatheca (arrow); D. Higher magnification of the area of box II in B showing the polygonal pattern of the sand fly egg.

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Fig 6.

Ultrastructural morphology of Se. khawi eggs.

A. SEM of a whole egg showing chorionic sculpturing with a polygonal pattern; B. Details of the general appearance of the chorionic sculpturing; C. High magnification image showing ridges formed by the intersection of single or double parallel series of rounded palisade units (arrows). Palisade units are linked or united at the top (arrowheads); D. Details of finely arranged fibrous material as a compact coat in inter-ridge areas (asterisks); E. Chorionic pattern near the pole and a thick reticular mesh conformation (arrows).

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Fig 7.

Grassomyia indica.

A. LM of head showing cibarial teeth; B. Posterior abdominal region of a gravid female sand fly with eggs inside; C. Higher magnification from the area of box in B showing spermathecae (arrows); D. Polygonal pattern of the sand fly eggs.

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Fig 8.

Ultrastructural morphology of Gr. indica eggs.

A. SEM of whole eggs showing chorionic sculpturing with a polygonal pattern; B. Larval head protruding from the anterior pole (arrow); C. Details of the general appearance of the chorionic pattern and a thick coat material at the pole (arrows); D. High magnification image showing boundaries of polygonal shapes formed by intersection of 2–5 lines of small randomly arranged tubercles. Some tubercles project from the surface of the eggshell (arrows). Details of finely arranged fibrous material as a compact coat in inter-boundary areas (asterisks).

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Table 2.

Important details of Old World sand fly eggs examined by SEM.

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Table 2 Expand