2.2. Sand fly collection and species identification
Sand flies were caught in both localities using CDC light traps inside households and sticky papers outside dwellings, from June 20 to July 10 and from September 20 to October 10, 2015. In El Borouj, houses with and without ACL cases were sampled. One to two CDC traps were set in each selected house for one night under favourable weather conditions. Sticky traps consisted of 21x29.5 cm sheets of papers covered in castor oil, 9-17 were set the same day in adult sand fly resting places (holes in house walls and on other nearby walls) and left for four days. The captured sand flies were stored in 70% alcohol. Male and female specimens were separated and morphologically identified using taxonomic keys (Rioux and Golvan, 1969; Rioux et al., 1978; Leger et al., 1983; El Sawaf et al., 1989; Gil Collado et al.,1989; Benabdennbi et al., 1999; Berchi et al., 2007; Sáezet al., 2018). The specimens were placed in Marc André solution and heated to boiling point, and finally mounted on slides under a coverslip using Berlese solution. The genitalia of P. sergentispecimens was individually removed and mounted on slides under a coverslip for morphological identification whereas the rest of the body was stored at -20ºC for DNA extraction.
The gonotrophic cycle of the female sand flies was categorised as blood-fed, non-fed or gravid. Density (sand flies/trap), relative abundance (% specimens of a given species/total sand flies), and frequency (% positive sampling stations for a given species) data were estimated by species.