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.