Ecological preferences and distribution of cave-dwelling sand flies (Diptera: Psychodidae) in pakistan

Sand flies are vectors of numerous pathogens, making them important worldwide. In Pakistan, limited studies have addressed the bionomics of cave dwelling sand flies. Our study describes the bionomics and distribution of troglophilic (cave dwelling) sand flies. A total of 2162 sand flies comprising of six species, Phlebotomus sergenti^{Parrot, 1917}, Sergentomyia hodgsoni^{(Sinton), 1933}, P. angustus^sp.n., S. dentate^{(Sinton), 1933}, P. keshishiani^{Shurenkova, 1936} and S. clydei^{(Sinton), 1928} were collected from April-October 2025 by sticky traps in 17 caves in Lower Dir, Pakistan. Five species of vertebrate animals were observed in these surveyed caves. Phlebotomus sergenti was the most abundant species, followed by S. hodgsoni. Male sand flies were more abundant (67.62%) compared to females (32.37%) and highest number of sand flies was observed in July. Peak number of sand flies was collected in porcupine and lizards caves. The highest abundance of P. sergenti was collected at 1160-1750 m altitude. Molecular sequencing of Cytc-mtDNA confirmed sand fly identification of Phlebotomus angustus within the subgenus Adlerius. The study reveals that cave-dwelling sand flies in Pakistan exhibit unique bionomics with peak activity in July, preferring caves inhabited by porcupines and lizards. Molecular characterization is also crucial for identifying sand fly species to understand their role in transmission of zoonotic diseases.