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Background   Phlebotomus (Larroussius) guggisbergi  is among the confirmed vectors for cutaneous leishmaniasis (CL) transmission in Kenya. This scarring and stigmatizing form of leishmaniasis accounts for over one million annual cases worldwide. Most recent CL epidemics in Kenya have been reported in Gilgil, Nakuru County, where the disease has become a public health issue. However, little is known about the factors that drive its transmission. Here, we sought to determine the occurrence, distribution and host blood feeding preference of the vectors, and to identify  Leishmania  species and infection rates in sandflies using molecular techniques. This information could lead to a better understanding of the disease transmission and improvement of control strategies in the area.    Methodology/ Principal findings  An entomological survey of sandflies using CDC light traps was conducted for one week per month in April 2016, and in June and July 2017 from five villages of Gilgil, Nakuru county; Jaica, Sogonoi, Utut, Gitare and Njeru. Sandflies were identified to species level using morphological keys and further verified by PCR analysis of  cytochrome c oxidase subunit I (COI)  gene. Midguts of female sandflies found to harbour  Leishmania  were ruptured and the isolated parasites cultured in Novy-MacNeal-Nicolle (NNN) media overlaid with Schneider’s insect media to identify the species.  Leishmania  parasite screening and identification in 198 randomly selected  Phlebotomus  females and parasite cultures was done by PCR-RFLP analysis of  ITS1  gene, nested kDNA-PCR and real-time PCR-HRM followed by sequencing. Bloodmeal source identification was done by real-time PCR-HRM of the vertebrate  cytochrome-b  gene. A total of 729 sandflies (males: n = 310; females: n = 419) were collected from Utut (36.6%), Jaica (24.3%), Sogonoi (34.4%), Njeru (4.5%), and Gitare (0.1%). These were found to consist of nine species: three  Phlebotomus  spp. and six  Sergentomyia  spp.  Ph .  guggisbergi  was the most abundant species (75.4%, n = 550) followed by  Ph .  saevus sensu lato  (11.3%, n = 82). Sandfly species distribution across the villages was found to be significantly different (p<0.001) with Jaica recording the highest diversity. The overall  Leishmania  infection rate in sandflies was estimated at 7.07% (14/198). Infection rates in  Ph .  guggisbergi  and  Ph .  saevus s . l . were 9.09% (12/132) and 3.57% (2/56) respectively.  L .  tropica  was found to be the predominant parasite in Gilgil with an overall infection rate of 6.91% (13/188) in  Ph .  guggisbergi  (n = 11) and  Ph .  saevus s . l . (n = 2) sandflies. However, PCR analysis also revealed  L .  major  infection in one  Ph .  guggisbergi  specimen. Bloodmeal analysis in the 74 blood-fed sandflies disclosed a diverse range of vertebrate hosts in  Ph .  guggisbergi  bloodmeals, while  Ph .  saevus s . l . fed mainly on humans.    Conclusions/ Significance  The high infection rates of  L .  tropica  and abundance of  Ph .  guggisbergi  in this study confirms this sandfly as a vector of  L .  tropica  in Kenya. Furthermore, isolation of live  L .  tropica  parasites from  Ph .  saevus s . l . suggest that there are at least three potential vectors of this parasite species in Gilgil;  Ph .  guggisbergi ,  Ph .  aculeatus  and  Ph .  saevus s . l . Molecular identification of  L .  major  infections in  Ph .  guggisbergi  suggested this sandfly species as a potential permissive vector of  L .  major , which needs to be investigated further. Sandfly host preference analysis revealed the possibility of zoonotic transmissions of  L .  tropica  in Gilgil since the main vector ( Ph .  guggisbergi ) does not feed exclusively on humans but also other vertebrate species. Further investigations are needed to determine the potential role of these vertebrate species in  L .  tropica  and  L .  major  transmission in the area.

plos neglected tropical diseases

Association of Phlebotomus guggisbergi with Leishmania major and Leishmania tropica in a complex transmission setting for cutaneous leishmaniasis in Gilgil, Nakuru county, Kenya