Most malaria vectors belong to species complexes. Sibling species often exhibit divergent behaviors dictating the measures that can be deployed effectively in their control. Despite the importance of the Anopheles funestus complex in malaria transmission in sub-Saharan Africa, sibling species have rarely been identified in the past and their vectoring potential remains understudied. We analyzed 1149 wild-caught An. funestus (senso lato) specimens from 21 sites in Kenya, covering the major malaria endemic areas including western, central and coastal areas. Indoor and outdoor collection tools were used targeting host-seeking and resting mosquitoes. The identity of sibling species, infection with malaria Plasmodium parasites, and the host blood meal sources of engorged specimens were analyzed using PCR-based and sequencing methods. The most abundant sibling species collected in all study sites were Anopheles funestus (59.8%) and Anopheles rivulorum (32.4%) among the 1062 successfully amplified specimens of the An. funestus complex. Proportionally, An. funestus dominated in indoor collections whilst An. rivulorum dominated in outdoor collections. Other species identified were Anopheles leesoni (4.6%), Anopheles parensis (2.4%), Anopheles vaneedeni (0.1%) and for the first time in Kenya, Anopheles longipalpis C (0.7%). Anopheles funestus had an overall Plasmodium infection rate of 9.7% (62/636), predominantly Plasmodium falciparum (59), with two infected with Plasmodium ovale and one with Plasmodium malariae. There was no difference in the infection rate between indoor and outdoor collections. Out of 344 An. rivulorum, only one carried P. falciparum. We also detected P. falciparum infection in two non-blood-fed An. longipalpis C (2/7) which is the first record for this species in Kenya. The mean human blood indices for An. funestus and An. rivulorum were 68% (93/136) and 64% (45/70), respectively, with feeding tendencies on a broad host range including humans and domestic animals such as cow, goat, sheep, chicken and pig. Our findings underscore the importance of active surveillance through application of molecular approaches to unravel novel parasite-vector associations possibly contributed by cryptic species with important implications for effective malaria control and elimination.

Jingmen tick virus (JMTV) is an arbovirus with a multisegmented genome related to those of unsegmented flaviviruses. The virus first described in Rhipicephalus microplus ticks collected in Jingmen city (Hubei Province, China) in 2010 is associated with febrile illness in humans. Since then, the geographic range has expanded to include Trinidad and Tobago, Brazil, and Uganda. However, the ecology of JMTV remains poorly described in Africa. We screened adult ticks (n = 4550, 718 pools) for JMTV infection by reverse transcription polymerase chain reaction (RT-PCR). Ticks were collected from cattle (n = 859, 18.88%), goats (n = 2070, 45.49%), sheep (n = 1574, 34.59%), and free-ranging tortoises (Leopard tortoise, Stigmochelys pardalis) (n = 47, 1.03%) in two Kenyan pastoralist-dominated areas (Baringo and Kajiado counties) with a history of undiagnosed febrile human illness. Surprisingly, ticks collected from goats (0.3%, 95% confidence interval (CI) 0.1–0.5), sheep (1.8%, 95% CI 1.2–2.5), and tortoise (74.5%, 95% CI 60.9–85.4, were found infected with JMTV, but ticks collected from cattle were all negative. JMTV ribonucleic acid (RNA) was also detected in blood from tortoises (66.7%, 95% CI 16.1–97.7). Intragenetic distance of JMTV sequences originating from tortoise-associated ticks was greater than that of sheep-associated ticks. Phylogenetic analyses of seven complete-coding genome sequences generated from tortoise-associated ticks formed a monophyletic clade within JMTV strains from other countries. In summary, our findings confirm the circulation of JMTV in ticks in Kenya. Further epidemiological surveys are needed to assess the potential public health impact of JMTV in Kenya.

The transmission patterns of dengue (DENV) and yellow fever (YFV) viruses, especially in urban settings, are influenced by Aedes (Stegomyia) mosquito abundance and behavior. Despite recurrent dengue outbreaks on the Kenyan coast, these parameters remain poorly defined in this and other areas of contrasting dengue endemicity in Kenya. In assessing the transmission risk of DENV/YFV in three Kenyan cities, we determined adult abundance and resting habits of potential Aedes (Stegomyia) vectors in Kilifi (dengue-outbreak prone), and Nairobi and Kisumu (no dengue outbreaks reported). In addition, mosquito diversity, an important consideration for changing mosquito-borne disease dynamics, was compared. Between October 2014 and June 2016, host-seeking adult mosquitoes were sampled using CO2-baited BG-Sentinel traps (12 traps daily) placed in vegetation around homesteads, across study sites in the three major cities. Also, indoor and outdoor resting mosquitoes were sampled using Prokopack aspirators. Three samplings, each of five consecutive days, were conducted during the long-rains, short-rains and dry season for each city. Inter-city and seasonal variation in mosquito abundance and diversity was evaluated using general linear models while mosquito-resting preference (indoors vs outdoors) was compared using Chi-square test. Aedes aegypti, which comprised 60% (n = 7772) of the total 12,937 host-seeking mosquitoes collected, had comparable numbers in Kisumu (45.2%, n = 3513) and Kilifi (37.7%, n = 2932), both being significantly higher than Nairobi (17.1%, n = 1327). Aedes aegypti abundance was significantly lower in the short-rains and dry season relative to the long-rains (P < 0.0001). Aedes bromeliae, which occurred in low numbers, did not differ significantly between seasons or cities. Mosquito diversity was highest during the long-rains and in Nairobi. Only 10% (n = 43) of the 450 houses aspirated were found positive for resting Ae. aegypti, with overall low captures in all areas. Aedes aegypti densities were comparable indoors/outdoors in Kilifi; but with higher densities outdoors than indoors in Kisumu and Nairobi. The presence and abundance of Ae. aegypti near human habitations and dwellings, especially in Kilifi/Kisumu, is suggestive of increased DENV transmission risk due to higher prospects of human vector contact. Despite low abundance of Ae. bromeliae suggestive of low YFV transmission risk, its proximity to human habitation as well as the observed diversity of potential YFV vectors should be of public health concern and monitored closely for targeted control. The largely outdoor resting behavior for Ae. aegypti provides insights for targeted adult vector control especially during emergency outbreak situations.

Sandfly-Associated Phlebovirus in Humans, Kenya

Malaria is a major public health problem in Ghana. We present a site-specific entomological study of malaria vectors and transmission indices as part of an effort to develop a site for the testing of improved control strategies including possible vaccine trials.