Urbanization of Aedes mosquito populations and evolution of arboviral disease risk in Africa.

The arboviral diseases dengue, chikungunya, and yellow fever are re-merging and gaining a foothold in Africa, with a significant threat of large outbreaks in urban areas. Although their emergence is intimately linked to the primary vector Aedes aegypti, which thrives in urban environments, the risk of these diseases remains substantially heterogeneous in different geographic areas. Range expansion of invasive mosquito species Aedes albopictus, and colonization of urban habitats by sylvatic and peridomestic Aedes vectors, are likely to alter the diseases' epidemiology. We discuss how a network of different vector species and perhaps vector subpopulations could interact with associated serotypes/genotypes/lineages of the causative viruses of these diseases potentially impacting transmission risk in urban landscapes with implications for disease surveillance and control.

The distinctive bionomics of Aedes aegypti populations in Africa.

Aedes aegypti is the primary vector of dengue, chikungunya, and Zika viruses of medical importance. Behavioral and biological attributes contribute to its vectorial capacity. The mosquito domestic form, which resides outside Africa (Ae. aegypti aegypti (Aaa)), is considered to breed in artificial containers in and around homes and preferentially feeds on human blood but commonly indulges in a plant diet. Potential divergence in these attributes, in sub-Saharan Africa (SSA) where Aaa coexists with the forest ecotype (Ae. aegypti formosus), should impact the vectoring ability and hence disease epidemiology. A summary of current knowledge on Ae. aegypti blood feeding, oviposition, and plant-feeding habits among SSA populations is provided in comparison with those in different geographies, globally. Emphasis is placed on improved understanding of the connection between changing subspecies adaptation in these traits and arbovirus disease risk in SSA in response to climate change and increasing urbanization, with the ultimate use of this information for effective disease control.

Risk assessment of urban yellow fever virus transmission in Kenya: is Aedes aegypti an efficient vector?

The absence of urban yellow fever epidemics in East Africa remains a mystery amidst the proliferation of Aedes aegypti in this region. To understand the transmission dynamics of the disease, we tested urban (Mombasa, Kisumu, and Nairobi) Aedes mosquito populations in Kenya for their susceptibility to an East African yellow fever virus (YFV) genotype. Overall, 22% (n = 805) of the Ae. aegypti that were orally challenged with an infectious dose of YFV had a midgut infection, with comparable rates for Mombasa and Kisumu (χ2 = 0.35, df = 1, P = 0.55), but significantly lower rates for Nairobi (χ2 ≥ 11.08, df = 1, P ≤ 0.0009). Variations in YFV susceptibility (midgut infection) among Ae. aegypti subspecies were not associated with discernable cytochrome c oxidase subunit 1 gene haplotypes. Remarkably, no YFV dissemination or transmission was observed among the orally challenged Ae. aegypti populations. Moreover, Ae. aegypti mosquitoes that were intrathoracically inoculated with YFV failed to transmit the virus via capillary feeding. In contrast, dissemination (oral exposure) and transmission (intrathoracic inoculation) of YFV was observed among a few peri-domestic Ae. bromeliae mosquitoes (n = 129) that were assessed from these urban areas. Our study highlights an inefficient urban Ae. aegypti population, and the potential for Ae. bromeliae in sustaining an urban YFV transmission in Kenya. An assessment of urban Ae. aegypti susceptibility to other YFV genotypes, and vector potential of urban Ae. bromeliae populations in Kenya is recommended to guide cost-effective vaccination.

Survival rate, blood feeding habits and sibling species composition of Aedes simpsoni complex: Implications for arbovirus transmission risk in East Africa.

Aedes simpsoni complex has a wide distribution in Africa and comprises at least three described sub-species including the yellow fever virus (YFV) vector Ae. bromeliae. To date, the distribution and relative contributions of the sub-species and/or subpopulations including bionomic characteristics in relation to YF transmission dynamics remain poorly studied. In this study conducted in two areas with divergent ecosystems: peri-urban (coastal Rabai) and rural (Rift Valley Kerio Valley) in Kenya, survival rate was estimated by parity in Ae. simpsoni s.l. mosquitoes sampled using CO2-baited BG Sentinel traps. We then applied PCR targeting the nuclear internal transcribed spacer 2 (ITS2), region followed by sequencing and phylogenetic analytics to identify the sibling species in the Ae. simpsoni complex among parous and blood fed cohorts. Our results show that Ae. bromeliae was the most dominant sub-species in both areas, exhibiting high survival rates, human blood-feeding, and potentially, high vectorial capacity for pathogen transmission. We document for the first time the presence of Ae. lilii in Kenya and potentially yet-to-be described species in the complex displaying human feeding tendencies. We also infer a wide host feeding range on rodents, reptile, and domestic livestock besides humans especially for Ae. bromeliae. This feeding trend could likely expose humans to various zoonotic pathogens. Taken together, we highlight the utility of genotype-based analyses to generate precision surveillance data of vector populations for enhanced disease risk prediction and to guide cost-effective interventions (e.g. YF vaccinations).

Invasive Alien Plants in Africa and the Potential Emergence of Mosquito-Borne Arboviral Diseases-A Review and Research Outlook.

The emergence of arthropod-borne viruses (arboviruses) as linked to land-use changes, especially the growing agricultural intensification and expansion efforts in rural parts of Africa, is of growing health concern. This places an additional burden on health systems as drugs, vaccines, and effective vector-control measures against arboviruses and their vectors remain lacking. An integrated One Health approach holds potential in the control and prevention of arboviruses. Land-use changes favour invasion by invasive alien plants (IAPs) and investigating their impact on mosquito populations may offer a new dimension to our understanding of arbovirus emergence. Of prime importance to understand is how IAPs influence mosquito life-history traits and how this may affect transmission of arboviruses to mammalian hosts, questions that we are exploring in this review. Potential effects of IAPs may be significant, including supporting the proliferation of immature and adult stages of mosquito vectors, providing additional nutrition and suitable microhabitats, and a possible interaction between ingested secondary plant metabolites and arboviruses. We conclude that aspects of vector biology are differentially affected by individual IAPs and that while some plants may have the potential to indirectly increase the risk of transmission of certain arboviruses by their direct interaction with the vectors, the reverse holds for other IAPs. In addition, we highlight priority research areas to improve our understanding of the potential health impacts of IAPs.

Entomological assessment of dengue virus transmission risk in three urban areas of Kenya.

Urbanization is one of the major drivers of dengue epidemics globally. In Kenya, an intriguing pattern of urban dengue virus epidemics has been documented in which recurrent epidemics are reported from the coastal city of Mombasa, whereas no outbreaks occur in the two major inland cities of Kisumu and Nairobi. In an attempt to understand the entomological risk factors underlying the observed urban dengue epidemic pattern in Kenya, we evaluated vector density, human feeding patterns, vector genetics, and prevailing environmental temperature to establish how these may interact with one another to shape the disease transmission pattern. We determined that (i) Nairobi and Kisumu had lower vector density and human blood indices, respectively, than Mombasa, (ii) vector competence for dengue-2 virus was comparable among Ae. aegypti populations from the three cities, with no discernible association between susceptibility and vector cytochrome c oxidase subunit 1 gene variation, and (iii) vector competence was temperature-dependent. Our study suggests that lower temperature and Ae. aegypti vector density in Nairobi may be responsible for the absence of dengue outbreaks in the capital city, whereas differences in feeding behavior, but not vector competence, temperature, or vector density, contribute in part to the observed recurrent dengue epidemics in coastal Mombasa compared to Kisumu.

Sand Fly-Associated Phlebovirus with Evidence of Neutralizing Antibodies in Humans, Kenya.

We describe a novel virus, designated Ntepes virus (NPV), isolated from sand flies in Kenya. NPV has the characteristic phlebovirus trisegmented genome architecture and is related to, but distinct from, Gabek Forest phlebovirus. Diverse cell cultures derived from wildlife, livestock, and humans were susceptible to NPV, with pronounced permissiveness in swine and rodent cells. NPV infection of newborn mice caused rapid and fatal illness. Permissiveness for NPV replication in sand fly cells, but not mosquito cells, suggests a vector-specific adaptation. Specific neutralizing antibodies were found in 13.9% (26/187) of human serum samples taken at the site of isolation of NPV as well as a disparate site in northeastern Kenya, suggesting a wide distribution. We identify a novel human-infecting arbovirus and highlight the importance of rural areas in tropical Africa for arbovirus surveillance as well as extending arbovirus surveillance to include hematophagous arthropods other than mosquitoes.

Dengue and yellow fever virus vectors: seasonal abundance, diversity and resting preferences in three Kenyan cities.

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Assessment of risk of dengue and yellow fever virus transmission in three major Kenyan cities based on Stegomyia indices.

Dengue (DEN) and yellow fever (YF) are re-emerging in East Africa, with contributing drivers to this trend being unplanned urbanization and increasingly adaptable anthropophilic Aedes (Stegomyia) vectors. Entomological risk assessment of these diseases remains scarce for much of East Africa and Kenya even in the dengue fever-prone urban coastal areas. Focusing on major cities of Kenya, we compared DEN and YF risk in Kilifi County (DEN-outbreak-prone), and Kisumu and Nairobi Counties (no documented DEN outbreaks). We surveyed water-holding containers for mosquito immature (larvae/pupae) indoors and outdoors from selected houses during the long rains, short rains and dry seasons (100 houses/season) in each County from October 2014-June 2016. House index (HI), Breteau index (BI) and Container index (CI) estimates based on Aedes (Stegomyia) immature infestations were compared by city and season. Aedes aegypti and Aedes bromeliae were the main Stegomyia species with significantly more positive houses outdoors (212) than indoors (88) (n = 900) (χ2 = 60.52, P < 0.0001). Overall, Ae. aegypti estimates of HI (17.3 vs 11.3) and BI (81.6 vs 87.7) were higher in Kilifi and Kisumu, respectively, than in Nairobi (HI, 0.3; BI,13). However, CI was highest in Kisumu (33.1), followed by Kilifi (15.1) then Nairobi (5.1). Aedes bromeliae indices were highest in Kilifi, followed by Kisumu, then Nairobi with HI (4.3, 0.3, 0); BI (21.3, 7, 0.7) and CI (3.3, 3.3, 0.3), at the respective sites. HI and BI for both species were highest in the long rains, compared to the short rains and dry seasons. We found strong positive correlations between the BI and CI, and BI and HI for Ae. aegypti, with the most productive container types being jerricans, drums, used/discarded containers and tyres. On the basis of established vector index thresholds, our findings suggest low-to-medium risk levels for urban YF and high DEN risk for Kilifi and Kisumu, whereas for Nairobi YF risk was low while DEN risk levels were low-to-medium. The study provides a baseline for future vector studies needed to further characterise the observed differential risk patterns by vector potential evaluation. Identified productive containers should be made the focus of community-based targeted vector control programs.

Vector competence of populations of Aedes aegypti from three distinct cities in Kenya for chikungunya virus.

BACKGROUND: In April, 2004, chikungunya virus (CHIKV) re-emerged in Kenya and eventually spread to the islands in the Indian Ocean basin, South-East Asia, and the Americas. The virus, which is often associated with high levels of viremia in humans, is mostly transmitted by the urban vector, Aedes aegypti. The expansion of CHIKV presents a public health challenge both locally and internationally. In this study, we investigated the ability of Ae. aegypti mosquitoes from three distinct cities in Kenya; Mombasa (outbreak prone), Kisumu, and Nairobi (no documented outbreak) to transmit CHIKV. METHODOLOGY/PRINCIPAL FINDINGS: Aedes aegypti mosquito populations were exposed to different doses of CHIKV (105.6-7.5 plaque-forming units[PFU]/ml) in an infectious blood meal. Transmission was ascertained by collecting and testing saliva samples from individual mosquitoes at 5, 7, 9, and 14 days post exposure. Infection and dissemination were estimated by testing body and legs, respectively, for individual mosquitoes at selected days post exposure. Tissue culture assays were used to determine the presence of infectious viral particles in the body, leg, and saliva samples. The number of days post exposure had no effect on infection, dissemination, or transmission rates, but these rates increased with an increase in exposure dose in all three populations. Although the rates were highest in Ae. aegypti from Mombasa at titers ≥106.9 PFU/ml, the differences observed were not statistically significant (χ2 ≤ 1.04, DF = 1, P ≥ 0.31). Overall, about 71% of the infected mosquitoes developed a disseminated infection, of which 21% successfully transmitted the virus into a capillary tube, giving an estimated transmission rate of about 10% for mosquitoes that ingested ≥106.9 PFU/ml of CHIKV. All three populations of Ae. aegypti were infectious as early as 5-7 days post exposure. On average, viral dissemination only occurred when body titers were ≥104 PFU/ml in all populations. CONCLUSIONS/SIGNIFICANCE: Populations of Ae. aegypti from Mombasa, Nairobi, and Kisumu were all competent laboratory vectors of CHIKV. Viremia of the infectious blood meal was an important factor in Ae. aegypti susceptibility and transmission of CHIKV. In addition to viremia levels, temperature and feeding behavior of Ae. aegypti may also contribute to the observed disease patterns.