Kenya must wake up to the threat of cryptic Anopheles species and their impact on residual malaria transmission.

Malaria remains a major health problem in Kenya despite the huge efforts put in place to control it. The non-relenting malaria threat has partly been attributed to residual malaria transmission driven by vectors that cannot effectively be controlled by the two popularly applied control methods: long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS). Reports indicate that residual transmission is widely spread in areas where malaria is endemic. This could mean that the World Health Organization's vision of a world free of malaria remains a mirage as elimination and prevention of re-establishment of malaria are rendered unachievable. Amongst the major contributors to residual malaria transmission are cryptic rare species, species of mosquitoes that are morphologically indistinguishable, but isolated genetically, that have not been the focus of malaria control programs. Recent studies have reported extensive new Anopheles cryptic species believed to be involved in malaria transmission in Kenya. This underscores the need to understand these malaria vector species, their distribution and bionomics and their impact on malaria transmission. This article discusses reports of these cryptic species, their importance to malaria transmission, especially in the arid and semi-arid areas, and what can be done to mitigate the situation.

Efficient agri-food waste valorization using mealworm (Coleoptera: Tenebrionidae) into nutrient-rich biomass for food and feed.

The utilization of yellow mealworm, Tenebrio molitor (Linnaeus, Coleoptera: Tenebrionidae), for food and feed is gaining interest globally. However, its production is hindered by expensive commercial diets. This study assessed mealworm growth performance, survival, bioconversion, and nutritional composition when fed on wheat bran (WB) with different inclusion levels (25%, 50%, 75%, and 100%) of Irish potato waste (PW). Results indicated that mealworms fed on diets with 25%-75% PW had increased body length and 1-2 times higher weight gain compared to sole WB and PW diets. The survival rate was 93%-94% across all diets. Mealworms fed on WB had a feed conversion ratio of 3.26, while the efficiency of diet conversion increased with PW inclusion levels. Mealworms fed on diets with 75% PW inclusion had the highest crude fat (48%) and energy levels (598 kcal/100 g), while sole WB produced mealworms with the highest crude protein (55%). The acid detergent fiber achieved using 100% WB was 2- to 3-fold higher, but the crude fiber and neutral detergent fiber did not vary significantly. Considerable amounts of lysine (1.6-2 mg/100 g), methionine (0.5-0.7 mg/100 g), leucine (1.4-2 mg/100 g), and threonine (0.8-1 mg/100 g) were achieved in the mealworm larvae. Our findings revealed that cheap agricultural by-products could be successfully used for the mass production of mealworms, substantially contributing to reduced production costs. Further exploration of the nutrient-dense mealworm larvae for the development of novel food and feed products is crucial.

Polymer Beads Increase Field Responses to Host Attractants in the Dengue Vector Aedes aegypti.

This study investigates the efficacy of three different olfactory cues - cyclohexanone, linalool oxide (LO), and 6-methyl-5-heptan-2-one (sulcatone) - in attracting Aedes aegypti, the primary vector of dengue, using BG sentinel traps in a dengue-endemic area (urban Ukunda) in coastal Kenya. Two experiments were conducted. Experiment 1 compared solid formulations of the compounds in polymer beads against liquid formulations with hexane as the solvent. CO2-baited traps served as controls. In Experiment 2, traps were baited with each compound in the polymer beads, commercial BG-Lure, and CO2. Our results indicate that CO2-baited traps recorded the greatest Ae. aegypti captures in both Experiment 1 and 2, whereas trap captures with polymer beads and solvent-based treatments were comparable. In experiment 2, polymer bead-based treatments yielded significantly greater female captures, each recording ~ 2-fold more captures than traps baited with the BG-Lure. There was no significant difference, however, between the treatments. Female Ae. aegypti captured in CO2-baited traps were mainly unfed (91%), with fewer gravid mosquitoes (6.4%) compared to traps with test compounds (range; 12.7-21.1%). Male captures were lower in LO and BG-Lure baited traps compared to other treatments. Gravimetric analysis showed LO had a slower release rate compared to other compounds. The findings suggest that host-associated compounds loaded on polymer beads are more effective in trapping Ae. aegypti than commercial BG-Lure and reveal sex-specific differences in mosquito responses. These results have implications for mosquito surveillance and control programs, highlighting the potential for selective trapping strategies.

Sandfly Blood-Feeding Habits and Competence in Transmitting Ntepes Virus, a Recently Discovered Member of the Genus Phlebovirus.

Phleboviruses transmitted by sandflies are among emerging public health threats. A novel Phlebovirus named Ntepes virus (NTPV) was recently described and found to infect humans from a wide geographic area in Kenya. However, the entomologic risk factors of this virus such as the potential vectors and the transmission cycles remain poorly defined. This study assessed the ability of the colonized sandfly Phlebotomus duboscqi to transmit NTPV and determined the bloodmeal host sources of field-collected sandflies from the area where NTPV was found in Baringo County, Kenya. Five-day old laboratory-reared P. duboscqi were orally challenged with an infectious dose of NTPV (≈106.0 pfu/ml) and incubated for up to 15 days postinfection. Individual sandflies were dissected into abdomens, legs, and salivary glands and screened for the virus infection by cell culture. Of the 205 virus-exposed sandflies, 19.5% developed non-disseminated infections in the midgut, with no evidence of virus dissemination or transmission in legs and salivary glands, respectively. The midgut infection rates decreased with increasing extrinsic incubation period (Spearman's correlation, ρ = -0.71). Blood-fed specimens analyzed by polymerase chain reaction (PCR) and sequencing of a region of the mitochondrial 12S rRNA, revealed almost exclusive feeding on humans (98%) represented by the sandflies Sergentomyia schwetzi, S. clydei, S. antennata, S. squamipleuris, S. africana, and Phlebotomus martini. One specimen of S. clydei had fed on cattle (2%). These findings suggest P. duboscqi is an incompetent laboratory vector of NTPV. The high human-feeding rate by diverse sandfly species increases the likelihood of human exposure to pathogens associated with these sandflies. Assessment of the susceptibility of Sergentomyia species to NTPV is recommended given their high human-feeding tendency.

Outdoor malaria vector species profile in dryland ecosystems of Kenya.

Outdoor biting by anopheline mosquitoes is one of the contributors to residual malaria transmission, but the profile of vectors driving this phenomenon is not well understood. Here, we studied the bionomics and genetically characterized populations of An. gambiae and An. funestus complexes trapped outdoors in three selected dryland areas including Kerio Valley, Nguruman and Rabai in Kenya. We observed a higher abundance of Anopheles funestus group members (n = 639, 90.6%) compared to those of the An. gambiae complex (n = 66, 9.4%) with An. longipalpis C as the dominant vector species with a Plasmodium falciparum sporozoite rate (Pfsp) of 5.2% (19/362). The known malaria vectors including An. funestus s.s. (8.7%, 2/23), An. gambiae (14.3%, 2/14), An. rivulorum (14.1%, 9/64), An. arabiensis (1.9%, 1/52) occurred in low densities and displayed high Pfsp rates, which varied with the site. Additionally, six cryptic species found associated with the An. funestus group harbored Pf sporozoites (cumulative Pfsp rate = 7.2%, 13/181). We detected low frequency of resistant 119F-GSTe2 alleles in An. funestus s.s. (15.6%) and An. longipalpis C (3.1%) in Kerio Valley only. Evidence of outdoor activity, emergence of novel and divergent vectors and detection of mutations conferring metabolic resistance to pyrethroid/DDT could contribute to residual malaria transmission posing a threat to effective malaria control.

Human and plant volatiles; lures for mosquito, vectors of dengue virus and malaria.

Increased outbreaks of mosquito borne diseases like the deadly parasitic disease, malaria and arboviruses like Zika, yellow fever and dengue viruses around the world have led to increased interest in traps that could effectively be used against mosquitoes. For example, a Google search at the time of this writing, asking, 'which is the best way of trapping mosquitoes?' produced 35.5 million search results. Regardless of the interest in the subject, scientists have yet to find a definitive answer to these questions. One area that has been exploited as a potential source of efficient traps for mosquitoes is host odour baits. Since mosquitoes are attracted to their hosts through odours produced by the hosts, it's highly likely that synthetic chemical blends based on host odours could provide a solution. Most mosquito species have 2 hosts: vertebrate animals and vascular plants. Amongst the vertebrates, most diseases spread by mosquitoes are to humans. Considerable research has therefore been conducted on human odours that elicit attraction in mosquitoes, with emphasis on compounds from sweat and skin. Interest on plant volatiles is currently gathering pace because unlike human odours that only attract host seeking female mosquitoes, plant odours can attract both male and female mosquitoes of all gonotrophic stages. This review article concentrates on some of the chemical compounds in human and plant host odours that have shown a potential as attractants to mosquitoes especially Aedes aegypti and Anopheles gambiae s.l.

Differential response to plant- and human-derived odorants in field surveillance of the dengue vector, Aedes aegypti.

Linalool oxide (LO) and hexanoic acid (HA) represent plant- and human-derived odorants, respectively, previously found as attractants for the dengue vector Aedes aegypti. Here, we investigated if a blend of both compounds can improve captures of this mosquito species in field trials in two dengue endemic sites, Kilifi and Busia Counties in Kenya. Ae. aegypti captures were significantly higher in Kilifi than Busia (χ21,142 = 170.63, P < 0.0001) and varied by treatments (χ25,137 = 151.19, P = 0.002). We found that CO2-baited BG Sentinel traps combined with a blend of both odorants decreased Ae. aegypti captures about 2- to 4-fold compared to captures with the individual compounds (LO or HA) used as positive controls. This was the case for all blends of LO and HA, irrespective of the doses tested. Our findings indicate that combining plant- and human-derived odors may elicit a masking effect in trapping Ae. aegypti. These results partly corroborate previous findings for malaria mosquitoes which showed that combining lures from both host sources either decreases or increases trap catches depending on the dose. Further investigations in the usefulness of combining plant and animal odorants in mosquito trapping are therefore necessary.

An improved odor bait for monitoring populations of Aedes aegypti-vectors of dengue and chikungunya viruses in Kenya.

BACKGROUND: Effective surveillance and estimation of the biting fraction of Aedes aegypti is critical for accurate determination of the extent of virus transmission during outbreaks and inter-epidemic periods of dengue and chikungunya fever. Here, we describe the development and use of synthetic human odor baits for improved sampling of adult Ae. aegypti, in two dengue and chikungunya fevers endemic areas in Kenya; Kilifi and Busia counties. METHODS: We collected volatiles from the feet and trunks of two female and two male volunteers aged between 25 and 45 years. We used coupled gas chromatography- electroantennographic detection (GC/EAD) analysis to screen for antennally-active components from the volatiles and coupled GC-mass spectrometry (GC/MS) to identify the EAD-active components. Using randomized replicated designs, we compared the efficacies of Biogents (BG) sentinel traps baited with carbon dioxide plus either single or blends of the identified compounds against the BG sentinel trap baited with carbon dioxide plus the BG commercial lure in trapping Ae. aegypti. The daily mosquito counts in the different traps were subjected to negative binomial regression following the generalized linear models procedures. RESULTS: A total of ten major EAD-active components identified by GC/MS as mainly aldehydes and carboxylic acids, were consistently isolated from the human feet and trunk volatiles from at least two volunteers. Field assays with synthetic chemicals of the shared EAD-active components identified from the feet and trunk gave varying results. Ae. aegypti were more attracted to carbon dioxide baited BG sentinel traps combined with blends of aldehydes than to similar traps combined with blends of carboxylic acids. When we assessed the efficacy of hexanoic acid detected in odors of the BG commercial lure and volunteers plus carbon dioxide, trap captures of Ae. aegypti doubled over the trap baited with the commercial BG lure. However, dispensing aldehydes and carboxylic acids together in blends, reduced trap captures of Ae. aegypti by ~45%-50%. CONCLUSIONS: Our results provide evidence for roles of carboxylic acids and aldehydes in Ae. aegypti host attraction and also show that of the carboxylic acids, hexanoic acid is a more effective lure for the vector than the BG commercial lure.

Field evaluation of natural human odours and the biogent-synthetic lure in trapping Aedes aegypti, vector of dengue and chikungunya viruses in Kenya.

BACKGROUND: Methods currently used in sampling adult Aedes aegypti, the main vector of dengue and chikungunya viruses are limited for effective surveillance of the vector and accurate determination of the extent of virus transmission during outbreaks and inter - epidemic periods. Here, we document the use of natural human skin odours in baited traps to improve sampling of adult Ae. aegypti in two different endemic areas of chikungunya and dengue in Kenya - Kilifi and Busia Counties. The chemistry of the volatiles released from human odours and the Biogent (BG)-commercial lure were also compared. METHODS: Cotton socks and T-shirts were used to obtain natural human skin volatiles from the feet and trunk of three volunteers (volunteers 1 and 2 in Kilifi and volunteers 2 and 3 in Busia). Using Latin square design, we compared the efficacies of BG sentinel traps baited with carbon dioxide plus (a) no bait, (b) human feet volatiles, (c) human trunk volatiles each against (c) a control (Biogent commercial lure) at the two sites. Coupled gas chromatography-mass spectrometry (GC-MS) was used to identify and compare candidate attractants released by the commercial lure and human odours. RESULTS: Ae. aegypti captured in the trap baited with feet odours from volunteer 2 and trunk odours from the same volunteer were significantly higher than in the control trap in Busia and Kilifi respectively, [IRR = 5.63, 95% CI: 1.15 - 28.30, p = 0.030] and [IRR = 3.99, 95% CI: 0.95-16.69, p = 0.049]. At both sites, Ae. aegypti captures in traps baited with either the feet or trunk odours from volunteers 1 and 3 were not significantly different from the control. Major qualitative differences were observed between the chemical profiles of human odours and the commercial BG-lure. Aldehydes, fatty acids and ketones dominated human odour profiles, whereas the BG-lure released mainly hexanoic acid. CONCLUSIONS: Our results suggest that additional candidate attractants are present in human skin volatiles which can help to improve the efficacy of lures for trapping and surveillance of Ae. aegypti.

The role of cow urine in the oviposition site preference of culicine and Anopheles mosquitoes.

BACKGROUND: Chemical and behavioural ecology of mosquitoes plays an important role in the development of chemical cue based vector control. To date, studies available have focused on evaluating mosquito attractants and repellents of synthetic and human origins. This study, however, was aimed at seasonal evaluation of the efficiency of cow urine in producing oviposition cues to Anopheles gambiae s.l. and Culex quinquefasciatus in both laboratory and field conditions. METHODS: Oviposition response evaluation in laboratory conditions was carried out in mosquito rearing cages. The oviposition substrates were located in parallel or in diagonal positions inside the cage. Urine evaluation against gravid females of An. arabiensis and Cx. quinquefasciatus was carried out at Day 1, Day 3 and Day 7. Five millilitres (mls) of cow urine was added to oviposition substrate while de-chlorinated water was used as a control. In field experiments, 500 mls of cow urine was added in artificial habitats with 2500 mls of de-chlorinated water and 2 kgs of soil. The experiment was monitored for thirty consecutive days, eggs were collected daily from the habitats at 7.00 hrs. Data analysis was performed using parametric and non-parametric tests for treatments and controls while attraction of the oviposition substrate in each species was presented using Oviposition Activity Index (OAI). RESULTS: The OAI was positive with ageing of cattle urine in culicine species in both laboratory and field experiments. The OAI for anopheline species was positive with fresh urine. The OAI during the rainy season was positive for all species tested while in the dry season the OAI for culicine spp and Anopheles gambiae s.l., changed with time from positive to negative values.Based on linear model analysis, seasons and treatments had a significant effect on the number of eggs laid in habitats, even though the number of days had no effect. CONCLUSION: Oviposition substrates treated with cow urine in both laboratory and field conditions have shown that cow urine left to age from 1-7 days has an influence on oviposition behavioural response in mosquitoes. The analysis of microbial colonies for decaying urine should be investigated along with its associated by-products.

Trapping mosquitoes using milk products as odour baits in western Kenya.

BACKGROUND: Ample evidence has shown that blood seeking mosquitoes locate their hosts by following odours produced by the hosts. Odour baited traps would therefore, provide a solution in controlling diseases spread by mosquitoes. Comparative studies were undertaken to determine the relative efficacies of two odour baits i.e. Limburger cheese and African traditional milk cream in trapping mosquitoes in the field in western Kenya. METHOD: Comparative efficacy studies were carried out in the field using Latin square experimental designs. In the first study, a counterflow geometry (CFG) trap (MM-x model; American Biophysics Corp., USA.) baited with Limburger cheese, man landing catches (MLC), Centres for Disease Control (CDC) light trap and an entry trap were compared. In the second study, three CFG traps baited with either Limburger cheese, African traditional milk cream or with no bait were compared and in the third study four CDC traps baited with either Limburger cheese, African traditional milk cream, light or with no bait were compared. Parameters like species, catch size, abdominal status, parity status and size of the collected mosquitoes were compared. RESULTS: A total of 1,806 mosquitoes were collected (60% An. gambiae s.l and 25% An.funestus, culicines 15%). There was no significant difference in the number of An. funestus trapped by the CFG trap baited with Limburger cheese from those trapped by the MLC (P = 0.351). The Limburger cheese baited CFG trap collected significantly more gravid An. funestus than the MLC (P = 0.022). Furthermore, when the CFG trap baited with Limburger cheese and the CFG trap baited with milk cream were compared, there was no significant difference in the number of An. funestus collected (P = 0.573). The same trend was observed in the comparison of Limburger cheese baited CDC trap and milk cream baited CDC trap. CONCLUSIONS: Limburger cheese and African traditional milk cream have a potential as effective odour baits for sampling/surveillance and as oviposition attractants for the malaria vector, Anopheles funestus.