Analysing the role of semiochemicals in the oviposition substrate choices of the malaria vector mosquito Anopheles gambiae sensu lato

The search for tools that target malaria vector that resist insecticides and bite outdoors has become a research priority. Such tools will be necessary for managing residual malaria transmission and hastening the eradication of this devastating disease. This study investigated chemicals that potentially affect the oviposition substrate choices of Anopheles gambiae sensu lato (s.l.). It is foreseen that increased knowledge of the oviposition behaviour of this major malaria vectors and chemicals cues that mediate oviposition site-selection can be applied in the development of additional sampling methods and alternative interventions that to trap gravid malaria mosquitoes outdoors. To achieve a reproducible high egg-laying success of An. gambiae sensu stricto (s.s.) and An. arabiensis four factors were evaluated: (1) the time provided for mating; (2) the impact of cage size, mosquito age and female body size on insemination; (3) the peak oviposition time; and, (4) the host source of blood meals. Then four bioassays were optimised for studying oviposition responses of An. gambiae s.s. in the laboratory and semi-field conditions: a WHO-tube bioassay and a wind-tunnel that detected short-range attraction in the laboratory; a two-tier choice egg-count bioassay that compared the relative proportion of eggs laid in substrates in the laboratory; and a modified BG Sentinel mosquito gravid trap that evaluated long-range attraction of gravid females to olfactory cues in the semi-field. Finally, the oviposition responses of gravid An. gambiae s.s. mosquitoes to water vapour, Bermuda grass hay infusion (hay infusion), and putative semiochemicals identified from the hay infusion and a soil infusion previously shown to elicit higher egg deposition compared to filtered Lake Victoria water (lake water) in two choice egg-count bioassays (Herrera-Varela et al. 2014), were evaluated. High oviposition rates [84%, 95% confidence interval (CI) 77-89%] were achieved when 300 male and 300 blood-fed female An. gambiae s.s. were held together in a cage for four days. The chance of oviposition in the mosquitoes dropped when human host source of blood-meal was substituted with a rabbit (Odds ratio (OR) 0.30, 95% CI 0.14-0.66) but egg-numbers per female were not affected. All four optimised oviposition bioassays effectively showed between 15-20% shifts in oviposition substrate choices of mosquitoes with 80% statistical power and 5% significance. Using the WHO-tube bioassay, gravid An. gambiae s.s. were shown to be 2.4 times (95% CI 1.3-4.7 times) more likely to move towards high humidity in still air compared to non-gravid Preamble mosquitoes. This was more pronounced in the airflow olfactometer where the gravid mosquitoes were 10.6 times (95% CI 5.4-20.8 times) more likely to fly into a chamber with water than a dry chamber. Two-choice egg-count bioassays showed that An. gambiae s.s. were less likely to lay eggs in six-day old hay infusion (OR 0.10, 95% CI 0.03-0.33) compared to lake water. Ten putative semiochemicals were identified from the hay infusion using mass spectrometry and published electrophysiology data: 4-hepten-1-ol, 4-ethylphenol, phenylmethanol, 2-phenylethanol, indole, phenol, 3-methylindole, 3-methyl-1-butanol, 4-ethylphenol, and nonanal. Tested in two-choice egg-count bioassays, the first four listed compounds had no effect on egg deposition at the tested concentrations (between 0.01-5 parts per million) but mosquitoes were less likely to lay eggs in at least one concentration of 3-methylindole (OR 0.39, 95% CI 0.21-0.71), indole (OR 0.57, 95% CI 0.37-0.87), 3-methyl-1-butanol (OR 0.32, 95% CI 0.22-0.47), phenol (OR 0.55, 0.32-0.95), 4-methylphenol (OR 0.32, 0.18-0.57) and nonanal (OR 0.66, 95% CI 0.47-0.91) compared to lake water. In contrast to the hay infusion and hay infusion volatiles, An. gambiae s.s. were about two times more likely to lay eggs in cedrol, a sesquiterpene alcohol identified from the soil infusion, compared to lake water (OR 1.84, 95% CI 1.16-2.91). Cedrol attracted twice as many gravid mosquitoes in the semi-field also (OR 1.92, 95% CI 1.63-2.27). In the field, modified BG-Sentinel traps, electrocuting nets and OviART gravid traps with lake water and cedrol were three times more likely to trap malaria mosquitoes compared to traps with water only (OR 3.3, 95% CI 1.4-7.9). In conclusion, water vapour was shown to be a strong, non-specific pre-oviposition attractant for gravid An. gambiae s.s. in still air and moving air. It is probably the long range cue that gravid An. gambiae s.l. use to detect the presence aquatic habitats beyond the range of chemical cues. Evidence showed that An. gambiae s.s. discriminate between potential oviposition substrates and that this selective process is in-part mediated by volatile organic compounds originating from the site. Water vapour leads gravid mosquitoes to aquatic sites but semiochemicals enable the mosquitoes to discriminate and select between potential habitats. It was demonstrated that synthetic equivalents of semiochemicals found to attract gravid mosquitoes such as cedrol can be used to trap malaria mosquitoes outdoors

An assessment of opportunities and challenges for public sector involvement in the maternal health voucher program in Uganda

This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background: Continued inequities in coverage, low quality of care, and high out-of-pocket expenses for health services threaten attainment of Millennium Development Goals 4 and 5 in many sub-Saharan African countries. Existing health systems largely rely on input-based supply mechanisms that have a poor track record meeting the reproductive health needs of low-income and underserved segments of national populations. As a result, there is increased interest in and experimentation with results-based mechanisms like supply-side performance incentives to providers and demand-side vouchers that place purchasing power in the hands of low-income consumers to improve uptake of facility services and reduce the burden of out-of-pocket expenditures. This paper describes a reproductive health voucher program that contracts private facilities in Uganda and explores the policy and implementation issues associated with expansion of the program to include public sector facilities. Methods: Data presented here describes the results of interviews of six district health officers and four health facility managers purposefully selected from seven districts with the voucher program in southwestern Uganda. Interviews were transcribed and organized thematically, barriers to seeking RH care were identified, and how to address the barriers in a context where voucher coverage is incomplete as well as opportunities and challenges for expanding the program by involving public sector facilities were investigated. Results: The findings show that access to sexual and reproductive health services in southwestern Uganda is constrained by both facility and individual level factors which can be addressed by inclusion of the public facilities in the program. This will widen the geographical reach of facilities for potential clients, effectively addressing distance related barriers to access of health care services. Further, intensifying ongoing health education, continuous monitoring and evaluation, and integrating the voucher program with other services is likely to address some of the barriers. The public sector facilities were also seen as being well positioned to provide voucher services because of their countrywide reach, enhanced infrastructure, and referral networks. The voucher program also has the potential to address public sector constraints such as understaffing and supply shortages.Conclusions: Accrediting public facilities has the potential to increase voucher program coverage by reaching a wider pool of poor mothers, shortening distance to service, strengthening linkages between public and private sectors through public-private partnerships and referral systems as well as ensuring the awareness and buy-in of policy makers, which is crucial for mobilization of resources to support the sustainability of the programs. Specifically, identifying policy champions and consulting with key policy sectors is key to the successful inclusion of the public sector into the voucher program

The role of grass volatiles on oviposition site selection by Anopheles arabiensis and Anopheles coluzzii

Background: The reproductive success and population dynamics, of Anopheles malaria mosquitoes is strongly influenced by the oviposition site selection of gravid females. Mosquitoes select oviposition sites at different spatial scales, starting with selecting a habitat in which to search. This study utilizes the association of larval abundance in the field with natural breeding habitats, dominated by various types of wild grasses, as a proxy for oviposition site selection by gravid mosquitoes. Moreover, the role of olfactory cues emanating from these habitats in the attraction and oviposition stimulation of females was analysed. Methods: The density of Anopheles larvae in breeding sites associated with Echinochloa pyramidalis, Echinochloa stagnina, Typha latifolia and Cyperus papyrus, was sampled and the larvae identified to species level. Headspace volatile extracts of the grasses were collected and used to assess behavioural attraction and oviposition stimulation of gravid Anopheles arabiensis and Anopheles coluzzii mosquitoes in wind tunnel and two-choice oviposition assays, respectively. The ability of the mosquitoes to differentiate among the grass volatile extracts was tested in multi-choice tent assays. Results: Anopheles arabiensis larvae were the most abundant species found in the various grass-associated habitats. The larval densities described a hierarchical distribution, with Poaceae (Echinochloa pyramidalis and Echinochloa stagnina)-associated habitat sites demonstrating higher densities than that of Typha-associated sites, and where larvae were absent from Cyperus-associated sites. This hierarchy was maintained by gravid An. arabiensis and An. coluzzii mosquitoes in attraction, oviposition and multi-choice assays to grass volatile extracts. Conclusions: The demonstrated hierarchical preference of gravid An. coluzzii and An. arabiensis for grass volatiles indicates that vegetation cues associated with larval habitats are instrumental in the oviposition site choice of the malaria mosquitoes. Identifying volatile cues from grasses that modulate gravid malaria mosquito behaviours has distinct potential for the development of tools to be used in future monitoring and control methods