Human biting rhythm of Anopheles gambiae Giles, 1902 (Diptera: Culicidae) and sleeping behaviour of pregnant women in a lagoon area in Southern Benin

Objective In the framework of EVALMOUS study aiming to assess the use and effectiveness of mosquito nets by pregnant women and other members of their household in a lagoon area in southern Benin, the behaviour of pregnant women relative to the time they go to bed using the net were recorded. Malaria vectors biting rhythm, Plasmodium falciparum infection and insecticide resistance genes in malaria vectors were also determined. Results Overall, 3848 females of Anopheles gambiae s. l were collected and 280 pregnant women responded to the survey. Almost all Anopheles gambiae s. l. tested were Anopheles coluzzi Coetzee and Wilkerson 2013 (Diptera: Culicidae). The CSP index in malaria vector was 1.85% and the allelic frequency of kdr gene was 74.4%. Around 90% of bites and Plasmodium falciparum Welch, 1897 (Haemosporida: Plasmodiidae) transmission occurred between 10 p.m. and 6 a.m., which coincides with the period when more than 80% of pregnant women were under bednet. Despite a slight early evening and early morning biting activity of malaria vectors in the study area, the good use of nets might remain a useful protection tool against mosquito biting and malaria transmission.

Potential of the Ethanol Extract of Aerial Parts of Diplazium esculentum (Retz) SW. AS Larvicide against Anopheles gambiae Giles and Culex quinquefasciatus Say

Plant products have been tested as insecticides against mosquitoes as they are promising candidates to replace conventional insecticides. This study was carried out to evaluate the larvicidal potential of ethanol extract of the aerial parts of Diplazium esculentum against Anopheles gambiae and Culex quinquefasciatus. Ethanol extract of the aerial parts of D. esculentum was screened for its phytochemical constituents and used for larvicidal assay. A stock solution of the extract (5 g in 100 ml of water) was prepared. From the stock solution, 0.45, 0.60, 0.75, 0.90 and 1.05% w/v concentrations of the extract were obtained for the study. Each concentration of the extract had 3 replicates. The control was also replicated. Twenty (20) third instar larvae each of Anopheles gambiae and Culex quinquefasciatus were separately exposed to each extract concentration for a duration of 48 hours. Larval nutrient was added to each experimental set up. Observations were made after 24 and 48 hours exposure period. Phytochemical screening revealed the presence of some plant metabolites. Mortality of larvae exposed to the extract increased with increased concentration and exposure time. This study revealed a differential susceptibility of larvae of the two mosquito species to the extract as evident by the 24 h LC50 values obtained which were 0.355 and 2.468% w/v for An. gambiae and Cx. quinquefasciatus respectively. Exposure of An. gambiae larvae to the extract resulted in 100% mortality even with the least concentration of 0.45% w/v after 48 hours exposure period while the highest concentration of extract (1.05% w/v) resulted in 53.33% mortality of Cx. quinquefasciatus larvae, after an exposure period of 48 hours. Results obtained from this study suggest that the aerial parts of D. esculentum if further explored would be useful in the control of An. gambiae and Cx. quinquefasciatus.

Interactions of DEET and Novel Repellents With Mosquito Odorant Receptors

The carboxamide N,N-di-ethyl-meta-toluamide (DEET) is the most effective and widely used insect repellent today. However, drawbacks concerning the efficacy and the safety of the repellent have led to efforts to design new classes of insect repellents. Through quantitative structure–activity relationships, chemists have discovered two chemical groups of novel repellents: the acylpiperidines and the carboxamides, with the acylpiperidines generally more potent in biological assays. Although the exact mechanism of action of DEET and other repellents has not yet been thoroughly elucidated, previous research shows that the activity of insect odorant receptors are inhibited in the presence of repellents. The present electrophysiological study employs two-electrode voltage clamp with Xenopus laevis oocytes expressing AgOR2/AgOrco and AgOR8/AgOrco receptors to assess the effects of the novel repellents on Anopheles gambiae Giles (Insecta: Diptera: Culicidae) mosquito odorant receptors. The novel acylpiperidines and carboxamides reversibly inhibited (12–91%) odorant-evoked currents from both AgOR2/AgOrco and AgOR8/AgOrco receptors in a dose-dependent manner at all tested concentrations (30 μM to 1 mM). Furthermore, all the novel agents were more potent inhibitors of the receptors than DEET, with the acylpiperidines producing on average greater inhibition than the carboxamides. Interestingly, there was a correlation (r2 = 0.72) between the percentage inhibition of AgOR2/AgOrco receptor currents and protection times of the acylpiperidines. Our results add to existing evidence that the repellency of a compound is linked to its ability to disrupt the insect olfactory system and that the acylpiperidines could represent a class of more effective alternatives to the current gold standard, DEET.

Plasmodium falciparum (Haemosporodia: Plasmodiidae) and O’nyong-nyong Virus Development in a Transgenic Anopheles gambiae (Diptera: Culicidae) Strain

Transgenic Anopheles gambiae Giles (Diptera: Culicidae) mosquitoes have been developed that confer sexual sterility on males that carry a transgene encoding a protein which cuts ribosomal DNA. A relevant risk concern with transgenic mosquitoes is that their capacity to transmit known pathogens could be greater than the unmodified form. In this study, the ability to develop two human pathogens in these transgenic mosquitoes carrying a homing endonuclease which is expressed in the testes was compared with its nontransgenic siblings. Infections were performed with Plasmodium falciparum (Welch) and o’nyong-nyong virus (ONNV) and the results between the transgenic and nontransgenic sibling females were compared. There was no difference observed with ONNV isolate SG650 in intrathoracic infections or the 50% oral infectious dose measured at 14 d postinfection or in mean body titers. Some significant differences were observed for leg titers at the medium and highest doses for those individuals in which virus titer could be detected. No consistent difference was observed between the transgenic and nontransgenic comparator females in their ability to develop P. falciparum NF54 strain parasites. This particular transgene caused no significant effect in the ability of mosquitoes to become infected by these two pathogens in this genetic background. These results are discussed in the context of risk to human health if these transgenic individuals were present in the environment.