Mosquito Reproduction Research Articles

Household perceptions and subjective valuations of indoor residual spraying programmes to control malaria in northern Uganda.

BackgroundInsecticide-based tools remain critical for controlling vector-borne diseases in Uganda. Securing public support from targeted populations for such tools is an important component in sustaining their long-run effectiveness. Yet little quantitative evidence is available on the perceived benefits and costs of vector control programmes among targeted households.MethodsA survey was administered to a clustered random sample of 612 households in Gulu and Oyam districts of northern Uganda during a period of very high malaria transmission and following a pilot indoor residual spray (IRS) programme. A discrete choice experiment was conducted within the survey, in which respondents indicated their preferences for different IRS programmes relative to money compensation in a series of experimentally controlled, hypothetical choice sets. The data were analysed using conditional logit regression models to estimate respondents’ willingness to accept (WTA) some amount of money compensation in lieu of foregone malaria risk reductions. Latent class models were used to analyse whether respondent characteristics predicted WTA.ResultsAverage WTA is estimated at $8.94 annually for a 10 % reduction in malaria risk, and additional co-benefits of IRS were estimated to be worth on average $54–$56 (depending on insecticide type) per round of IRS. Significant heterogeneity is observed: Four in five household heads in northern Uganda have high valuations for IRS programmes, while the remaining 20 % experience costly side effects of IRS (valued at between $2 and $3 per round). Statistically significant predictors of belonging to the high-value group include respondent gender, mean age of household members, participation in previous IRS, basic knowledge of mosquito reproduction, and the number of mosquito nets owned. Proxies for household income and wealth are not found to be statistically significant predictors of WTA.ConclusionsThis study suggests that the majority of people in areas of high malaria transmission like northern Uganda place a high value on vector control programmes using IRS. However, there is significant heterogeneity in terms of the perceived side effects (positive and negative). This has implications for sustaining public support for these programmes in the long-term.Electronic supplementary materialThe online version of this article (doi:10.1186/s40249-016-0190-1) contains supplementary material, which is available to authorized users.

MicroRNA-309 targets the Homeobox gene SIX4 and controls ovarian development in the mosquito Aedes aegypti

Obligatory blood-triggered reproductive strategy is an evolutionary adaptation of mosquitoes for rapid egg development. It contributes to the vectorial capacity of these insects. Therefore, understanding the molecular mechanisms underlying reproductive processes is of particular importance. Here, we report that microRNA-309 (miR-309) plays a critical role in mosquito reproduction. A spatiotemporal expression profile of miR-309 displayed its blood feeding-dependent onset and ovary-specific manifestation in female Aedes aegypti mosquitoes. Antagomir silencing of miR-309 impaired ovarian development and resulted in nonsynchronized follicle growth. Furthermore, the genetic disruption of miR-309 by CRISPR/Cas9 system led to the developmental failure of primary follicle formation. Examination of genomic responses to miR-309 depletion revealed that several pathways associated with ovarian development are down-regulated. Comparative analysis of genes obtained from the high-throughput RNA sequencing of ovarian tissue from the miR-309 antagomir-silenced mosquitoes with those from the in silico computation target prediction identified that the gene-encoding SIX homeobox 4 protein (SIX4) is a putative target of miR-309. Reporter assay and RNA immunoprecipitation confirmed that SIX4 is a direct target of miR-309. RNA interference of SIX4 was able to rescue phenotypic manifestations caused by miR-309 depletion. Thus, miR-309 plays a critical role in mosquito reproduction by targeting SIX4 in the ovary and serves as a regulatory switch permitting a stage-specific degradation of the ovarian SIX4 mRNA. In turn, this microRNA (miRNA)-targeted degradation is required for appropriate initiation of a blood feeding-triggered phase of ovarian development, highlighting involvement of this miRNA in mosquito reproduction.

Dengue infection severity score - improvised disease management.

Dear editor We would like to add our views regarding the paper “Validation of Dengue infection severity score” by Pongpan et al.1 As the paper outlines, the purpose of the Dengue Severity Score is to classify individuals with dengue infection into three levels of severity with clinically acceptable underestimation or overestimation. Dengue virus is transmitted by Aedes mosquitoes, which are sensitive to various environmental conditions including temperature, rainfall, and humidity. These factors are critical to mosquito reproduction and survival, for example, higher temperatures decrease the time required for virus replication and dissemination in the Aedes mosquito.2 The incidence of dengue doubles every decade, it increased from 8.3 million apparent cases in 1990 to 58.4 million apparent cases in 2013.3 Although it is a preventable disease, dengue still has an estimated mortality rate of 20,000 people per year.2 The clinical presentation of dengue infection is characterized by fever, headache, vomiting, muscle and joint pain, as well as rash on the skin. Complicated cases may result in hemorrhagic fever, thrombocytopenia, shock, and death. The World Health Organization classification of dengue fever (2009) includes two separate entities, namely, dengue and severe dengue; however, this has yet to be proven as clinically significant. Very few studies are available, and of them, most are at a secondary or tertiary level of care and not at the level of primary care. Most studies are not carried out in resource-poor settings.4 This World Health Organization gross classification is not practical since dengue epidemic affects a wide range of age groups and manifests as different clinical presentations globally, thus adding diversity to disease spectrum.5 Dengue infection severity score is a handy assessment tool; prerequisite information required for this include patients’ clinical data consisting of their medical history, recorded vitals, complete blood count, history of bleeding episodes with results of tourniquet tests, and recent incidents of plasma leakage determined by assessing hemoconcentration, which shows a 20% increase in hematocrit. In our opinion, utilizing the clinical data in the scoring system would allow for more efficient and cost-effective patient care and may minimize unnecessary hospitalization as patients with a low score are suspected to have dengue fever and could be examined and treated as outpatients, while those with a greater score are suspected of dengue hemorrhagic fever and could be admitted in the hospital. Finally, those with the greatest score, likely to have dengue shock syndrome, could be admitted for immediate monitoring in an intensive care unit. We suggest that the scoring system should be made widely applicable to effectively manage the ongoing changes being currently observed in the virulence factors of dengue virus, for example, a fifth serotype DENV-5 recently discovered in 2015, has been shown to have caused frequent epidemics and life-threatening mortalities.5 We recommend this resourceful and impactful scoring system for efficient hospital care and management of dengue patients.

Calcium influx enhances neuropeptide activation of ecdysteroid hormone production by mosquito ovaries

A critical step in mosquito reproduction is the ingestion of a blood meal from a vertebrate host. In mosquitoes like Aedes aegypti, blood feeding stimulates the release of ovary ecdysteroidogenic hormone (OEH) and insulin-like peptide 3 (ILP3). This induces the ovaries to produce ecdysteroid hormone (ECD), which then drives egg maturation. In many immature insects, prothoracicotropic hormone (PTTH) stimulates the prothoracic glands to produce ECD that directs molting and metamorphosis. The receptors for OEH, ILP3 and PTTH are different receptor tyrosine kinases with OEH and ILP3 signaling converging downstream in the insulin pathway and PTTH activating the mitogen-activated protein kinase pathway. Calcium (Ca2+) flux and cAMP have also been implicated in PTTH signaling, but the role of Ca2+ in OEH, ILP3, and cAMP signaling in ovaries is unknown. Here, we assessed whether Ca2+ flux affects OEH, ILP3, and cAMP activity in A. aegypti ovaries and also asked whether PTTH stimulated ovaries to produce ECD. Results indicated that Ca2+ flux enhanced but was not essential for OEH or ILP3 activity, whereas cAMP signaling was dependent on Ca2+ flux. Recombinant PTTH from Bombyx mori fully activated ECD production by B. mori PTGs, but exhibited no activity toward A. aegypti ovaries. Recombinant PTTH from A. aegypti also failed to stimulate either B. mori PTGs or A. aegypti ovaries to produce ECD. We discuss the implications of these results in the context of mosquito reproduction and ECD biosynthesis by insects generally.

Knowledge Attitude and Practices Related on Prevention of Mosquito Borne Diseases and Sanitation Conditions in a Salvadoran Urban Community

Introduction: In El Salvador Mosquito Borne Diseases (MBDs) is a serious public health problem due to produce morbidity and mortality. They produce a high economic loss, which contributes to the collapse of the public health system. Over the years, the Salvadoran public health system has invested large amounts of resources to minimize the problem through campaigns against MBDs. Despite this, the population is still affected by diseases such as dengue, and more recently chikungunya and zika outbreaks. The aim of this study is to evaluate sanitation conditions variables and identify knowledge, attitudes, practices related to the prevention of MBDs. Methods: A cross-sectional survey about sanitation conditions and knowledge, attitudes and practices on prevention of MBDs of inhabitant’s ≥ 15 years from an urban community of a municipality with the highest number of cases reported in 2015. Results: Majority of respondents (98.2%) had direct supply of drinking water; however 96.4% of respondents were agreed that they had an inconsistent water supply. Regarding MBDs prevention respondents showed high acceptable knowledge (76.8%), high favorable attitude (92.1%) and an acceptable implementation of practices toward prevention reproduction of mosquitoes (58.5%) and a poor implementation of practices to prevent mosquito bites (38.3%). Conclusions: The findings revealed high acceptable knowledge about MBDs and a high favorable attitude regarding to prevent them, but also revealed a scarce implementation of prevention practices. The generalized storage of water on non-covered barrels because the inconsistent water supplies, can be source of proliferation of mosquitoes and therefore increases the risk of transmission of MBDs. High population density of the community can magnify this risk. It is necessary to enhance education to promote better implementation of practices using the most common media together with the local health workers.

SiRNA-Mediated Silencing of doublesex during Female Development of the Dengue Vector Mosquito Aedes aegypti.

The development of sex-specific traits, including the female-specific ability to bite humans and vector disease, is critical for vector mosquito reproduction and pathogen transmission. Doublesex (Dsx), a terminal transcription factor in the sex determination pathway, is known to regulate sex-specific gene expression during development of the dengue fever vector mosquito Aedes aegypti. Here, the effects of developmental siRNA-mediated dsx silencing were assessed in adult females. Targeting of dsx during A. aegypti development resulted in decreased female wing size, a correlate for body size, which is typically larger in females. siRNA-mediated targeting of dsx also resulted in decreased length of the adult female proboscis. Although dsx silencing did not impact female membrane blood feeding or mating behavior in the laboratory, decreased fecundity and fertility correlated with decreased ovary length, ovariole length, and ovariole number in dsx knockdown females. Dsx silencing also resulted in disruption of olfactory system development, as evidenced by reduced length of the female antenna and maxillary palp and the sensilla present on these structures, as well as disrupted odorant receptor expression. Female lifespan, a critical component of the ability of A. aegypti to transmit pathogens, was also significantly reduced in adult females following developmental targeting of dsx. The results of this investigation demonstrate that silencing of dsx during A. aegypti development disrupts multiple sex-specific morphological, physiological, and behavioral traits of adult females, a number of which are directly or indirectly linked to mosquito reproduction and pathogen transmission. Moreover, the olfactory phenotypes observed connect Dsx to development of the olfactory system, suggesting that A. aegypti will be an excellent system in which to further assess the developmental genetics of sex-specific chemosensation.

Substrate specificity and transport mechanism of amino-acid transceptor Slimfast from Aedes aegypti.

Anautogenous mosquitoes depend on vertebrate blood as nutrient source for their eggs. A highly efficient set of membrane transporters mediates the massive movement of nutrient amino acids between mosquito tissues after a blood meal. Here we report the characterization of the amino-acid transporter Slimfast (Slif) from the yellow-fever mosquito Aedes aegypti using codon-optimized heterologous expression. Slif is a well-known component of the target-of-rapamycin signalling pathway and fat body nutrient sensor, but its substrate specificity and transport mechanism were unknown. We found that Slif transports essential cationic and neutral amino acids with preference for arginine. It has an unusual dual-affinity mechanism with only the high affinity being Na+ dependent. Tissue-specific expression and blood meal-dependent regulation of Slif are consistent with conveyance of essential amino acids from gut to fat body. Slif represents a novel transport system and type of transceptor for sensing and transporting essential amino acids during mosquito reproduction.

Regulation of Gene Expression Patterns in Mosquito Reproduction.

In multicellular organisms, development, growth and reproduction require coordinated expression of numerous functional and regulatory genes. Insects, in addition to being the most speciose animal group with enormous biological and economical significance, represent outstanding model organisms for studying regulation of synchronized gene expression due to their rapid development and reproduction. Disease-transmitting female mosquitoes have adapted uniquely for ingestion and utilization of the huge blood meal required for swift reproductive events to complete egg development within a 72-h period. We investigated the network of regulatory factors mediating sequential gene expression in the fat body, a multifunctional organ analogous to the vertebrate liver and adipose tissue, of the female Aedes aegypti mosquito. Transcriptomic and bioinformatics analyses revealed that ~7500 transcripts are differentially expressed in four sequential waves during the 72-h reproductive period. A combination of RNA-interference gene-silencing and in-vitro organ culture identified the major regulators for each of these waves. Amino acids (AAs) regulate the first wave of gene activation between 3 h and 12 h post-blood meal (PBM). During the second wave, between 12 h and 36 h, most genes are highly upregulated by a synergistic action of AAs, 20-hydroxyecdysone (20E) and the Ecdysone-Receptor (EcR). Between 36 h and 48 h, the third wave of gene activation—regulated mainly by HR3—occurs. Juvenile Hormone (JH) and its receptor Methoprene-Tolerant (Met) are major regulators for the final wave between 48 h and 72 h. Each of these key regulators also has repressive effects on one or more gene sets. Our study provides a better understanding of the complexity of the regulatory mechanisms related to temporal coordination of gene expression during reproduction. We have detected the novel function of 20E/EcR responsible for transcriptional repression. This study also reveals the previously unidentified large-scale effects of HR3 and JH/Met on transcriptional regulation during the termination of vitellogenesis and remodeling of the fat body.

20-hydroxyecdysone stimulation of juvenile hormone biosynthesis by the mosquito corpora allata

Juvenile hormone III (JH) is synthesized by the corpora allata (CA) and plays a key role in mosquito development and reproduction. JH titer decreases in the last instar larvae allowing pupation and metamorphosis to progress. As the anti-metamorphic role of JH comes to an end, the CA of the late pupa (or pharate adult) becomes again “competent” to synthesize JH, which plays an essential role orchestrating reproductive maturation. 20-hydroxyecdysone (20E) prepares the pupae for ecdysis, and would be an ideal candidate to direct a developmental program in the CA of the pharate adult mosquito. In this study, we provide evidence that 20E acts as an age-linked hormonal signal, directing CA activation in the mosquito pupae. Stimulation of the inactive brain-corpora allata-corpora cardiaca complex (Br-CA-CC) of the early pupa (24 h before adult eclosion or −24 h) in vitro with 20E resulted in a remarkable increase in JH biosynthesis, as well as increase in the activity of juvenile hormone acid methyltransferase (JHAMT). Addition of methyl farnesoate but not farnesoic acid also stimulated JH synthesis by the Br-CA-CC of the −24 h pupae, proving that epoxidase activity is present, but not JHAMT activity. Separation of the CA-CC complex from the brain (denervation) in the −24 h pupae also activated JH synthesis. Our results suggest that an increase in 20E titer might override an inhibitory effect of the brain on JH synthesis, phenocopying denervation. All together these findings provide compelling evidence that 20E acts as a developmental signal that ensures proper reactivation of JH synthesis in the mosquito pupae.

Temporal Coordination of Carbohydrate Metabolism during Mosquito Reproduction.

Hematophagous mosquitoes serve as vectors of multiple devastating human diseases, and many unique physiological features contribute to the incredible evolutionary success of these insects. These functions place high-energy demands on a reproducing female mosquito, and carbohydrate metabolism (CM) must be synchronized with these needs. Functional analysis of metabolic gene profiling showed that major CM pathways, including glycolysis, glycogen and sugar metabolism, and citrate cycle, are dramatically repressed at post eclosion (PE) stage in mosquito fat body followed by a sharply increase at post-blood meal (PBM) stage, which were also verified by Real-time RT-PCR. Consistent to the change of transcript and protein level of CM genes, the level of glycogen, glucose and trehalose and other secondary metabolites are also periodically accumulated and degraded during the reproductive cycle respectively. Levels of triacylglycerols (TAG), which represent another important energy storage form in the mosquito fat body, followed a similar tendency. On the other hand, ATP, which is generated by catabolism of these secondary metabolites, showed an opposite trend. Additionally, we used RNA interference studies for the juvenile hormone and ecdysone receptors, Met and EcR, coupled with transcriptomics and metabolomics analyses to show that these hormone receptors function as major regulatory switches coordinating CM with the differing energy requirements of the female mosquito throughout its reproductive cycle. Our study demonstrates how, by metabolic reprogramming, a multicellular organism adapts to drastic and rapid functional changes.

Evidence based community mobilization for dengue prevention in Nicaragua and Mexico (Camino Verde, the Green Way): cluster randomized controlled trial

Objective To test whether community mobilization adds effectiveness to conventional dengue control.Design Pragmatic open label parallel group cluster randomized controlled trial. Those assessing the outcomes and analyzing the data were...

Developmental neurogenetics of sexual dimorphism in Aedes aegypti.

Sexual dimorphism, a poorly understood but crucial aspect of vector mosquito biology, encompasses sex-specific physical, physiological, and behavioral traits related to mosquito reproduction. The study of mosquito sexual dimorphism has largely focused on analysis of the differences between adult female and male mosquitoes, particularly with respect to sex-specific behaviors related to disease transmission. However, sexually dimorphic behaviors are the products of differential gene expression that initiates during development and therefore must also be studied during development. Recent technical advancements are facilitating functional genetic studies in the dengue vector Aedes aegypti, an emerging model for mosquito development. These methodologies, many of which could be extended to other non-model insect species, are facilitating analysis of the development of sexual dimorphism in neural tissues, particularly the olfactory system. These studies are providing insight into the neurodevelopmental genetic basis for sexual dimorphism in vector mosquitoes.

Frizzled 2 is a key component in the regulation of TOR signaling-mediated egg production in the mosquito Aedes aegypti

The Wnt signaling pathway was first discovered as a key event in embryonic development and cell polarity in Drosophila. Recently, several reports have shown that Wnt stimulates translation and cell growth by activating the mTOR pathway in mammals. Previous studies have demonstrated that the Target of Rapamycin (TOR) pathway plays an important role in mosquito vitellogenesis. However, the interactions between these two pathways are poorly understood in the mosquito. In this study, we hypothesized that factors from the TOR and Wnt signaling pathways interacted synergistically in mosquito vitellogenesis. Our results showed that silencing Aedes aegypti Frizzled 2 (AaFz2), a transmembrane receptor of the Wnt signaling pathway, decreased the fecundity of mosquitoes. We showed that AaFz2 was highly expressed at the transcriptional and translational levels in the female mosquito 6 h after a blood meal, indicating amino acid-stimulated expression of AaFz2. Notably, the phosphorylation of S6K, a downstream target of the TOR pathway, and the expression of vitellogenin were inhibited in the absence of AaFz2. A direct link was found in this study between Wnt and TOR signaling in the regulation of mosquito reproduction.

English

Because of climate change and failure of the existing methods of control of vector borne diseases and vector are increasing. Mosquito species are the main vectors of human pathogens causing malaria, dengue, filariasis, chikungunya, yellow fever and West Nile. There are no well-organized methods and tools of controls of vector and vector borne diseases, since no efficient vaccines or drugs are available. Despite years of intense effort to control them, many of these diseases are increasing in prevalence, geographical distribution and severity, and options to control them are limited. Currently, efforts focused on the control of vector populations. During recent years, the endosymbiont bacterium has been well-documented and has led to suggestions that these could be used to control pests and therefore diseases. Wolbachia is perhaps the most renowned insect symbiont, primarily due to its ability to manipulate insect reproduction and to interfere with major human pathogens therefore providing new avenues for pest control. Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These alphaproteobacteria endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm–egg incompatibility. Wolbachia strains can invade and sustain themselves in mosquito populations, reduce adult lifespan, affect mosquito reproduction and interfere with pathogen replication. Wolbachia can also provide direct fitness benefits to their hosts by affecting nutrition and development, influencing fecundity or oogenesis and providing resist­ance to pathogens. For instance, infection of Anopheles gambiae with both wMelPop and wAlbB reduced the oocyst burden of Plasmodium falciparum, compared to uninfected control mosquitoes. In addition, similar study observed that the wMelPop strain inhibited development of Plasmodium berghei; however, the wAlbB strain was found to enhance development of P. berghei.   Key words: Malaria, Aedes aegypti, Chikungunya, dengue, drosophila, wolbachiapipientis, vectorborn diseases.

Maternal germline-specific genes in the Asian malaria mosquito Anopheles stephensi: characterization and application for disease control.

Anopheles stephensi is a principal vector of urban malaria on the Indian subcontinent and an emerging model for molecular and genetic studies of mosquito biology. To enhance our understanding of female mosquito reproduction, and to develop new tools for basic research and for genetic strategies to control mosquito-borne infectious diseases, we identified 79 genes that displayed previtellogenic germline-specific expression based on RNA-Seq data generated from 11 life stage–specific and sex-specific samples. Analysis of this gene set provided insights into the biology and evolution of female reproduction. Promoters from two of these candidates, vitellogenin receptor and nanos, were used in independent transgenic cassettes for the expression of artificial microRNAs against suspected mosquito maternal-effect genes, discontinuous actin hexagon and myd88. We show these promoters have early germline-specific expression and demonstrate 73% and 42% knockdown of myd88 and discontinuous actin hexagon mRNA in ovaries 48 hr after blood meal, respectively. Additionally, we demonstrate maternal-specific delivery of mRNA and protein to progeny embryos. We discuss the application of this system of maternal delivery of mRNA/miRNA/protein in research on mosquito reproduction and embryonic development, and for the development of a gene drive system based on maternal-effect dominant embryonic arrest.

Three Cases of West Nile Encephalitis over an Eight-Day Period at a Downtown Los Angeles Community Hospital.

Since its introduction in New York City in 1999, the virus has spread throughout the entire North American continent and continues to spread into Central and Latin America. Our report discusses the signs and symptoms, diagnostics, and treatment of West Nile disease. It is important to recognize the disease quickly and initiate appropriate treatment. We present three cases of West Nile encephalitis at White Memorial Medical Center in East Los Angeles that occurred over the span of eight days. All three patients live within four to six miles from the hospital and do not live or work in an environment favorable to mosquitoes including shallow bodies of standing water, abandoned tires, or mud ruts. All the patients were Hispanic. Physicians and other health care providers should consider West Nile infection in the differential diagnosis of causes of aseptic meningitis and encephalitis, obtain appropriate laboratory studies, and promptly report cases to public health authorities. State governments should establish abatement programs that will eliminate sources that allow for mosquito reproduction and harboring. The public needs to be given resources that educate them on what entails the disease caused by the West Nile virus, what the symptoms are, and, most importantly, what they can do to prevent themselves from becoming infected.

Coordinated changes in JH biosynthesis and JH hemolymph titers in Aedes aegypti mosquitoes

Juvenile hormone III (JH) is synthesized by the corpora allata (CA) and plays a key role in mosquito development and reproduction. A decrease in JH titer during the last instar larvae allows pupation and metamorphosis to proceed. As the anti-metamorphic role of JH comes to an end, the CA of the late pupa once again synthesizes JH, which plays an essential role in orchestrating reproductive maturation. In spite of the importance of Aedes aegypti as a vector, a detailed study of the changes of JH hemolymph titers during the gonotrophic cycle has never been performed. In the present studies, using a high performance liquid chromatography coupled to a fluorescent detector (HPLC–FD) method, we measured changes in JH levels in the hemolymph of female mosquitoes during the pupal and adult stages. Our results revealed tightly concomitant changes in JH biosynthesis and JH hemolymph titers during the gonotrophic cycle of female mosquito. Feeding high sugar diets resulted in an increase of JH titers, and mating also modified JH titers in hemolymph. In addition these studies confirmed that JH titer in mosquitoes is fundamentally determined by the rate of biosynthesis in the CA.

Persistent oscillations and backward bifurcation in a malaria model with varying human and mosquito populations: implications for control.

We derive and study a deterministic compartmental model for malaria transmission with varying human and mosquito populations. Our model considers disease-related deaths, asymptomatic immune humans who are also infectious, as well as mosquito demography, reproduction and feeding habits. Analysis of the model reveals the existence of a backward bifurcation and persistent limit cycles whose period and size is determined by two threshold parameters: the vectorial basic reproduction number Rm, and the disease basic reproduction number R0, whose size can be reduced by reducing Rm. We conclude that malaria dynamics are indeed oscillatory when the methodology of explicitly incorporating the mosquito's demography, feeding and reproductive patterns is considered in modeling the mosquito population dynamics. A sensitivity analysis reveals important control parameters that can affect the magnitudes of Rm and R0, threshold quantities to be taken into consideration when designing control strategies. Both Rm and the intrinsic period of oscillation are shown to be highly sensitive to the mosquito's birth constant λm and the mosquito's feeding success probability pw. Control of λm can be achieved by spraying, eliminating breeding sites or moving them away from human habitats, while pw can be controlled via the use of mosquito repellant and insecticide-treated bed-nets. The disease threshold parameter R0 is shown to be highly sensitive to pw, and the intrinsic period of oscillation is also sensitive to the rate at which reproducing mosquitoes return to breeding sites. A global sensitivity and uncertainty analysis reveals that the ability of the mosquito to reproduce and uncertainties in the estimations of the rates at which exposed humans become infectious and infectious humans recover from malaria are critical in generating uncertainties in the disease classes.

Disruption of aminergic signalling reveals novel compounds with distinct inhibitory effects on mosquito reproduction, locomotor function and survival.

Insecticide resistance amongst disease vectors is a growing problem and novel compounds are needed. Biogenic amines are important for neurotransmission and we have recently shown a potential role for these in mosquito fertility. Here, we dissected the relative contribution of different aminergic signalling pathways to biological processes essential for vectorial capacity such as fertility, locomotion and survival by injecting agonists and antagonists and showed that octopaminergic/tyraminergic signalling is essential for oviposition and hatching rate. We show that egg melanisation is regulated by adrenergic signalling, whose disruption causes premature melanisation specifically through the action of tyramine. In addition to this, co-injection of tyramine with DOPA, the precursor of melanin, had a strong cumulative negative effect on mosquito locomotion and survival. Dopaminergic and serotonergic antagonists such as amitriptyline and citalopram recapitulate this effect. Together these results reveal potential new target sites for the development of future mosquito sterilants and insecticides.

AMPK and PTP‐1B stand at the gates of immunity, metabolism and egg formation in mosquitoes. (LB224)

Mosquitoes are major vectors of human diseases. This is mainly due to their requirement for a blood meal to lay eggs which allows the transmission of pathogens to the human being. Our aim is to identify proper molecular targets in order to block blood feeding and impair egg formation.In their early days of adult life mosquitoes feed on vegetal fluids and their midgut is continuously loaded with several bioactive compounds including polyphenols. We observed that polyphenol‐fed mosquitos dramatically increase their average lifespan (Control T 1/2 ± 15‐18 days, Polyphenol‐fed T1/2± 20‐32 days). Polyphenols also reduced the body triglyceride content from 20‐75 %. They trigger midgut autophagy which leads to a 40 % decrease on bacterial loads. Such mechanisms are mimicked by the activation of the mosquito AMPK and abolished by RNAi‐mediated silencing of this gene. Thus the presence of polyphenol on early mosquito meals optimizes mosquito metabolism and immunity and its chances of finding its first blood meal. After a blood meal digestion will take place in mosquito midgut. Blood‐derived nutrients allow yolk synthesis and egg formation. Blood feeding affects the expression of the mosquito PTP‐1B (AaPTP1B) in tissues critically involved in the synthesis and storage of vitellogenin (Vg), the major yolk protein. The protein tyrosine phosphatase inhibitor sodium orthovanadate or the PTP substrate DiFMUP decreased Vg synthesis and egg production. Also, silencing of AaPTP1B using by RNAi assays suppressed by 30 % egg production.In conclusion, the above results show that mosquito signaling pathways regulated by AMPK and PTP1B are proper targets to block blood feeding and egg formation. Future research strategies should be designed to generate novel cell signaling‐based insecticides that simultaneously decrease mosquito lifespan and reproduction.Grant Funding Source: Supported by CNPq, INCT‐EM, FAPERJ