L'évolution de la résistance aux insecticides chez les moustiques responsables de la transmission de maladies infectieuses compromet notre capacité à contrôler ces populations de vecteurs et pose de graves problèmes de santé publique. Mais les nombreuses modifications physiologiques associées au phénomène de résistance aux insecticides pourraient altérer l'épidémiologie de ces maladies de manière plus indirecte en modifiant la capacité vectorielle des moustiques. Afin d'étudier cette question nous avons mis en place un nouveau système expérimental composé du parasite aviaire Plasmodium relictum SGS1 et de son vecteur naturel le moustique Culex pipiens. Nous avons étudié l'effet de différents allèles de résistance aux insecticides (représentant deux mécanismes principaux i.e. la résistance métabolique ou la modification de la cible) sur une série de traits d'histoire de vie du parasite et du moustique. L'impact de ces différents allèles a été étudié d'une part, dans les conditions contrôlées de leur expression dans un même fond génétique (en utilisant plusieurs souches de moustiques isogéniques), et d'autre part, dans les conditions plus réalistes de leur expression dans un fond génétique hétérogène (utilisation de moustiques échantillonnés sur le terrain). Nous montrons que la résistance aux insecticides a des effets pleïotropes sur l'immunocompétence et les traits d'histoire de vie des moustiques. Son effet sur le développement de Plasmodium semble en revanche limité. Nous discutons d'une part, de la nécessité de poursuivre une approche multifactorielle (impliquant la physiologie, l'immunité et le comportement des moustiques) afin de mieux comprendre l'impact de la résistance aux insecticides sur la transmission de Plasmodium, et d'autre part des perspectives intéressantes qu'offrent ce nouveau système expérimental pour l'étude de l'écologie évolutive des maladies à vecteurs. / The evolution of insecticide resistance in mosquitoes threatens our ability to control many-vector-transmitted diseases, thereby raising serious public health issues. Insecticide resistance entails numerous physiological changes in mosquitoes. This thesis investigates whether these physiological changes alter the quality of mosquitoes as vectors of malaria. To address this issue, we developed a new experimental system consisting in the avian malaria parasite Plasmodium relictum SGS1 and its natural vector, the mosquito Culex pipiens. We investigated the impact of two insecticide resistance mechanisms (target site resistance and metabolic resistance) on several mosquito and parasite life history traits relevant for malaria transmission. The effect of different insecticide resistant genes was investigated using both isogenic laboratory mosquito strains (i.e. against a homogeneous genetic background) and sympatric field caught mosquitoes (i.e. under the more realistic, albeit noisier, conditions of a heterogeneous genetic background). We show that insecticide resistance has a pleiotropic effect on several mosquito traits (immunocompetence, longevity, fecundity), whereas it has only a limited effect on Plasmodium development. We discuss, on the one hand, the need to pursue such a multi-factorial approach (combining the mosquito physiology, immunity and behavior) to better understand the impact of insecticide resistance on malaria transmission and, on the other hand, the promising perspectives offered by this new experimental system for studying the evolutionary-ecology of infectious diseases.

This paper aims to analyze the correlation between the environmental, institutional and social with Infestation Index (II) at the neighborhoods Cidade Nova and Jabotiana in 2014. These districts were selected by virtue of respectively, presenting the lowest and highest II Aedes aegypti mosquito in Aracaju city. The research subjects were: six Municipal Health Department managers; Thirteen Fighting Endemic Diseases Agents (EDA) of Epidemiological Surveillance and two hundred residents of the neighborhoods studied. This is a descriptive study with qualitative and quantitative approach, conducted through the collection and analysis of data obtained by means of literature, documentary and field. In field research semi-structured interviews were conducted directed to managers of Epidemiological Surveillance and residents mentioned above. Interviews and workshops were also applied (Participatory Rapid Assessment - DRP) with the Agents. As from the systematization and analysis of the data were selected and measured environmental, institutional and social indicators that had relationship with the infestation of the Aedes aegypti. Thus, it was found that several factors can contribute to the discrepancy between the infestation levels in the analyzed districts, such as discontinuity and delay of actions to prevent and control the vector, lack of combat of EDA, precarious water supply system in some places, existence of areas with environmental vulnerability, with few green spaces and with the use and occupation of poorly planned ground. It was also observed that the institutional dimension showed that the worst rates in the studied neighborhoods and environmental dimension was the largest contributor to the sustainability of Jabotiana neighborhood. At the end of the research it was proposed management actions to confront the prevention difficulties and Aedes aegypti control, which may contribute to the municipal health management redirect their public policies, thus achieving more participatory and effective action. / Este trabalho objetiva analisar a correlação existente entre os indicadores ambientais, institucionais e sociais com o Índice de Infestação Predial (IIP) nos bairros Cidade Nova e Jabotiana, em 2014. Estes bairros foram selecionados em virtude de, respectivamente, apresentarem o maior e o menor IIP do mosquito Aedes aegypti no município de Aracaju. Os sujeitos desta pesquisa foram: seis gestores da Secretaria Municipal da Saúde; treze Agentes de Combate às Endemias (ACEs) da Vigilância Epidemiológica e duzentos moradores dos bairros estudados. Trata-se de um estudo descritivo com abordagem quali-quantitativa, realizado através da coleta e análise dos dados obtidos por meio de pesquisa bibliográfica, documental e de campo. Na pesquisa de campo foram realizadas entrevistas semiestruturadas direcionadas aos gestores da Vigilância Epidemiológica e aos moradores citados anteriormente. Foram também aplicadas entrevistas e oficinas (Diagnóstico Rápido Participativo - DRP) com os ACEs. A partir da sistematização e análise dos dados foram selecionados e mensurados indicadores ambientais, institucionais e sociais que possuíam relação com a infestação do mosquito Aedes aegypti. Assim, constatou-se que diversos fatores contribuem para a discrepância entre os índices de infestação nos bairros analisados, tais como descontinuidade e atraso das ações de prevenção e controle ao vetor, carência de Agentes de Combate às Endemias, precariedade no sistema de abastecimento hídrico em alguns locais, existência de áreas com vulnerabilidade socioambiental, com poucos espaços verdes e com um uso e ocupação do solo mal planejado. Observou-se ainda que, a dimensão institucional apresentou os piores índices para os bairros estudados e a dimensão ambiental foi a que mais contribuiu para a sustentabilidade do bairro Jabotiana. Ao final da pesquisa foram propostas ações de manejo para enfrentamento das dificuldades encontradas na prevenção e controle do Aedes aegypti, que poderão contribuir para que a gestão municipal da saúde redirecione suas políticas públicas, alcançando assim ações mais participativas e efetivas.

The Trypanosoma theileri group are blood parasites of ungulates. Confirmed vectors of these protists are horseflies or sheep keds for T. melophagium, but these trypanosomes were also detected in deer keds, mosquitoes and phlebotomus. We targeted on the investigation of possibility mosquitoes acting as vectors and additionally we targeted on the prevalence of T. theileri in mosquitoes at selected locations in the Czech Republic due to previous positive detection there. The measured prevalence has reached 6,4 % in mosquitoes, 4 % in deer keds and 16,7 % in sheep keds. The aim of experimental infections was to discover a potential of mosquitoes and phlebotomus acting as vectors of the T. theileri. Phlebotomus perniciosus and three species of mosquitoes Culex pipiens molestus, Culex pipiens quinquefasciatus and Aedes vexans were used for those experiments. Furthermore, isolates of T. theileri and T. melophagium from different sources were used. The divergence among these isolates and among used species of insect was found. The best results were achieved with Ae. aegypti and isolates from mosquitoes, where prevalence had reached 90,8 %. The infected mosquitoes were used for prediuresis research which had shown infection forms of the trypanosomes in the urine liquid. The phylogenetic analyse of T....

<p>  </p> <p>Vector-borne diseases (VBDs) represent a significant health burden worldwide, threatening approximately 80% of the global population. Insecticide-based vector control is the most effective method to manage many VBDs, but its efficacy has been declining due to high levels of resistance in vector populations to the main insecticide classes which operate via limited modes of action. Therefore, the discovery of new chemistries from non-conventional chemical classes and with novel modes of action is a priority for the control of vectors and VBDs. Natural products (NPs) are diverse in chemical structures and, potentially, modes of action. They have been used as insecticides for many decades and have inspired the development of multiple synthetic insecticides, suggesting the discovery of novel NPs could lead to the development of highly effective insecticides. </p> <p><br></p> <p>In this thesis, I report two studies with a main goal to identify novel mosquito-active insecticide leads that operate via modes of action distinct from existing insecticides. First, I tested the hypothesis that new mosquito-active insecticide leads with novel chemical structures, possibly operating via novel modes of action, can be identified by high-content larval phenotypic screening against a natural product collection and using novel phenotypic endpoints in addition to mortality endpoints. Here, I performed a high-content larval phenotypic screen using first instar (L1) larvae of <em>Aedes aegypti</em> (Linnaeus, 1762) against 3,680 compounds from the AnalytiCon MEGx Natural Product Libraries and a screening platform developed by Murgia et al., (2022). Compounds were screened in a 384-well plate format using the Perkin Elmer Opera Phenix and larvae were scored for lethal and novel phenotypic endpoints. Screening revealed five chemistries that caused larval mortality, including rotenone and a new NP chemistry, NP-4. The identification of rotenone confirmed the ability of the screen to detect mosquito-active NP chemistries. NP-4 caused high levels of larval mortality in the screen, and toxicity was confirmed in a subsequent concentration-response assay against third instar (L3) larvae of <em>Ae. aegypti</em>. 140 chemistries that caused atypical larval phenotypes, including cuticular pigmentation and morphometric changes relative to negative controls, were also identified by the screen. Some of these chemistries may operate by disruption of pathways regulating melanization, growth and development, and novel targets in the insect nervous systems, thus representing potential leads for further insecticide toxicity and mode of action studies. To facilitate quantitative analyses of atypical phenotypes, an attempt was made to assess the morphometrics of the thorax in larvae exposed to test chemistry, relative to control larvae. However, assessment was limited by the number of larvae images of suitable quality for measurements. </p> <p><br></p> <p>In the second study, I tested the hypothesis that metergoline (Murgia et al., 2022) and NP-4 (this study), two chemistries identified by the HTP phenotypic screen described in this project, operate via disruption of targets in the insect nervous systems that are distinct from the current insecticidal modes of products used in mosquito control programs. Specifically, I explored the hypothesis that metergoline operates via one or more insect orthologs of the mammalian G protein-coupled serotonin and dopamine receptors. An electrophysiology study was performed using the suction electrode technique and ganglia of the German cockroach, <em>Blattella germanica </em>(Linnaeus, 1767). To facilitate the investigation of metergoline agonism/antagonism and disruption of invertebrate GPCR signaling, 5-hydroxytryptamine (5-HT; serotonin) was included as a chemical probe. Electrophysiological recordings showed 5-HT (10µM and 1mM) and metergoline (10µM) caused no significant neurological activity at the tested concentrations in comparison to the saline control. However, a consistent neuro-inhibitory trend was observed, suggesting possible agonism of a 5-HT1-like receptor ortholog and antagonism of a putative 5-HT7-like receptor ortholog in the cockroach, respectively.  NP-4 caused significant neuro-inhibition at the tested concentration of 20µM, in comparison to the negative saline control. Given the demonstration of rapid contact toxicity to <em>Ae. aegypti</em> larvae and neurological inhibition in <em>B. germanica</em>, we propose NP-4 may act at one or more conserved targets in the insect nervous systems, which remain to be elucidated. </p> <p><br></p> <p>The significance of the present study is three-fold. First, this study reports the first high-content phenotypic screen of mosquito larvae against a NP collection and identification of 145 mosquito-active chemistries associated with lethal and phenotypic endpoints. These data confirm that the screening platform provided an innovative and effective system to rapidly identify mosquito-active small molecules with potential novel modes of action. Second, metergoline and NP-4 represent potential novel chemical leads for the development of new insecticides that can be incorporated into vector control programs targeting insecticide-resistant populations. Lastly, the study describes the first electrophysiology study of 5-HT, metergoline, and NP-4 via the suction electrode technique in an insect system and contributes new knowledge to the study of the insect serotonergic system, which represents an expanding area of vector biology research given its roles in feeding regulation.  </p> <p><br></p> <p>Future studies resulting from this thesis might include: (1) development of a set of morphometric criteria for quantitative analyses of atypical larval phenotypes, (2) incorporation of new phenotypic endpoints to expand the capacity of the screen to identify novel mode of action chemistries for insecticide discovery, and (3) identification of chemistry candidates suitable for further development from the 140 chemistries associated with atypical larval phenotypes in the primary screen using chemo-informatic and toxicological studies. In addition, studies using reverse transcription-polymerase chain reaction (RT-PCR), cell-based expression systems, mutant/insecticide resistant strains, and patch clamp electrophysiology could be pursued to further investigate the molecular mode of action of metergoline and NP-4, and potential for vector control.</p>

Malaria ist eine Krankheit, die durch den Protozoen Plasmodium verursacht und von Anopheles Moskitos durch infektiöse Stiche übertragen wird. Diese ̈Übertragung kann durch verschiedene Interventionsstrategien blockiert werden. Eine relativ neue Strategie, die bisher nur im Labor getestet wurde, ist der Einsatz genetisch veränderter Moskitos, die den Parasiten nicht auf einen neuen menschlichen Wirt übertragen können. Ein Ansatz ist die Entwicklung von Moskitos, die mit murinen Antikoepern ausgestattet sind, die gegen relevante Oberflächenproteine des Parasiten, dem Circumsporozoite Protein (CSP) gerichtet sind. Es ist jedoch nach wie vor unklar, welches Entwicklungsstadium angegriffen werden soll und welche Antikörper für diesen Ansatz effizient sind. Hier zeige ich, dass in Stechmücken, die mit einem humanen Anti-CSP-Antikörper ausgestattet sind, die Sporogonie der Oozysten in Abhängigkeit von der Parasiten-dichte blockiert wird und somit die Sporozoitenlast in den Mücken signifikant verringern. Insbesondere Antikörper, die sich an die ’Repeat Region’ des CSP binden, können die Sporozoitenlast in der Stechmücke verringern. Des Weitern, zeigen diese Stechmücken kaum Defekte in der Entwicklung und im Überleben. Diese Ergebnisse bestätigen die zuvor beschriebene Bedeutung von CSP während der Sporogonie und unterstreichen die Effizienz von humanen, ’Repeat Region’ bindenden Anti-CSP-Antikörpern bei der Beeinträchtigung der Parasitenentwicklung in dem Vektor. Darüber hinaus ist in Stechmücken, die mit humanen Anti-CSP Antikörpern ausgestattet sind, die Entwicklung von Sporozoiten teils limitiert und teils komplett verhindert. Dies macht sie zu einem vielversprechenden Instrument für Maßnahmen zum Malaria Kontrolle. In dieser Arbeit habe ich weitere Einblicke in den Mechanismus , durch den Anti-CSP-Antikörper die Parasitenentwicklung in der Mücke stören, und gezeigt, dass Oozysten ein effizientes Ziel für diesen Ansatz sind. / Malaria is a disease caused by the protozoan parasite Plasmodium and transmitted by Anopheles mosquitoes trough infectious bites, these transmission events can potentially be blocked by different intervention strategies. A relatively new strategy which has been only tested in the laboratory is the use of genetically modified mosquitoes unable to transmit the parasite to a new human host. In the past, murine derived antibodies directed against relevant parasite surface proteins were used with limited success. It remains unclear which developmental stage is targeted, and which antibodies are useful. Here, I show that in mosquitoes equipped with a human derived anti-CSP repeat binding antibody, oocyst sporogony is blocked in a parasite density dependent manner. Only repeat binding antibodies could decrease sporozoite loads in the mosquito. These results confirm the previously described importance of CSP during sporogony and highlight the efficiency of human derived repeat binding anti-CSP antibodies in interfering with parasite development even in a different host. Additionally, mosquitoes equipped with human derived anti-CSP antibodies show little (in high density infections) to no (in low density infections) sporogonic development, making them a promising tool for malaria transmission blocking interventions. I provided additional insights into the mechanism by which anti-CSP antibodies interfere with parasite development in the mosquito showing that oocysts are an efficient target for this approach. Therefore, the mosquitoes used here are potentially resistant in a more natural setting. Additionally, I provided a new tool allowing a faster screening of antibodies in a mosquito context by injection of single chain Fabs into the mosquito hemolymph. Taken together, this approach could one day give rise to alternative strategies in tackling malaria transmissions.

A population survey was conducted on the mosquito species recorded in Denton, Texas for the years of 2005 to 2015. Data used in this project were obtained from an ongoing, long-term surveillance program led by the City of Denton and conducted through the University of North Texas. Research focused on the population dynamics and community structure of mosquitoes collected within urban areas of Denton, Texas in relation to certain environmental variables. A total of 80,837 female mosquitoes were captured and represented 38 species found under the following genera: Aedes, Anopheles, Coquillettidia, Culex, Culiseta, Mansonia, Orthopodomyia, Psorophora, Toxorhynchites, and Uranotaenia. Culex quinquefasciatus was the most abundant species followed by Aedes vexans. Seasonal patterns of the most abundant species revealed high variability throughout the study. Container breeders were most abundant in August and those that breed in floodwaters were most abundant in the months of May and September. Samples were tested for arbovirus presence through the Texas Department of State Health Services in Austin, Texas and multiple pools tested positive for West Nile virus throughout the study. Stepwise multiple regression and Spearman's rank correlation analyses were performed to examine the relationship between the mosquito community and environmental variables. Data revealed that temperature, precipitation, and dew point were the most important variables influencing the mosquito population in the City of Denton.

Insects maintain blood (haemolymph) Ca2+ concentrations within a narrow range in order to support the health of internal tissues and organs. The Malpighian (renal) tubules play a primary role in haemolymph Ca2+ homeostasis by sequestering excess Ca2+ within calcified biomineral deposits (Ca-rich granules) often located within type I (principal) tubule cells. Using the classic Ramsay assay, the scanning ion-selective microelectrode technique (SIET), and modifications of these two electrophysiological techniques, this thesis begins to unravel the sites and mechanisms of Ca2+ transport by the Malpighian tubules isolated from eight insects, representing seven orders. A segment-specific pattern of Ca2+ flux was observed along the length of the Malpighian tubules isolated from D. melanogaster, A. aegypti and A. domesticus and was uniform along the length in the remaining species. The majority (≥ 90%) of Ca2+ entering the tubule cells is sequestered within intracellular calcium stores in Ca2+-transporting segments of D. melanogaster and A. domesticus tubules, consistent with the presence of Ca-rich storage granules in these tubule segments. In addition, this thesis provides the first measurements of basolateral Ca2+ flux across single principal and secondary tubule cells of T. ni, where Ca2+ uptake occurs only across principal cells. Perhaps the most important finding of this thesis is that increasing fluid secretion through manipulation of intracellular levels of cAMP or Ca2+ in isolated tubules of A. domesticus had opposite effects on tubule Ca2+ transport. The adenylyl cyclase-cAMP-PKA pathway promotes Ca2+ sequestration whereas both 5-hydroxytryptamine and thapsigargin inhibited sequestration. In contrast, tubules of the remaining species were generally insensitive to cAMP or thapsigargin and v rates of tubule Ca2+ transport were often very low. The presence of Ca-rich granules in the cells of the midgut in several of the species with low rates of tubule Ca2+ transport provide evidence for a putative role of the midgut in haemolymph Ca2+ homeostasis. Taken together, these results suggest that the principal cells of the Malpighian tubules contribute to haemolymph calcium homeostasis through neuroendocrine regulated sequestration of excess Ca2+ during periods of high dietary calcium intake. Sequestration of dietary Ca2+ by the midgut may reduce Ca2+ entry into the haemolymph and therefore Ca2+ sequestration by the Malpighian tubules need not be so rapid. Finally, reversible tubule Ca2+ transport may allow internal reserves of Ca2+ (Ca-rich granules) to be returned to the haemolymph allowing insects to survive prolong periods of Ca2+ deficiency (i.e. overwintering). / Thesis / Doctor of Philosophy (PhD) / This thesis contributes to our understanding of how insects regulate the calcium content of their blood (haemolymph). Using electrophysiological techniques with improved spatial resolution (from millimeters to micrometers) this thesis sought to determine the sites, mechanisms and regulation of Ca2+ transport by insect Malpighian (renal) tubules in order to gain insights into the role of Ca-rich granules (similar to those identified in early stages of human kidney stone formation i.e. nephrolithiasis) within these tissues. Using eight insect species this thesis demonstrates that the Malpighian tubules act as dynamic Ca2+ stores that appear to be under neuroendocrine control: actively taking up Ca2+ through calcium entry channels, where the majority (≥ 90%) of excess haemolymph Ca2+ is sequestered within intracellular stores (Ca-rich granules) during period of excess dietary calcium and passively releasing Ca2+ back to the haemolymph during periods of metamorphosis or calcium deficiency (i.e. overwintering).

Le paludisme, qui est la maladie à transmission vectorielle la plus répandue, provoque depuis quelques années de plus en plus d’épidémies liées à des anomalies climatiques. Dans de nombreux pays endémiques comme l’Ouganda, les changements climatiques représentent une préoccupation importante pour la santé publique. Des débats existent toutefois quant à l’évolution future du paludisme car la majorité des études de prédictions ne considèrent pas les effets de certains facteurs anthropiques qui influencent la transmission (ex. les interventions de contrôle antivectorielles). Ainsi, les objectifs de cette thèse étaient donc 1) d’estimer les associations entre le risque du paludisme, des variables de l’environnement (comme les précipitations, la température, l’humidité et la végétation) et les interventions antivectorielles (moustiquaires imprégnées d’insecticide longue durée, MILD, et pulvérisation intra-domiciliaire, PID) pour 2) prédire la distribution du paludisme selon des scénarios de climat futur. À cette fin, les associations ont été étudiées à partir (i) des données d’une cohorte d’enfants de trois sous régions d’Ouganda à partir de modèles mixtes linéaires généralisés basés sur une distribution log-binomiale; (ii) des données de surveillance passive du paludisme dans la population générale de six sous régions, à partir de modèles mixtes linéaires généralisés basés sur une distribution binomiale négative. Les associations étudiées au sein de la population générale ont ensuite été utilisées pour projeter le risque futur selon 14 simulations climatiques et deux scénarios d’émission de gaz à effet de serre (RCP4.5 et RCP8.5). Pour le premier objectif, les résultats de l’analyse des données de la cohorte infantile ont mis en évidence une variabilité sous régionale dans la forme (linéaire et non linéaire), la direction et l'ampleur des associations entre les variables de l’environnement et le risque de paludisme. L'ajustement du modèle de régression pour la PID a modifié l'ampleur et/ou la direction des associations environnement-paludisme, suggérant un effet d'interaction. À partir des données de la population générale, l’analyse groupée des six sous régions a montré que les interventions réduisaient le risque de paludisme d'environ 35 % avec les MILD et de 63 % avec la PID; des interactions significatives ont été observées entre certaines variables environnementales et les interventions de lutte antivectorielle. À l’échelle sous régionale, une variabilité de la forme des relations environnement-paludisme (linéarité, non linéarité, direction) et de l'influence des interventions a aussi été observée. Les prédictions du risque de paludisme avec les changements climatiques suggèrent des tendances à la hausse des cas de paludisme en absence d’interventions à l’horizon 2050, bien qu’une grande variabilité dans les prédictions existent selon le modèle de climat considéré (médianes et min-max de la période historique vs RCP 4.5 : 16 785, 9 902 - 74 382 vs 21 289, 11 796 - 70 606). En considérant l’effet des interventions, une réduction du nombre de cas annuels médian de 35%, 63% et à 76% est prédite avec les MILD seules, la PID seule et la combinaison de MILD et PID, respectivement. Cette thèse a donc permis de clarifier l’influence des MILD et de la PID sur les relations entre variables de l’environnement et le paludisme et de démontrer l’importance de considérer les mesures de contrôle antivectorielle dans les analyses du risque épidémiologique de paludisme et dans les prédictions du risque avec les changements climatiques. / Malaria, which is the most widespread vector-borne disease, has in recent years caused more and more epidemics linked to climatic anomalies. In several malaria endemic countries such as Uganda, climate change is a major public health concern. Debates exist, however, about the future evolution of malaria in relation to climate as the majority of prediction studies do not consider the effects of certain anthropogenic factors that influence transmission (e.g. vector control interventions). Therefore, the objectives of this thesis were 1) to estimate the associations between malaria risk, environmental variables (such as precipitation, temperature, humidity which are related to climate and vegetation) and vector interventions (long-lasting insecticide-treated bed nets - LLINs, and indoor residential spraying - IRS), and to 2) predict malaria distribution under future climate scenarios. The associations were studied with (i) data from a cohort of children from three sub-regions of Uganda using generalized linear mixed models based on a log-binomial distribution; (ii) data from passive surveillance of malaria in the general population of six sub-regions, using generalized linear mixed models based on a negative binomial distribution. The associations studied in the general population were then used to predict future risk under 14 climate simulations and two greenhouse gas emission scenarios (RCP4.5 and RCP8.5). For the first objective, the results of the analysis of the infant cohort data highlighted a sub-regional variability in the form (linear and nonlinear), the direction and the magnitude of the associations between the environmental variables. and the risk of malaria. Adjusting the regression model for IRS changed the magnitude and/or direction of environment-malaria associations, suggesting an interaction effect. Using data from the general population, the pooled analysis of the six sub-regions showed that the interventions reduced malaria risk by approximately 35% with LLINs and by 63% with IRS; significant interactions were observed between some environmental variables and vector control interventions. At the sub-regional scale, variability in the form of environment-malaria relationships (linearity, non-linearity, direction) and in the influence of interventions was also observed. Predictions of malaria risk with climate change suggest upward trends in malaria cases in the absence of interventions by 2050, although great variability in the predictions exists depending on the climate model considered (medians and min-max of the historical period vs RCP 4.5: 16,785, 9,902 - 74,382 vs 21,289, 11,796 - 70,606). Considering the effect of interventions, a reduction in the median number of annual cases of 35%, 63% and 76% is predicted for LLINs alone, IRS alone and the combination of LLINs and IRS, respectively. This thesis examined the influence of LLINs and IRS on the relationships between environmental variables and malaria and demonstrates the importance of considering vector control measures in analyzes of the epidemiological risk of malaria and in its prediction with climate change.

Einer der ausschlaggebenden Faktoren für das Übertragungspotenzial von Malaria ist die Häufigkeit ihres Überträgers, der Anopheles-Mücke. Da diese Insekten besonders empfindlich auf ihre Umgebung reagieren, wurde in vielen Laborexperimenten das Verhältnis zwischen Umweltfaktoren und Malaria untersucht. In natürlichen Umgebungen ist über diesen Zusammenhang wenig bekannt. Untersuchungsgegenstand dieser Arbeit mit multidisziplinärem Ansatz ist die Beziehung zwischen verschiedenen Umwelteinflüssen und der Häufigkeit von Anopheles-Mücken in natürlicher Umgebung sowie die möglichen diesem Prozess zugrundeliegenden Mechanismen. Zunächst lässt sich anhand von Feldstudien und empirischen dynamischen Modellen eine verzögerte Auswirkung der Umwelt auf die Mückenhäufigkeit feststellen, was wiederum einen Einfluss auf die Entwicklung der Larvenstadien bedeuten könnte. Im Anschluss werden mögliche Hintergründe untersucht mit Hilfe eines kontrollierten Versuchsaufbaus, in dem Anopheles-coluzzii-Larven und Erwachsenentiere auf Umweltbedingungen reagieren. Dabei lässt sich feststellen, dass Temperatur und Besatzdichte vielschichtige Auswirkungen auf Larvenentwicklung und -sterblichkeit haben. Darüber hinaus gibt es Übertragungseffekte auf die Alterung und Fruchtbarkeit der Erwachsenentiere. Abschließend zeigt sich durch die Entwicklung eines vollständigen Modells für den Lebenszyklus von Anopheles-Mücken, dass die ermittelten Umwelteinflüsse auf Larven und erwachsene Mücken einen komplexen Einfluss auf die Größe der Mückenpopulation haben. Anhand mehrere mathematischer Methoden werden in dieser Arbeit Informationen aus Feldsammlungen und Versuchsdaten erfolgreich kombiniert. Dies unterstreicht die Bedeutung multidisziplinärer Ansätze für das Verständnis der komplexen Dynamik zwischen Mücken und der Umwelt. So entsteht ein detailliertes Bild der Lebensmerkmale der Mücken, die von der Umwelt beeinflusst werden, und deren Auswirkungen auf die Populationsdynamik. / One of the main determinants of malaria transmission is the abundance of its vector, the Anopheles mosquito. As these insects are susceptible to the environment in which they live, many laboratory experiments have explored the connection between environmental factors and the abundance of mosquitoes. However, little is known about this relationship in natural settings. In this work, I used a multidisciplinary approach to determine the relationship between environmental drivers and Anopheles abundance and the possible mechanisms that characterise these relationships. First, using field study data and an empirical dynamic modelling approach, I identified a delayed effect of the environment on mosquito abundance, suggesting an impact on the development of larval stages. Second, I explored the possible mechanisms behind this process using a controlled experimental setup in which Anopheles coluzzii larvae and adults responded to the environmental conditions. I identified a complex response to temperature and rearing density of larval development and mortality, as well as carry-over effects on adult ageing and fecundity. Finally, by developing a full model for the life cycle of Anopheles mosquitoes, I determined that the identified environmental impacts on larvae and adult mosquitoes have a complex impact on the mosquito population size. Taken together, this work combined the information from field collections and experimental data successfully using multiple mathematical methods, highlighting the importance of multidisciplinary approaches in understanding the complex dynamics between mosquitoes and the environment. With this approach, we obtained a detailed picture of the mosquito life traits that are impacted by the environment and its effects on mosquito population dynamics.

Avian malaria (Plasmodium spp.) is a popular model system to study the ecology and evolution of parasite-host-vector interactions in the wild. These studies have historically focused mostly on the avian hosts and the malaria parasites. Knowledge regarding the role of vectors is essential to our understanding of these wild systems, but has only very recently started to accumulate. This thesis aimed to contribute to this field by assessing mosquito-malaria-host associations for British mosquitoes and the role of mosquito ecology in shaping these parasite systems in a British woodland study site, using molecular, field ecology and statistical modelling methodologies. From the 12 mosquito species or species groups found, I showed that the Cx.pipiens/torrentium mosquito group is likely to have a major role in avian malaria transmission in Great Britain, while Cs. annulata may be transmitting P. circumflexum. I also demonstrated a positive spatial association between mosquito density per host and avian malaria prevalence, in accordance with theoretical expectations for malaria transmission. Findings here provide evidence that avian malaria transmission in British woodlands is limited mainly to June-August, being preceded by relapse of previous infections or, alternatively, by maintenance of chronic blood parasitaemia through the colder months; this agrees with theoretical expectations and findings elsewhere for temperate climates. This thesis also described local-scale spatial heterogeneity and seasonal variation in adult mosquito abundance within a British woodland where avian malaria is endemic, with differing patterns found between species or species groups. Spatially, variation in adult mosquito abundance was associated with microclimatic and landscape variables such as distances to mosquito breeding sites, microclimate and canopy height; seasonally, variation in mosquito abundance was associated with temperature and rainfall, alongside calendar date. The heterogeneity in mosquito parameters and associations with environmental variables found at a site where avian malaria is endemic highlights the need to anticipate such complexity when trying to understand Plasmodium transmission. By doing so, we further extend the potential of these parasite systems to improve our knowledge regarding the ecology and evolution of parasite-host-vector associations.