Few studies regarding treatment seeking behaviour related to Malaria have been conducted in Uganda, and most of the studies regarding use of mosquito nets that have been conducted are quantitative. There is thus a need for qualitative studies for a deeper understanding of the complex issue of coping with malaria in a resource limited setting. The aim of this study is to understand sociocultural and structural factors influencing the coping with malaria in rural Uganda. Focus is on the local experiences, perceptions and hardships regarding seeking treatment for malaria and preventative measures like the use of mosquito nets. A theoretical framework based on Political ecology of health, with a focus on human agency, is used to analyse these issues. The village Nyakasojo in Kasese district was selected for a field study. The main source of information was open-ended interviews conducted in March 2020. The study showed that the main reason for not using mosquito nets was lack of access to nets and easily torn nets, rather than unwillingness to use mosquito nets. Further, the study showed that the unequal access to healthcare in combination with livelihood vulnerability forces people to wait before seeking treatment and/or taking half doses of antimalarials, probably leading to unnecessary malaria transmission. A more holistic approach to malaria control would be beneficial, including the aspect of treatment in addition to the present narrower focus on prevention.

Filarial parasites are a type of nematode that requires arthropod vectors for transmission between hosts. Filarial parasites vary among species of vertebrate hosts and can cause varying symptoms in hosts, including death. The presence of filarial parasites can influence host populations and can be costly to infected areas. To evaluate the prevalence of filarial parasites in Lake County, CA, mosquitoes were collected in 2014 and analyzed for infection using polymerase chain reaction (PCR). Of 1,008 mosquito pools, six filarial parasite species were detected in 23 pools representing six mosquito species. The DNA of Dirofilaria immitis (n=6, MIR=0.26); Setaria yehi (n=9, MIR=1.44); Splendidofilaria sp. (n=4, MIR=0.20); unknown filarial parasites A (n=2, MIR=0.09), B (n=1, MIR=0.41), and C (n=1, MIR=0.05) were detected in Aedes increpitus, Aedes sierrensis, Anopheles franciscanus, Anopheles freeborni, Culex stigmatosoma, and Culex tarsalis mosquito pools. Due to the presence of D. immitis, which can lead to chronic illness and death in domestic dogs, in Lake County it is important to evaluate vector competency of D. immitis in mosquito species. Culex tarsalis was chosen due to the high abundance found in Lake County in 2014 (n=36,587). To evaluate vector competency of Cx. tarsalis in transmission of D. immitis, colony and field-caught Lake County (n=102, n=54 respectively) mosquitoes were analyzed for infectivity using decapitation. Fourteen days post feeding on infected blood, mosquitoes were decapitated to evaluate the presence of L 3 larvae; but no L 3 larvae were detected. The presence of D. immitis DNA was detected in eight colony (IR=7.8%) and fifteen field-caught (IR=23.1%) thoraces using PCR. Though no L 3 larvae were observed in decapitated mosquitoes, presence of D. immitis DNA in the thoraces of mosquitoes using PCR has previously been used as an indicator for vector competency. Thus it is probable that Cx. tarsalis is a competent vector for D. immitis.

No description available.

Alphaviruses are a group of medically relevant arthropod-borne viruses (arboviruses) belonging to the Togaviridae family that are maintained by mosquito vectors. These zoonotic viruses are clustered into two groups: New World and Old World, depending on their geographical origin/distribution and clinical manifestations. Both of these groups cause disease symptoms of an acute febrile illness; however, each group has a distinct, hallmark disease symptom; New World alphaviruses, such as Eastern, Western, and Venezuelan equine encephalitis viruses (EEEV, WEEV, and VEEV, respectively), present with severe encephalitis while Old World alphaviruses, such as Sindbis, chikungunya, and Mayaro viruses (SINV, CHIKV, and MAYV, respectively) present with an incapacitating polyarthralgia that can persist for years following initial infection. To date, the most effective means of controlling these arboviral infections is through mosquito control programs. However, these programs have crucial limitations in their effectiveness; therefore, novel approaches are necessary to control the spread of these crippling pathogens and lessen their disease burden. Given the close phylogenetic and antigenic relationship between MAYV and CHIKV, we hypothesized that prior CHIKV immunity may affect the outcome of MAYV disease and/or limit its emergence in humans. Our work has shown that anti-CHIKV neutralizing antibodies can provide cross-protective immunity against MAYV disease. Alongside these studies, we have characterized the potency of a camelid-derived single-domain antibody (sdAb) that neutralizes a breadth of alphaviruses, including CHIKV and MAYV. With these data, we have designed and generated transgenic Aedes aegypti mosquitoes that express two anti-CHIKV sdAbs to target infection, dissemination, and transmission of MAYV and CHIKV within this deadly vector. These findings are particularly significant because they highlight the ability to co-target two emerging alphaviruses that are crippling public health and obliterating quality of life around the globe within a single defense strategy. / Doctor of Philosophy / Alphaviruses are arthropod-borne viruses (arboviruses) belonging to the Togaviridae family that infect millions of people annually via the bite of female mosquitoes. These viruses are major public health threats due to their ability to infect humans and animals and infections resulting in a range of debilitating diseases. Viruses within this genus are clustered into two groups: Old World and New World, based on geographical origin and distribution. While New World alphaviruses are known for inducing severe encephalitis (i.e., swelling in the brain), a hallmark symptom of the Old World alphaviruses is the development of incapacitating polyarthralgia (i.e., widespread joint pain) that can persist for years following initial infection. To date, the most effective means of combatting these viruses is through mosquito control programs. However, these programs have crucial limitations in their effectiveness; therefore, novel approaches are necessary to control the spread of these crippling pathogens. Given the close genetic relationship between chikungunya virus (CHIKV) and Mayaro virus (MAYV), our research has focused on harnessing cross-reactive immunity between these emerging alphaviruses. We discovered this cross-reactivity provides protective immunity to both viruses (i.e., CHIKV and MAYV) after exposure to only one (i.e., CHIKV) of the viruses. Next, we characterized the potency of a small, single-domain antibody (sdAb) to neutralize a breadth of alphaviruses, including CHIKV and MAYV. With these data, we have designed and generated transgenic Aedes aegypti mosquitoes that express this sdAb to target both CHIKV and MAYV within this deadly mosquito vector. These findings are particularly significant because they provide the foundation for a novel approach to controlling and preventing outbreaks of these emerging alphavirus pathogens that obliterate quality of life in public health settings around the globe.

The influence of forest canopy disturbance (FCD) on La Crosse virus (LACV), leading cause of US pediatric arboviral encephalitis, is critical to understand in landscapes where forests are periodically harvested. Southwestern Virginia is part of an emerging focus of this interior forest bunyavirus. I investigated how the temperate forest mosquito community, LACV vectors, and the LACV amplifying vertebrate host (chipmunks) were impacted by logging. This research was conducted across an experimental FCD gradient (from least to most disturbed: contiguous control, fragmented control, clearcut, and high-leave shelterwood (SW)). Using gravid traps, I found that the mosquito community was resilient to logging with no significant difference in diversity or community composition across treatments. Mean number of female mosquitoes caught per trap-night declined with disturbance. FCD significantly affected the abundance of vector species in different ways. The primary LACV vector, Aedes triseriatus, and the recent invasive Ae. japonicus declined with logging. Other vectors (Ae. albopictus, Ae. canadensis, and Ae. vexans) thrived with logging. Culex pipiens/restuans was affected by disturbance but had no treatment preference. A mark-recapture study revealed that chipmunk abundance and LACV seroprevalence were greatest on the SW. In sync with Ae. triseriatus abundance but in contrast to the chipmunk results, mosquito LACV detection was significantly greater on unlogged sites. Surprisingly, LACV was detected in Ae. japonicus and Cx. pipiens/restuans. In a follow-up study, I isolated LACV from field-collected Ae. japonicus. Although LACV was previously isolated from Cx. pipiens, the vector competence was unknown. Therefore, I examined the vector competence of Cx. pipiens and Cx. restuans. Although poor vectors, I did detect LACV in the saliva of both species. An additional experiment found that nutritionally-stressed Cx. restuans were better vectors than those in the control group, indicating that environmental stressors (e.g., FCD) may alter the ability of accessory vectors to spread LACV. The influence of FCD on LACV is complex. Because logging decreases Ae. triseriatus abundance, human LACV risk is likely lowered by decreased transovarial vertical transmission. However, high chipmunk seroprevalence on disturbed sites suggest horizontal transmission with accessory vectors plays a larger role in LACV risk on recently logged sites. / Ph. D.

The transcription factor Krüppel homolog 1 (Kr-h1) is crucial in multiple reproductive processes of Aedes aegypti mosquitoes, including previtellogenesis, vitellogenesis, and oogenesis. This study explores the interaction between Kr-h1 and its potential associated proteins (KAPs), with a specific focus on the dimerization partner (DP-1), and how this interaction regulates gene expression pathways critical for mosquito reproduction. Utilizing RNA interference (RNAi), the research identifies DP-1 as a significant regulator of follicle growth post-eclosion (PE), highlighting its vital role in the mosquito reproductive regulatory pathway. The experimental approach included RNAi-mediated knockdown of DP-1, accompanied by evaluations using quantitative PCR (qPCR), Western blotting (WB), co-immunoprecipitation (Co-IP), follicle length measurement, and egg counting to assess the role of DP-1 in reproductive functions. For the first time, the inhibition of DP-1 expression was found to significantly impede A. aegypti follicular development. The elucidation of the mechanistic roles of Kr-h1 and DP-1 provides valuable insights that could lead to innovative strategies for mosquito population control and effective disease vector management. / Master of Science in Life Sciences / Mosquitoes can spread serious insect-borne diseases such as dengue, Zika, and malaria, etc. These diseases can infect hundreds of millions of individual and cause around a million of death annually. This study focuses on a specific mosquito species, Aedes aegypti, which is a major carrier of these diseases. To manage their populations and reduce the spread of these diseases, scientists are constantly seeking new methods to control their reproduction. Chemical insecticides are one of the most efficient and widely used strategies. However, these insecticides face significant challenges, including the development of resistance in mosquito populations and the potential damage to non-target species to affect the ecosystem. To address this issue, the development of new insecticides is crucial. We can identify new targets to pave the way to research novel effective insecticides. Inside mosquitoes, there are various proteins that help control their ability to reproduce. One of these proteins is called Krüppel homolog 1 (Kr-h1). Kr-h1 plays a crucial role in the development and reproductive processes of mosquitoes. Our research looked at how Kr-h1 interacts with other types of protein to control mosquito reproduction. Through various experiments, including gene expression analysis and protein studies, we found that DP-1 is essential for the proper development of mosquito eggs. This insight helps us understand more about the biological processes and hormonal pathways during mosquito reproduction, therefore provide greater opportunity to develop insecticides to reduce their populations and the spread of the diseases they carry.

Diseases such as malaria and dengue impose great economic burdens and are a serious threat to public health, with young children being among the worst affected. These diseases are transmitted by mosquitoes, also called disease vectors, which are able to transmit both parasitic and viral infections. One of the most important strategies in the battle against mosquito-borne diseases is vector control by insecticides and the goal is to prevent people from being bitten by mosquitoes. Today’s vector control methods are seriously threatened by the development and spread of insecticide-resistant mosquitos warranting the search for new insecticides. This thesis has investigated the possibilities of vector control using non-covalent inhibitors targeting acetylcholinesterase (AChE); an essential enzyme present in mosquitoes as well as in humans and other mammals. A key requirement for such compounds to be considered safe and suitable for development into new public health insecticides is selectivity towards the mosquito enzyme AChE1. The work presented here is focused on AChE1 from the disease transmitting mosquitoes Anopheles gambiae (AgAChE1) and Aedes aegypti (AaAChE1), and their human (hAChE) and mouse (mAChE) counterparts. By taking a medicinal chemistry approach and utilizing high throughput screening (HTS), new chemical starting points have been identified. Analysis of the combined results of three different HTS campaigns targeting AgAChE1, AaAChE1, and hAChE allowed the identification of several mosquito-selective inhibitors and a number of compound classes were selected for further development. These compounds are non-covalent inhibitors of AChE1 and thereby work via a different mechanism compared to current anti-cholinergic insecticides, whose activity is the result of a covalent modification of the enzyme. The potency and selectivity of two compound classes have been explored in depth using a combination of different tools including design, organic synthesis, biochemical assays, protein X-ray crystallography and homology modeling. Several potent inhibitors with promising selectivity for the mosquito enzymes have been identified and the insecticidal activity of one new compound has been confirmed by in vivo experiments on mosquitoes. The results presented here contribute to the field of public health insecticide discovery by demonstrating the potential of selectively targeting mosquito AChE1 using non-covalent inhibitors. Further, the presented compounds can be used as tools to study mechanisms important in insecticide development, such as exoskeleton penetration and other ADME processes in mosquitoes.

A population survey was conducted from April through September 2002 on mosquito species occurring on the Ray Roberts Greenbelt, a riparian corridor used for public recreation on the Elm Fork of the Trinity River, in Denton County, Texas. ArcGIS software was used to set up a stratified random sampling design based on habitat parameters. Multivariate analyses of sampling data and climatic variables were used to describe spatial and temporal patterns of mosquito species. A total of 33 species were collected during this study belonging to the following genera: Aedes, Anopheles, Coquillettidia, Culex, Mansonia, Ochlerotatus, Orthopodomyia, Psorophora, Toxorhynchites, and Uranotaenia. Seasonal distributions of the dominant species revealed population fluctuations. Aedes vexans was the primary species collected in April and May, occurring in low numbers throughout the rest of the sampling period. Psorophora columbiae reached its highest population density in June, with a smaller peak occurring in late July. Present from May through the end of September, Culex erraticus was the most abundant species collected with major peaks in mid-June and the end of July. Abundance of Culex salinarius followed the same general trend as that for Cx. erraticus, but with smaller numbers. The specimens were tested for a variety of arboviruses by the Texas Department of Health. One pool of Cx. erraticus and Cx. salinarius, collected in August 2002, tested positive for West Nile virus. Variables that were important factors for determining dominant species abundance were temperature, wind speed, rain accumulation occurring one-week and two-weeks prior to sampling, number of day since last rain event, dew point, and average canopy coverage.

A number of emerging vector-borne diseases have the potential to be transmitted in the UK by native mosquitoes. Human infection by some of these diseases requires the presence of communities of multiple vector mosquito species. Mitigating the risk posed by these diseases requires an understanding of the spatial distributions of the UK mosquito fauna. Little empirical data is available from which to determine the distributions of mosquito species in the UK. Identifying areas at risk from mosquito-borne disease therefore requires statistical modelling to investigate and predict mosquito distributions. This thesis investigates the distributions of potential vector mosquitoes in the UK at landscape to national scales. A number of new methodological approaches for species distri- bution modelling are developed. These methods are then used to map and understand the distributions of mosquito communities with the potential to transmit diseases to humans. Chapter 2 reports the establishment of substantial populations of the West Nile virus (WNV) vector mosquito Culex modestus in wetlands in southern England. This represents a drastic shift in the species’ known range and an increase in the risk of WNV transmission where Cx. modestus is present. Chapter 3 develops and applies a new species interaction distribution model which identifies fish and ditch shrimp of the genus Palaemonetes as predators which may restrict the distribution of the potential WNV vector community in these wetlands. Chapter 4 develops a number of methods to make robust predictions of the probability of presence of a species from presence-only data, by eliciting and applying estimates of the species’ prevalence. Chapter 5 introduces a new Bayesian species distribution modelling approach which outperforms existing methods and has number of useful features for dealing with poor- quality data. Chapter 6 applies methods developed in the previous two chapters to produce the first high-resolution distribution maps of potential vector mosquitoes in the UK. These maps identify several wetland areas where vector communities exist which could maintain WNV transmission in birds and transmit it to humans. This thesis makes significant contributions to our understanding of the distributions of UK mosquito species. It also provides methods for species distribution modelling which could be widely applied in ecology and epidemiology.

Doctor of Philosophy / Department of Chemistry / Duy H. Hua / Our research work is divided into three chapters. In the first chapter, synthesis of substituted phenolic compounds including halogenated di- and trihydroxybenzenes, aminophenols, and substituted di-tert-butylphenols, their redox potential, laccase oxidation, and mosquito anti-larval activities are discussed. The synthesized substituted phenols were found to be the substrates but not the inhibitors of laccase. An inverse correlation between the oxidation potential and the laccase oxidation efficiency of halogenated hydroxybenzenes and aminophenols was established. However, substituted di-tert-butylphenols were found to have anti-larval activities in mosquitoes resulting in the death of the larvae just before reaching pupation. Among the di-tert-butyl phenols studied, water insoluble, 2,4-di-tert-butyl-6-(3-methyl-2-butenyl)phenol (16), 2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-methylpropanal oxime (14), and 6,8-di-tert-butyl-2,2-dimethyl-3,4-dihydro-2H-chromene (17) caused the mortility of 98%, 93%, and 92% of Anopheles gambiae larvae in the concentration of 182 nM, 3.4 µM, and 3.7 µM, respectively. In particular, compound 16 had similar anti-larval activities as compared to MON-0585, an anti-larval agent reported by Monsanto in the 70’s. In the second chapter, inhibition of protein kinase C (PKC) phosphorylation by substituted quinolines (PQs) is inverstigated. PQ compounds such as N-(3-aminopropyl)-6-methoxy-4-methyl-5-(3-(trifluormethyl)phenoxy)quinolin-8-amine (PQ1), N-(furan-2-ylmethyl)-6-methoxy-4-methyl)-5-(3-(trifluoromethyl)phenoxy)quinolin-8-amine (PQ11), and 6-methoxy-4-methyl-N-(quinolin-4-ylmethyl)-5-(3-(trifluoromethyl)phenoxy)quinolin-8-amine (PQ15) were found to inhibit PKC phosphorylation with IC50 values of 35 nM, 42.3 nM, and 216.3 nM respectively, among which PQ1 and PQ11 were found to be potent PKC inhibitors as comparable to that of staurosporine (IC50 = 33 nM). In chapter three, the tissue distribution of PQ1 and PQ11 in normal C57BL/6J mice and the effect of PQ1 on the normal tissues of mice were investigated. Substituted quinolines, PQ1 and PQ11 were distributed in the tissues in concentrations that were more than 40 folds of their effective dose. PQ1 and PQ11 were also found to penetrate the blood brain barrier and collect in the tissue in significant amounts. The administration of PQ1 and PQ11 had no effect in the normal behavior of the animals indicating no short term adverse effects. PQ1 was found to increase the expression of survivin, an anti-apoptotic factor and decrease the expression of cleaved caspase-3 and caspase-8, pro-apoptotic proteins. These studies suggests that PQ1 might have anti-apoptotic activities in normal cells, in contrast to the role of PQ1 in cancer cells where it has demonstrated to induce apoptosis. The study also indicated that PQ11 was better metabolized from the tissues over time as compared to PQ1.