No description available.

Many insects and other arthropods have symbiotic microorganisms that may influence key facets of their biology. Cardinium hertigii is an intracellular bacterial symbiont, (phylum Bacteroidetes) of arthropods and nematodes. This versatile symbiont has been shown to cause three of four reproductive manipulations of their arthropod hosts known to be caused by symbionts: parthenogenesis induction (PI), where genetic males are converted into genetic females; feminization, where genetic males become functional females; and cytoplasmic incompatibility (CI), the symbiont-induced death of offspring from matings of infected males and uninfected females. Here, I explored the evolution of this symbiont and its reproductive manipulations, and found that closely related Cardinium strains have a tendency to associate with closely related hosts and the reproductive manipulations do not display a clear phylogenetic signal. To further understand the possible genes underlying these reproductive manipulations, I sequenced four Cardinium genomes and compared these with the two genomes analyzed in the literature. In these comparisons, I found that, although closely related Cardinium strains tend to reside in closely related hosts, there is no evidence for a suite of genes associated with host specificity, as few differences separate two strains residing in different host orders, suggesting that ecological opportunity for horizontal transmission may be more limiting to Cardinium than genomic capability. I additionally identify some genes that may be associated with the Cardinium’s ability to induce PI and CI in its wasp host. Overall, this dissertation has led to a better understanding of Cardinium and its effects on its hosts.

Wolbachia are maternally inherited, obligate, intracellular bacteria inducing a form of sterility known as cytoplasmic incompatibility. Wolbachia based strategies have been proposed for the control of disease vectors. One example is to use a population replacement strategy to drive into natural population a novel Wolbachia that modifies the age structure of a vector population, reducing disease transmission. In this research, the effects of a life-shortening stain of Wolbachia (popcorn Wolbachia) are transferred into the mosquitoes Aedes albopictus (Chapter Two and Three) and A. aegypti (Chapter Four and Five). In Chapter Two, the Wolbachia symbiosis significantly reduced fecundity and egg hatches in A. albopictus, with Wolbachia being highly pathogenic in this mosquito species. In Chapter Three, the relationship between popcorn Wolbachia and its host (in a triple-infected mosquito strain) varied with the mosquito diet. Feeding on mouse blood was associated with the loss of infection, whereas the infection was maintained in human blood-fed mosquito lines. Egg viability of triple infected mosquito was reduced only with mouse blood. In Chapter Four, the reduced competitiveness (e.g., low survival and increased developmental time) of infected A. aegypti immatures was associated with popcorn Wolbachia, relative to uninfected individuals in low food condition. In Chapter Five, the decreased survival of immature A. aegypti was associated with popcorn Wolbachia in the presence of potential predators (i.e., older A. aegypti or A. albopictus larvae). Using a novel behavioral assay, a delayed larval reaction to light avoidance was observed to be associated with the infection, suggesting Wolbachia effects on immature host behaviors. In Chapter Six, popcorn Wolbachia and wAlbB infected A. aegypti showed similar reproduction potential. No reduced level of CI or mating competitiveness was observed in wAlbB infected males. The results suggest the wAlbB infection in A. aegypti can be an additional agent for Wolbachia-based control strategies. In Chapter Seven, a filtering system using commercially available sieves was able to separate immature mosquitoes from water, preventing escape of mosquitoes. In Chapter Eight, an inexpensive artificial blood feeding was designed for feeding multiple mosquito cages. The results support the use of these tools to facilitate mass rearing of mosquitoes.

Wolbachia is a maternally inherited α-proteobacteria that lives an obligate intracellular lifestyle in its eukaryotic hosts. Wolbachia pipientis was first identified in the 1920’s in the mosquito Culex pipiens; Wolbachia has since been detected in a broad range of hosts. Wolbachia induces a number of diverse reproductive manipulations in many of its hosts, implicating Wolbachia in control strategies for target species. However, changes in host bacterial interactions can evolve toward mutualism, and this requires investigation in target systems. / This first part of this work examines changes in the Drosophila simulans-Wolbachia Riverside system in California. This system, studied for a period of more than two decades, provides a rare example in nature of a symbiotic relationship shifting from parasitism to mutualism. In my study, no changes in overall estimates of infection frequency, maternal transmission or cytoplasmic incompatibility were found, however there appears to be heterogeneity in maternal transmission rates and the ability of males to induce cytoplasmic incompatibility (CI). The source of CI variation in one isofemale line was subsequently investigated through reciprocal backcrossing with an old Wolbachia-infected strain, Riv88, and the variation was attributed to the Wolbachia genome. Further investigations are required to determine the extent of CI variation in the wild. These findings significant implications for the many applied projects that intend to exploit Wolbachia for its ability to manipulate host reproductive systems. / The second part of this work investigates the extent to which evolution of a host-bacterial symbiosis can influence the effectiveness of control strategies using Wolbachia. A particular strain of Wolbachia, named popcorn (wMelPop), has been isolated for its ability to reduce host longevity. This bacterium has the capacity to alter the age structure of the target host populations. As many vector-borne diseases (eg: Dengue Fever, Malaria) require an incubation period within the vector prior to transmission to the final host, a reduction in the lifespan of vector populations has the potential to lower transmission levels. The long term stability of this bacterial strain has been investigated in its native host (Drosophila melanogaster), and a novel host (D. simulans). The longevity-shortening phenotype and several life-history traits that might influence the effectiveness of control strategies have been examined. Host genetic background and the indirect selection on the longevity phenotype (via early or late fecundity selection) were found to influence multiple life-history traits in these species. The source of this variation in D. melanogaster is attributed primarily to the host, but can also be influenced by Wolbachia. Strong host background effects were noted in D. simulans, which highlights the importance of testing multiple backgrounds for suitability of use in applied projects. / This work has allowed for greater understanding of the potential for evolution within host-symbiont systems, and can be used to provide a framework for undertaking projects that will use Wolbachia as biological control agents. Future projects should consider the potential for evolution in detail under laboratory conditions before control strategies are implemented in the wild.

Wolbachia is a maternally inherited α-proteobacteria that lives an obligate intracellular lifestyle in its eukaryotic hosts. Wolbachia pipientis was first identified in the 1920’s in the mosquito Culex pipiens; Wolbachia has since been detected in a broad range of hosts. Wolbachia induces a number of diverse reproductive manipulations in many of its hosts, implicating Wolbachia in control strategies for target species. However, changes in host bacterial interactions can evolve toward mutualism, and this requires investigation in target systems. / This first part of this work examines changes in the Drosophila simulans-Wolbachia Riverside system in California. This system, studied for a period of more than two decades, provides a rare example in nature of a symbiotic relationship shifting from parasitism to mutualism. In my study, no changes in overall estimates of infection frequency, maternal transmission or cytoplasmic incompatibility were found, however there appears to be heterogeneity in maternal transmission rates and the ability of males to induce cytoplasmic incompatibility (CI). The source of CI variation in one isofemale line was subsequently investigated through reciprocal backcrossing with an old Wolbachia-infected strain, Riv88, and the variation was attributed to the Wolbachia genome. Further investigations are required to determine the extent of CI variation in the wild. These findings significant implications for the many applied projects that intend to exploit Wolbachia for its ability to manipulate host reproductive systems. / The second part of this work investigates the extent to which evolution of a host-bacterial symbiosis can influence the effectiveness of control strategies using Wolbachia. A particular strain of Wolbachia, named popcorn (wMelPop), has been isolated for its ability to reduce host longevity. This bacterium has the capacity to alter the age structure of the target host populations. As many vector-borne diseases (eg: Dengue Fever, Malaria) require an incubation period within the vector prior to transmission to the final host, a reduction in the lifespan of vector populations has the potential to lower transmission levels. The long term stability of this bacterial strain has been investigated in its native host (Drosophila melanogaster), and a novel host (D. simulans). The longevity-shortening phenotype and several life-history traits that might influence the effectiveness of control strategies have been examined. Host genetic background and the indirect selection on the longevity phenotype (via early or late fecundity selection) were found to influence multiple life-history traits in these species. The source of this variation in D. melanogaster is attributed primarily to the host, but can also be influenced by Wolbachia. Strong host background effects were noted in D. simulans, which highlights the importance of testing multiple backgrounds for suitability of use in applied projects. / This work has allowed for greater understanding of the potential for evolution within host-symbiont systems, and can be used to provide a framework for undertaking projects that will use Wolbachia as biological control agents. Future projects should consider the potential for evolution in detail under laboratory conditions before control strategies are implemented in the wild.

Wolbachia est une bactérie endosymbiotique, intracellulaire et exclusivement transmise maternellement qui infecterait au moins 106 espèces d'insectes. Wolbachia manipule fréquemment la reproduction de ses hôte à son avantage, notamment en induisant une forme de stérilité conditionnelle appelée incompatibilité cytoplasmique (IC). Chez les moustiques du complexe Culex pipiens, une grande diversité de souches de Wolbachia et de types d'IC a été précédemment identifiée, mais plusieurs aspects de la biologie de cette association restaient peu connus. Les travaux présentés dans cette thèse ont notamment permis de caractériser (i) l'impact de Wolbachia sur la structuration génétique des populations hôtes et (ii) la diversité des souches de Wolbachia et, plus précisément d'appréhender le mécanisme de l'IC. Par un suivi de populations naturelles, nous avons mis en évidence que Wolbachia induisait une forte structuration de la diversité mitochondriale, mais aussi qu'elle participait à des événements répétés d'introgression cytoplasmique entre les différents membres du complexe Cx. pipiens. Nous avons également mené une étude de génomique comparative basée sur le séquençage de quatre génomes complets de Wolbachia très proches phylogénétiquement. Pour cela, nous avons mis en place une série d'analyses approfondies utilisant un large panel d'outils bioinformatiques couplés à des vérifications moléculaires. Nous avons montré qu'il existait peu de polymorphisme entre les groupes de Wolbachia infectant Cx. pipiens. De plus, ces études nous ont permis de mettre en évidence des gènes candidats qui pourraient être directement impliqués dans le mécanisme de l'IC. / Wolbachia is an intracellular bacterial symbiont, exclusively maternally inherited, infecting at least 106 species of insects. Wolbachia commonly manipulates insect reproduction to its own advantage, as well illustrated by a phenomenon of conditional sterility called cytoplasmic incompatibility (CI). In mosquitoes of Culex pipiens complex, a great diversity of Wolbachia strains and of CI types was previously identified, but several aspects of the biology of this symbiotic association remained unknown. The aim of the studies presented in this thesis is to characterize (i) the impact of Wolbachia on the host genetic structure and (ii) the Wolbachia strains diversity in order to attempt an identification of CI molecular basis. By a survey of natural populations, we highlighted that Wolbachia deeply impacts the population structure of mitochondrial diversity, but is also associated with repeated events of cytoplasmic introgression between members of complex Cx. pipiens. We also conducted a study of comparative genomics based on the sequencing of four complete genomes of very closely related Wolbachia strains. For that purpose, we performed a series of analyses using a wide panel of bioinformatic tools coupled with molecular validations. We showed a low polymorphism between two groups of Wolbachia infecting Cx. pipiens. These studies also allowed us to highlight promising candidate genes which could be directly involved in the CI mechanism.

No description available.

Spiders are host to a plethora of heritable endosymbiotic bacteria. Broad-taxa screening studies indicate that endosymbionts are particularly common among spiders, however, little is known about how these bacteria affect their spider hosts. In insects these bacteria ensure vertical transmission by either conveying a benefit to the host or manipulating host reproduction to eliminate males that serve as evolutionary dead-ends for maternally-inherited bacteria. Common modes of reproductive manipulation include parthenogenesis, male killing, feminization, and cytoplasmic incompatibility. Screening an assemblage of Mermessus genus spiders, I detected a high frequency and diversity of endosymbiont infection. Within a single species, M. fradeorum, I detected three endosymbionts in multiple combinations. Rearing two natural infection types of M. fradeorum demonstrated two distinct endosymbiotic reproductive manipulations. Mothers infected with Rickettsia and Wolbachia produced extremely female-biased offspring, and antibiotic elimination of the symbionts successfully restored the sex ratio to the expected 1:1 in subsequent generations. A two-way factorial mating assay detected strong cytoplasmic incompatibility induced by a different strain of Wolbachia: cured females mated with infected males produced 70% fewer offspring than all other pairings. These results show that M. fradeorum is subject to multiple layers of reproductive manipulation that likely drive host evolution and ecology.

Cytoplasmic incompatibility (CI) is an intriguing, widespread, symbiont-induced reproductive failure that decreases offspring production of arthropods through crossing incompatibility of infected males with uninfected females or with females infected with a distinct symbiont genotype. For years, the molecular mechanism of CI remained unknown. Recent genomic, proteomic, biochemical, and cell biological studies have contributed to understanding of CI in the alphaproteobacterium Wolbachia and implicate genes associated with the WO prophage. Besides a recently discovered additional lineage of alphaproteobacterial symbionts only moderately related to Wolbachia, Cardinium (Bacteroidetes) is the only other symbiont known to cause CI, and genomic evidence suggests that it has very little homology with Wolbachia and evolved this phenotype independently. Here, we present the first transcriptomic study of the CI Cardinium strain cEper1, in its natural host, Encarsia suzannae, to detect important CI candidates and genes involved in the insect-Cardinium symbiosis. Highly expressed transcripts included genes involved in manipulating ubiquitination, apoptosis, and host DNA. Female-biased genes encoding ribosomal proteins suggest an increase in general translational activity of Cardinium in female wasps. The results confirm previous genomic analyses that indicated that Wolbachia and Cardinium utilize different genes to induce CI, and transcriptome patterns further highlight expression of some common pathways that these bacteria use to interact with the host and potentially cause this enigmatic and fundamental manipulation of host reproduction. IMPORTANCE The majority of insects carry maternally inherited intracellular bacteria that are important in their hosts' biology, ecology, and evolution. Some of these bacterial symbionts cause a reproductive failure known as cytoplasmic incompatibility (CI). In CI, the mating of symbiont-infected males and uninfected females produces few or no daughters. The CI symbiont then spreads and can have a significant impact on the insect host population. Cardinium, a bacterial endosymbiont of the parasitoid wasp Encarsia in the Bacteroidetes, is the only bacterial lineage known to cause CI outside the Alphaproteobacteria, where Wolbachia and another recently discovered CI symbiont reside. Here, we sought insight into the gene expression of a CI-inducing Cardinium strain in its natural host, Encarsia suzannae. Our study provides the first insights into the Cardinium transcriptome and provides support for the hypothesis that Wolbachia and Cardinium target similar host pathways with distinct and largely unrelated sets of genes.

Depuis sa récente invasion dans les continents européen et américain, la drosophile à aile tachetées, Drosophila suzukii est devenue un ravageur majeur des cultures de fruits rouges. Contrairement aux autres espèces de drosophiles, D. suzukii, est capable de pondre ses œufs dans des fruits sains avant la récolte, à l'aide de son ovipositeur sclérotinisé. Les pertes économiques liées à la présence de D. suzukii s'élèvent annuellement à plusieurs millions de dollars. Le contrôle des populations se fait principalement par l'utilisation de pesticides. Ici, nous avons testé si la bactérie Wolbachia pouvait être efficace pour lutter contre cette espèce. Ce symbiote est présent chez de nombreuses espèces d'insectes et induit souvent de l'incompatibilité cytoplasmique (IC) : les descendants des mâles infectés meurent, exceptés si l'œuf est sauvé par la même infection, héritée de la mère et qui va protéger l'embryon contre cette toxine encore non identifiée. La Technique de l'Insecte Incompatible (TII) repose sur l'utilisation de l'IC pour contrôler les populations d'insecte par des lâchers de mâles infectés. Nous avons montré que D. suzukii est naturellement infecté par une souche de Wolbachia, nommée wSuz, avec des prévalences intermédiaire et qui n'induit pas un taux d'IC élevé. Pour le développement de la TII chez D. suzukii, nous avons réalisé des transferts de souches de Wolbachia entre D. simulans et D. suzukii pour identifier des souches qui peuvent stériliser les femelles D. suzukii, en dépit de la présence de wSuz. Nous avons identifié deux souches de Wolbachia comme candidates pour le développement de la TII. Ces souches induisent des taux d'IC très élevés chez ce ravageur, qui n'est pas atténué par la présence de wSuz chez les femelles. Les mâles stérilisants ont une compétitivité sexuelle similaire comparés à celle des mâles infectés ou non par wSuz, et sont capable d'induire des taux d'IC élevés tout au long de leurs vie. Finalement nous avons montré que, dans de grandes cages à population, la TII pouvait être très efficace pour limiter l'augmentation de la taille des populations de D. suzukii. L'ensemble des résultats confirment que la TII est une approche prometteuse pour contrôler les populations de D. suzukii et mérite de dépasser le stade du laboratoire. Associé à une technique de sexage efficace, la TII peut être un outil puissant, spécifique et respectueux de l'environnement / Since its recent invasion of the European and American continents, the spotted wing Drosophila, D. suzukii has become a major burden of the fruit industry. Armed with a highly sclerotized ovipositor, females can lay eggs in a wide variety of ripening and healthy fruits, in contrast to other Drosophila species. Economic losses due to D. suzukii reach millions of dollars annually and methods to control natural populations in the field mainly rely on the use of chemical pesticides. Here we test if Wolbachia bacteria can represent a potential ally to control this pest species. These symbionts are naturally present in many insects and often induce a form of conditional sterility called Cytoplasmic Incompatibility (CI): the offspring of infected males die, unless the eggs are rescued by the same infection inherited from the mother which protects the embryo against a yet unidentified toxin. As long recognized, a strategy called the Incompatible Insect Technique (IIT) makes use of the CI phenotype to control insect populations through the mass release of infected males. D. suzukii is naturally infected by a single Wolbachia strain, named wSuz, which has an intermediate prevalence in field populations and which does not induce a high level of CI. To implement IIT in D. suzukii, we used back and forth Wolbachia transfers between D. suzukii and D. simulans to identify Wolbachia strains that can fully sterilize D. suzukii females despite the presence of wSuz. We identified two potential candidates, both induce a very high level of CI in this pest which is not attenuated by the presence of wSuz in females. The transinfected males showed a similar competitiveness compared naturally infected and uninfected males and are able to induce a high level of CI during all their life. Finally we demonstrated that, in large population cage, the IIT can be very efficient to limit the D. suzukii population size. All the results confirmed that the IIT is a promising approach to control D. suzukii population and merit to go out the laboratory. Associate with a perfect sexing technique, IIT can be a powerful tool to fight against D. suzukii, which is not polluting and species specific