Aedes aegypti mosquitoes infected with Wolbachia bacteria are currently being released for arbovirus suppression around the world. Their potential to invade populations and persist will depend on interactions with environmental conditions, particularly as larvae are often exposed to fluctuating and extreme temperatures in the field. We reared Ae. aegypti larvae infected with different types of Wolbachia (wMel, wAlbB and wMelPop-CLA) under diurnal cyclical temperatures. Rearing wMel and wMelPop-CLA-infected larvae at 26–37°C reduced the expression of cytoplasmic incompatibility, a reproductive manipulation induced by Wolbachia. We also observed a sharp reduction in the density of Wolbachia in adults. Furthermore, the wMel and wMelPop-CLA infections were not transmitted to the next generation when mosquitoes were exposed to 26–37°C across all life stages. In contrast, the wAlbB infection was maintained at a high density, exhibited complete cytoplasmic incompatibility, and was transmitted from mother to offspring with a high fidelity under this temperature cycle. These findings have implications for the potential success of Wolbachia interventions across different environments and highlight the importance of temperature control in rearing. There is currently great interest in using the bacterium Wolbachia to reduce the burden of dengue and Zika; viruses which infect millions of people globally each year. Aedes aegypti mosquitoes with Wolbachia infections can invade natural populations and interfere with the transmission of these viruses. However, we find that the wMel strain of Wolbachia which is currently being used for dengue and Zika control in several countries may have reduced effectiveness at invading populations when mosquitoes experience heat stress. Since mosquito larvae experience extreme temperatures in their natural habitat, our results have implications for current and future releases of Wolbachia-infected mosquitoes and highlight the need for further investigation into alternative strains of Wolbachia.

Wolbachiaare endosymbionts found in many insects with the potential to suppress vectorborne diseases, particularly through interfering with pathogen transmission.Wolbachiastrains are highly variable in their effects on hosts, raising the issue of which attributes should be selected to ensure that the best strains are developed for disease control. This depends on their ability to suppress viral transmission, invade host populations, persist without loss of viral suppression and not interfere with other control strategies. The potential to achieve these objectives is likely to involve evolutionary constraints; viral suppression may be limited by the ability of infections to spread due to deleterious host fitness effects. However, there are exceptions to these patterns in both natural infections and in novel associations generated following interspecific transfer, suggesting that pathogen blockage, deleterious fitness effects and changes to reproductive biology might be at least partly decoupled to achieve ideal infection attributes. The stability of introducedWolbachiaand its effects on viral transmission remain unclear, but rapid evolutionary changes seem unlikely. Although deliberate transfers ofWolbachiaacross species remain particularly challenging, the availability of strains with desirable attributes should be expanded, taking advantage of the diversity available across thousands of strains in natural populations.

Wolbachia is an endosymbiotic Alphaproteobacteria that can suppress insect-borne diseases through decreasing host virus transmission (population replacement) or through decreasing host population density (population suppression). We contrast natural Wolbachia infections in insect populations with Wolbachia transinfections in mosquitoes to gain insights into factors potentially affecting the long-term success of Wolbachia releases. Natural Wolbachia infections can spread rapidly, whereas the slow spread of transinfections is governed by deleterious effects on host fitness and demographic factors. Cytoplasmic incompatibility (CI) generated by Wolbachia is central to both population replacement and suppression programs, but CI in nature can be variable and evolve, as can Wolbachia fitness effects and virus blocking. Wolbachia spread is also influenced by environmental factors that decrease Wolbachia titer and reduce maternal Wolbachia transmission frequency. More information is needed on the interactions between Wolbachia and host nuclear/mitochondrial genomes, the interaction between invasion success and local ecological factors, and the long-term stability of Wolbachia-mediated virus blocking.

Insects and other invertebrates can readily adapt to a range of environmental conditions and these include conditions used in artificial rearing. This can lead to problems when mass rearing insects and mites for release as biocontrol agents or in sterile insect control programs, and when using laboratory strains to understand field population dynamics. Laboratory adaptation experiments also help to understand potential rates of trait evolution and repeatability of evolutionary changes. Here, we review evidence for laboratory adaptation across invertebrates, contrasting different taxonomic groups and providing estimates of the rate of evolutionary change across trait classes. These estimates highlight rapid changes in the order of 0.033 (median) haldanes and up to 2.4 haldanes, along with proportional changes in traits of more than 10% per generation in some cases. Traits tended to change in the direction of increased fitness for Coleoptera, Diptera and Hymenoptera, but changes in Lepidoptera were often in the opposite direction. Laboratory-adapted lines tend to be more sensitive to stress, likely reflecting relaxed selection for stress-related traits. Morphological traits show smaller changes under laboratory conditions than other types of traits. Estimates of evolutionary rates slowed as more generations were included in comparisons, perhaps reflecting nonlinear dynamics although such patterns may also reflect variance differences among trait classes. The rapid rate of laboratory adaptation in some cultures reinforces the need to develop guidelines for maintaining quality during mass rearing and highlights the need for caution when using laboratory lines to represent the performance of species in vulnerability assessments.

Wolbachia bacteria are now being introduced into Aedes aegypti mosquito populations for dengue control. When Wolbachia infections are at a high frequency, they influence the local transmission of dengue by direct virus blocking as well as deleterious effects on vector mosquito populations. However, the effectiveness of this strategy could be influenced by environmental temperatures that decrease Wolbachia density, thereby reducing the ability of Wolbachia to invade and persist in the population and block viruses. We reared wMel-infected Ae. aegypti larvae in the field during the wet season in Cairns, North Queensland. Containers placed in the shade produced mosquitoes with a high Wolbachia density and little impact on cytoplasmic incompatibility. However, in 50% shade where temperatures reached 39°C during the day, wMel-infected males partially lost their ability to induce cytoplasmic incompatibility and females had greatly reduced egg hatch when crossed to infected males. In a second experiment under somewhat hotter conditions (>40°C in 50% shade), field-reared wMel-infected females had their egg hatch reduced to 25% when crossed to field-reared wMel-infected males. Wolbachia density was reduced in 50% shade for both sexes in both experiments, with some mosquitoes cleared of their Wolbachia infections entirely. To investigate the critical temperature range for the loss of Wolbachia infections, we held Ae. aegypti eggs in thermocyclers for one week at a range of cyclical temperatures. Adult wMel density declined when eggs were held at 26–36°C or above with complete loss at 30–40°C, while the density of wAlbB remained high until temperatures were lethal. These findings suggest that high temperature effects on Wolbachia are potentially substantial when breeding containers are exposed to partial sunlight but not shade. Heat stress could reduce the ability of Wolbachia infections to invade mosquito populations in some locations and may compromise the ability of Wolbachia to block virus transmission in the field. Temperature effects may also have an ecological impact on mosquito populations given that a proportion of the population becomes self-incompatible. Aedes aegypti mosquitoes infected with Wolbachia symbionts are being deployed in the tropics as a way of reducing disease transmission. Some Wolbachia strains are vulnerable to high temperatures but these effects have not been evaluated outside of a laboratory setting. We reared Ae. aegypti infected with the wMel strain of Wolbachia in the field during the wet season in Cairns, Australia, where the first releases of Wolbachia-infected Ae. aegypti took place. wMel-infected mosquitoes became partially self-incompatible, with reduced egg hatch, when larvae were reared in partial shade where maximum daily temperatures exceeded 39°C. Under these conditions the amount of Wolbachia in adult mosquitoes was reduced to less than 1% of laboratory-reared mosquitoes on average, while some mosquitoes were cleared of Wolbachia entirely. In contrast, wMel was stable when mosquitoes were reared under cooler conditions in full shade. Field trials with the wMel strain are now underway in over 10 countries, but high temperatures in some locales may constrain the ability of Wolbachia to invade natural mosquito populations and block disease transmission.

Wolbachia endosymbionts are potentially useful tools for suppressing disease transmission by Aedes aegypti mosquitoes because Wolbachia can interfere with the transmission of dengue and other viruses as well as causing deleterious effects on their mosquito hosts. Most recent research has focused on the wMel infection, but other infections also influence viral transmission and may spread in natural populations. Here, we focus on the wAlbB infection in an Australian outbred background and show that this infection has many features that facilitate its invasion into natural populations including strong cytoplasmic incompatibility, a lack of effect on larval development, an equivalent mating success to uninfected males and perfect maternal transmission fidelity. On the other hand, the infection has deleterious effects when eggs are held in a dried state, falling between wMel and the more virulent wMelPop Wolbachia strains. The impact of this infection on lifespan also appears to be intermediate, consistent with the observation that this infection has a titer in adults between wMel and wMelPop. Population cage experiments indicate that the wAlbB infection establishes in cages when introduced at a frequency of 22%, suggesting that this strain could be successfully introduced into populations and subsequently persist and spread.