Biology subjects

Ross, P. A.

Publications and source records attributed to Ross, P. A..

At least 19 recordsLinked to original sources

bioRxiv2026

Efficacy of 60 personal mosquito repellents and perfumes against Aedes aegypti mosquitoes

Personal insect repellents can offer an effective means of preventing mosquito bites. Despite a wide variety of products that claim to repel mosquitoes, relatively few studies have systematically compared their performance through standardized tests. In Australia, chemical-based repellents must be registered with the Australian Pesticides and Veterinary Medicines Authority (APVMA); however, many unregistered products with unverified effectiveness remain commercially available. Here, we tested 55 insect repellents and 5 non-repellent perfumes and deodorants sold in Australia for their protection against Aedes aegypti mosquito landings (as a proxy for bites) using arm-in-cage assays across eight human subjects. We tested 43 topical repellents (containing 18 different primary active ingredients), four wristbands, three clothing stickers, three ultrasonic devices, one garment, one repellent applied to fabric, two perfumes, two deodorants and one antiperspirant. Thirteen repellents, including all wristbands, stickers and ultrasonic devices did not provide complete protection against mosquito landings for any human subject, while a further sixteen provided complete protection for 1 hr or less on average. Repellents containing DEET, oil of lemon eucalyptus, picaridin and IR3535 as the primary active ingredient were the only products that provided more than 2 hours of complete protection, but with significant variation among products with the same active ingredient. Only 7/30 essential oil-based topical repellents and perfumes provided complete protection for greater than 1 hr. Nearly half (25/52) of the repellents subject to registration with the APVMA were unregistered despite being advertised or labelled as mosquito repellents, and these tended to be less effective than registered products. Only 5/28 repellents met or exceeded their claimed protection time. This study highlights the prevalence of unregistered and ineffective mosquito repellents on the Australian market while providing evidence for products and active ingredients that do protect against mosquito bites under laboratory conditions.

zoology↗

bioRxiv2026

Mosquito fertility is (mostly) resilient to sublethal heat shocks

Insects can experience a loss of fertility under acute heat stress, but impacts vary among species and depend on life stage and sex. We examined the effects of sublethal heat shocks on the fertility of three mosquito species (Aedes aegypti, Aedes notoscriptus and Culex quinquefasciatus) when males or females were exposed at the pupal and adult stages. Reproductive performance was assessed through egg-laying success, fecundity, egg hatchability and viable offspring production. Female heat shock had limited effects on fertility in all three species, though exposure of adult females to 41 {degrees}C for 1 hr reduced viable offspring production in Ae. notoscriptus. Male heat shock reduced egg hatchability in Cx. quinquefasciatus and viable offspring production in Ae. aegypti depending on the life stage exposed. We investigated several other aspects of heat shock exposure on Ae. aegypti fertility. In this species, male fertility partially recovered within 24-48 hours following heat shock exposure, while effects of heat shocks on female fertility occurred regardless of whether heat exposure occurred pre- or post-mating. In addition, maternal heat shock exposure enhanced offspring fecundity and viable offspring production under subsequent sublethal heat stress. Overall, our findings demonstrate that mosquito fertility is largely robust to short-term heat shocks, but there are also complex trait, sex, stage, temporal and species-specific effects, with implications for predicting mosquito population persistence and species persistence under future climate extremes.

zoology↗

bioRxiv2026

Interactions between inbreeding, fitness and the bacterial microbiome in Aedes aegypti mosquitoes

Laboratory and field populations of insects can experience a decline in fitness and loss of genetic diversity due to inbreeding depression and genetic drift, respectively. Matings among related individuals and small population size may also influence insect host microbiomes with consequences for fitness. In the dengue vector mosquito, Aedes aegypti, the bacterial microbiome is largely environmentally determined but recent studies have also revealed host genetic components. We generated a panel of 55 inbred lines from either of two founding outbred populations of Ae. aegypti to test for associations between life history traits, inbreeding, allelic diversity and microbiome composition using ddRADseq and bacterial 16S rRNA gene sequencing on pools of mosquitoes. Effects of inbreeding were diverse with severe composite fitness costs in many lines but minimal costs in others despite similar low levels of genetic diversity. We found no strong relationship between major life history traits across inbred lines, suggesting that any costs due to inbreeding were trait specific. Bacterial microbiome analysis of pooled samples from a subset of populations revealed common microbes across populations, with Elizabethkingia, Aeromonas and Ralstonia being the most abundant. Despite bacterial composition varying widely, there was no clear relationship between microbiome composition and fitness or population origin. However, there were several significant positive correlations between the relative abundance of different microbial taxa across lines. Our results demonstrate diverse impacts of inbreeding on fitness of mosquito populations but with limited impacts on the microbiome.

microbiology↗

bioRxiv2025

Heterogeneity in inhibition of genetically diverse dengue virus strains by Wolbachia

The release of Aedes aegypti mosquitoes transinfected with the virus-inhibiting Wolbachia bacterium has the potential to reduce the burden caused by dengue virus (DENV). However, the robustness of this control strategy across the wide genetic diversity of DENV remains unknown. Here, we systematically tested two commonly used Wolbachia strains (wAlbB and wMelM) for their ability to inhibit 60 genetically diverse DENV isolates spanning all four serotypes. We found stronger inhibition by wMelM (median relative dissemination: 0.04) than wAlbB (median relative dissemination: 0.19). Furthermore, while we found substantial heterogeneity in inhibition across DENV isolates, we found that more DENV-3 isolates were weakly inhibited (median relative dissemination: 0.47 for wAlbB and 0.39 for wMelM) compared to the other serotypes (median relative disseminations: 0.10-0.18 for wAlbB and 0-0.11 for wMelM). Using transmission dynamic models, we further showed that differential Wolbachia inhibition results in increased probability of reemergence, particularly in high transmission intensity settings, with strong selection for DENV strains that have higher relative dissemination in mosquitoes. Our work highlights the importance of considering DENV genetic diversity, including the long-term risk of selection, in Wolbachia-based control interventions.

microbiology↗

bioRxiv2025

Heat hardening enhances mosquito heat tolerance in a species-specific and trait-specific manner

Models predict that the distribution of ectotherms including mosquitoes will shift with climate change, but few incorporate adaptive capacity. Acclimation is one mechanism by which mosquitoes could adapt, allowing mosquitoes that have experienced sub-lethal stress previously to tolerate subsequent stressful environments. In this study we evaluated the heat tolerance of three vector mosquito species, Aedes aegypti, Ae. notoscriptus and Culex quinquefasciatus, after being previously exposed to heat hardening. Adult males and females were heat-hardened by exposure to 41{degrees}C for one hour and subsequently tested for heat survival and knockdown following one-hour heat shocks across a range of temperatures up to the lethal limit, ramping CTmax assays and static temperature knockdown time assays. The three species differed markedly in their heat tolerance across all assays, with Ae. aegypti being the most heat tolerant and Cx. quinquefasciatus being the least. Females from all three species were more heat tolerant than males in the one-hour heat shock assays, but effects of sex were absent or inconsistent for CTmax and heat knockdown time assays. A beneficial impact of heat hardening on subsequent heat shock knockdown was evident in both sexes of all three species. However, hardening effects differed substantially for survival 24 hr later, ranging from no effect of hardening in Cx. quinquefasciatus to a [~]1{degrees}C increase in LT50 in Ae. notoscriptus. In contrast, no effects of heat hardening were detected for CTmax or static knockdown time assays. An additional experiment in Ae. aegypti detected no benefits of heat shock exposure in female patents on the thermal tolerance of offspring. Our findings emphasize the need to consider effects of acclimation including heat hardening in models to predict the response of mosquitoes to climate warming. They also have implications for measuring thermal tolerance in mosquitoes more generally, given that both sex and hardening effects depend on the type of assay used and trait measured.

ecology↗

bioRxiv2025

Spiroplasma and heat hardening can buffer insect male fertility loss at high temperatures

Insects upper thermal limits for survival, activity, and fertility have been used to assess vulnerability to climate change, yet heritable endosymbionts - present in over 70% of insect species - are often overlooked. While emerging research suggests some endosymbionts can increase thermal tolerance, their effects on upper lethal and fertility thermal limits has rarely been investigated. Additionally, short-term exposure to sub-lethal high temperatures (heat hardening) can increase insect heat tolerance, but its impact on male fertility is unclear and potential interactions with endosymbionts has not been explored. Here, we investigate whether the endosymbiont Spiroplasma poulsonii and heat hardening influence upper survival and fertility thermal limits in a native widespread host of Spiroplasma (Drosophila hydei) and a novel host (Drosophila birchii), a rainforest-restricted species with low heat tolerance. Heat hardening generally improved survival and fertility following a heat shock. The presence of Spiroplasma increased survival and fertility of D. hydei males following heat-shock, while in D. birchii, it did not enhance survival but protected male fertility after heat-shock and modulated hardening responses at sub-lethal temperatures. While protective effects varied across species, sex, and trait, both Spiroplasma and hardening significantly buffered male fitness loss during heat shock in both host species and halved heat exposure risk under current and predicted climate change in the rainforest restricted D. birchii. These findings highlight that, beyond generating novel phenotypic variation, endosymbionts can interact with plastic responses to heat stress, emphasising the importance of accounting for endosymbiont-mediated effects on thermal tolerance when assessing insect vulnerability to climate change and exploring strategies to manipulate insect thermal tolerance.

evolutionary biology↗

bioRxiv2025

Specificities of Chemosensory Receptors in the Human Gut Microbiota

The human gut is rich in metabolites and harbors a complex microbial community, yet the sensory repertoire of its commensal bacteria remains largely uncharacterized. Here we systematically mapped ligand specificities of extracytoplasmic sensory domains from twenty members of the human gut microbiota, with a primary focus on the abundant and physiologically important class of Clostridia. We identified diverse metabolites as specific stimuli for three major functional classes of transmembrane receptors. We further characterized novel subsets of sensors belonging to the Cache superfamily, specific for lactate, dicarboxylic acids, and for uracil and short-chain fatty acids (SCFAs), respectively, and investigated the evolution of their ligand specificity. Structural and biochemical analysis of the newly described dCache_1UR domain revealed an independent binding of uracil and SCFA at distinct modules. Altogether, we could identify or predict specificities for over a half of the Cache-type chemotactic sensors in the selected gut commensals, with the carboxylic acids representing the largest class of ligands. Among those, the most commonly found specificities were for lactate and formate, indicating particular importance of these metabolites in the human gut microbiome and consistent with their observed beneficial impact on the growth of selected bacterial species.

microbiology↗

bioRxiv2025

Implications of successive blood feeding on Wolbachia-mediated dengue virus inhibition in Aedes aegypti mosquitoes

Dengue virus (DENV) is a mosquito-borne virus that poses a continued and increasing threat to public health. A promising strategy to mitigate the burden of DENV is introgression of the virus-inhibiting Wolbachia pipientis bacterium into Aedes aegypti populations in the field. While previous studies on Wolbachia-mediated virus inhibition have typically assessed viral replication following a single bloodmeal, the main vector of DENV, Ae. aegypti, feeds frequently, often biting multiple hosts per gonotrophic cycle and promptly attempting to refeed following egg laying. Previously, we demonstrated that successive blood feeding reduces the extrinsic incubation period (EIP) and shortens the time it takes for a mosquito to be able to transmit viruses to a new host. With this in mind, we investigated the impact of successive blood meals on DENV serotype 2 (DENV-2) in Ae. aegypti in the presence or absence of Wolbachia (wAlbB and wMelM). We found that both WT and Wolbachia transinfected had increased DENV-2 dissemination 7 days post-infection as well as higher body titers of DENV-2 in the double-fed groups. Using these empirical data in a binomial regression model, we estimated that successive feeding increased the probability of WT and Wolbachia transinfected mosquitoes surviving the EIP. When we estimated the odds of surviving the EIP for mosquitoes with Wolbachia relative to WT mosquitoes, successive feeding increased the chances of WT mosquitoes surviving the EIP more than in mosquitoes with Wolbachia, indicating a strong inhibitory effect of Wolbachia even in the context of natural frequent blood feeding behavior. Our work shows that mosquito feeding behavior should be considered when assessing the inhibitory effects of Wolbachia on DENV.

microbiology↗

bioRxiv2024

Elevated developmental temperatures below the lethal limit reduce Aedes aegypti fertility

Aedes aegypti mosquitoes are the principal vectors of dengue and continue to pose a threat to human health, with ongoing urbanization, climate change, and trade all impacting the distribution and abundance of this species. Hot periods are becoming increasingly common and their impacts on insect mortality have been well established, but they may have even greater impacts on insect fertility. In this study, we investigated the impacts of high temperatures on Ae. aegypti fertility both within and across generations. Mosquitoes developing under elevated temperatures exhibited higher critical thermal maxima (CTmax) reflecting developmental acclimation, but their fertility declined with increasing developmental temperature. In females, elevated developmental temperatures decreased fecundity while in males it tended to decrease egg hatch proportions and the proportion of individuals producing viable offspring. Rearing both sexes at 35{degrees}C increased fecundity in the subsequent generation but effects of elevated temperatures persisted across gonotrophic cycles within the same generation. Moreover, exposure of adults to 35{degrees}C further decreased fertility beyond the effects of developmental temperature alone. These findings highlight sub-lethal impacts of elevated temperatures on Ae. aegypti fertility and plastic responses to thermal stress within and across generations. This has significant implications for mosquito populations thriving in increasingly warmer environments.

ecology↗

bioRxiv2024

Endosymbionts impact ladybird predation rates of aphids in a temperature-dependent manner

Aphids are worldwide pests causing major economic losses to growers. Current management strategies rely heavily on pesticides, but some effective pesticides are being withdrawn and the efficiency of remaining pesticides is also decreasing as aphids build up resistance. Biological control using predators can provide a sustainable alternative to pesticides under some circumstances, while the deliberate introduction of facultative bacterial endosymbionts that induce host fitness costs and reduce plant virus transmission provides another potential future strategy to combat aphid pests. However, new control options should not be antagonistic, with the concern that the effectiveness of biocontrol might be altered by endosymbiont presence in hosts. We, therefore, tested if predation by two aphidophagous ladybirds, Adalia bipunctata and Harmonia conformis, on the green peach aphid, Myzus persicae, and the oat aphid, Rhopalosiphum padi, was affected by transinfected Rickettsiella viridis and both native and transinfected Regiella insecticola endosymbionts at different temperatures. The predation rate of aphids infected by either endosymbiont was higher at 14 {degrees} C than the rate for uninfected aphids of both species, but the opposite pattern was apparent at 20 {degrees}C and for one host-endosymbiont combination at 26 {degrees}C. Overall, the results showed that higher temperatures increased predation, while differences between intermediate and low temperatures were species-dependent. No transmission of endosymbionts from aphid to ladybird through predation was detected. These findings point to a lack of consistent effects of the investigated endosymbionts on predation rates in these major aphid pests. The temperature dependence of endosymbiont-predation interactions suggests that the impact of seasonal climate should be considered when assessing the potential of endosymbionts in a biological control setting. HIGHLIGHTS- Aphid endosymbiont effects on ladybird predation rates are temperature-dependent - Ladybird predation rate of aphids increases as temperature rises - Prey (aphid) endosymbionts are not transferred to predators (ladybirds)

ecology↗

bioRxiv2024

The endosymbiont Rickettsiella viridis increases the virulence of Diuraphis noxia but reduces alate frequency

Aphids are among the worlds most economically damaging pests and carry a diverse range of bacterial endosymbionts. There is increasing interest in exploring potential applications of natural and novel strains of endosymbionts in aphid control. One endosymbiont, Rickettsiella viridis, has a large fitness cost following transfer from its natural aphid host Acyrthosiphon pisum into a novel host aphid Myzus persicae. Here, we investigated host impacts after transferring the same Rickettsiella strain to an important cereal aphid, the Russian wheat aphid Diuraphis noxia. Rickettsiella in this host resulted in modest fitness effects, with a minor increase in heat tolerance and a decrease in development time at 25{degrees} C. The infection persisted in mixed caged populations under different temperatures. Surprisingly, Rickettsiella increased aphid virulence to wheat plants and to two non-crop hosts of D. noxia, barley grass and brome grass. This was evident from sharper decreases in leaf number and leaf area, as well as an increase in chlorotic streaking when plants were exposed to Rickettsiella-infected D. noxia. Rickettsiella also reduced the proportion of alates in aphids held in small cages and in larger mesocosms containing multiple wheat plants where short-distance dispersal of aphids was impacted. These results provide compelling evidence that Rickettsiella can affect virulence - the first case of an endosymbiont transfer directly influencing aphid virulence to host plants - and highlight the species-specific impacts of endosymbiont transfers on aphids which can involve multiple traits.

microbiology↗

bioRxiv2024

Wolbachia strain wMelM disrupts egg retention by Aedes aegypti females prevented from ovipositing

Aedes aegypti mosquitoes are well adapted to dry climates and can retain their eggs for extended periods in the absence of suitable habitat. Wolbachia strains transferred from other insects to mosquitoes can be released to combat dengue transmission by blocking virus replication and spreading through populations, but host fitness costs imposed by Wolbachia, particularly under some environments, can impede spread. We therefore assessed the impact of two Wolbachia strains being released for dengue control (wAlbB and wMelM) on fecundity and egg viability following extended egg retention (12 or 18 d) under laboratory conditions. Egg viability decreased to a greater extent in females carrying wMelM compared to uninfected or wAlbB females. Fertility fully recovered in uninfected females following a second blood meal after laying retained eggs, while wMelM females experienced only partial recovery. Effects of wMelM on egg retention were similar regardless of whether females were crossed to uninfected or wMelM males, suggesting that fitness costs were triggered by Wolbachia presence in females. The fecundity and hatch proportions of eggs of wMelM females declined with age, regardless of whether females used stored sperm or were recently inseminated. Costs of some Wolbachia strains during egg retention may affect the invasion and persistence of Wolbachia in release sites where larval habitats are scarce and/or intermittent.

ecology↗

bioRxiv2024

Hormetic effect induced by Beauveria bassiana in Myzus persicae

Myzus persicae, a serious sap-sucking pest of a large variety of host plants in agriculture, is traditionally controlled using chemical insecticides but there is interest in using biopesticides as restrictions are increasingly placed on the use of broad-spectrum pesticides. Here we show that in petri dish experiments high concentrations of the fungal entomopathogen Beauveria bassiana (strain PRRI 5336) lead to rapid mortality of M. persicae but at a low concentration (1 x 104 conidia mL-1) there is a hormetic effect where longevity and fecundity are enhanced. Hormetic effects persisted across a generation with reduced development times and increased fecundity in the offspring of M. persicae exposed to B. bassiana. Whole plant experiments point to a hormetic effect being detected in two out of three tested lines. The impact of these effects might also depend on whether M. persicae was transinfected with the endosymbiont Rickettsiella viridis, which decreases fecundity and survival compared to aphids lacking this endosymbiont. This fecundity cost was ameliorated in the generation following exposure to the entomopathogen. While B. bassiana is effective in controlling M. persicae especially at higher spore concentrations, utilization of this entomopathogen requires careful consideration of hormetic effects at lower spore concentrations, and further research to optimize its application for sustainable agriculture is recommended. AUTHOR SUMMARYBiopesticides such as Beauveria bassiana can be effective alternatives to chemical insecticides to control insect pests. We tested the efficacy of this biopesticide against the important agricultural pest aphid Myzus persicae in laboratory experiments. We also tested whether the potential biological control agent and endosymbiont Rickettsiella viridis could provide protection against mortality caused by B. bassiana. While high doses of B. bassiana caused rapid mortality in aphids, low doses enhanced aphid fecundity and survival. This enhancement persisted into the next generation, with shortened development times and increased fecundity regardless, even when high doses were used in the previous generation. The endosymbiont R. viridis did not provide clear protection against B. bassiana, in contrast to previous studies in other aphid species, but beneficial effects at low doses also occurred in this aphid line. We also observed hormesis on experiments on whole plants, but only for some aphid genotypes. To a lesser extent, we also observed beneficial effects of low doses of B. bassiana in experiments on whole plants, but only in some aphid genotypes. Fitness enhancement by biopesticides at low doses raises concerns for field applications but further research is required to understand its underlying mechanisms.

ecology↗

bioRxiv2023

Wolbachia infection negatively impacts Drosophila simulans heat tolerance in a strain- and trait-specific manner

The susceptibility of insects to rising temperatures has largely been measured by their ability to survive thermal extremes. However, until recently, the capacity for maternally inherited endosymbionts to influence insect heat tolerance has been overlooked. Further, the impact of heat on traits like fertility, which can decline at temperatures below the lethal thermal limit has largely been ignored. Here, we assess the impact of three Wolbachia strains (wRi, wAu, and wNo) on the survival and fertility of Drosophila simulans exposed to heat stress during development or as adults. The impact of Wolbachia infection on heat tolerance was generally small and trait/strain specific. Only the wNo infection significantly reduced survival and fertility of adult males after a heat shock. When exposed to a fluctuating heat stress during development, the wRi and wAu strains reduced egg-to-adult survival but only the wNo infection reduced male fertility. Wolbachia densities of all three strains decreased under developmental heat stress, but reductions occurred at temperatures above those that reduced fertility of the host. These findings reveal the complexity of endosymbiont-host-environment interactions and emphasise the necessity to account for endosymbionts and their effect on both survival and fertility when investigating the vulnerability of insects to climate change.

evolutionary biology↗

bioRxiv2023

CD22L Conjugation to Insulin Attenuates Insulin-Specific B cell Activation

Pancreatic islet-reactive B lymphocytes promote Type 1 diabetes (T1D) by presenting antigen to islet-destructive T cells. Teplizumab, an anti-CD3 monoclonal, delays T1D onset in patients at risk, but additional therapies are needed to prevent disease entirely. Therefore, bifunctional molecules were designed to selectively inhibit T1D-promoting anti-insulin B cells by conjugating a ligand for the B cell inhibitory receptor CD22 (i.e., CD22L) to insulin, which permit these molecules to concomitantly bind to anti-insulin B cell receptors (BCRs) and CD22. Two prototypes were synthesized: 2:2 insulin-CD22L conjugate on a 4-arm PEG backbone, and 1:1 insulin-CD22L direct conjugate. Transgenic mice (125TgSD) expressing anti-insulin BCRs provided cells for in vitro testing. Cells were cultured with constructs for three days then assessed by flow cytometry. Duplicate wells with anti-CD40 simulated T cell help. Surprisingly, a 2-insulin 4-arm PEG control caused robust proliferation and activation-induced CD86 upregulation. Anti-CD40 further boosted these effects. This was unexpected, as soluble insulin alone has no effect, and may indicate that BCR-crosslinking occurs when antigens are tethered by the PEG backbone. Addition of CD22L via the 2:2 insulin-CD22L conjugate restored B cell properties to that of controls without additional beneficial effect. In contrast, the 1:1 insulin-CD22L direct conjugate significantly reduced anti-insulin B cell proliferation in the presence of anti-CD40. CD22L alone had no effect, and the constructs did not affect WT B cells. Thus, high valency constructs activate anti-insulin B cells, while low-valency antigen-CD22L conjugates co-ligate BCR and CD22, reducing B cell activation in response to simulated T cell help and reducing pathogenic B cell numbers without harming normal cells. Thus, the insulin-CD22L direct conjugate is a promising candidate for preclinical trials to prevent T1D without inducing immunodeficiency

bioengineering↗

bioRxiv2023

How often are male mosquitoes attracted to humans?

Many mosquito species live close to humans where females feed on human blood. While male mosquitoes do not feed on blood, it has long been recognized that males of some species can be attracted to human hosts. To investigate the frequency of male mosquito attraction to humans, we conducted a literature review and human-baited field trials, as well as laboratory experiments involving males and females of three common Aedes species. Our literature review indicated that male attraction to humans is limited to a small number of species, including Ae. aegypti and Ae. albopictus. In our human-baited field collections, only 4 out of 13 species captured included males. In laboratory experiments, we found that male Ae. notoscriptus and Ae. vigilax showed no attraction to humans, while male Ae. aegypti exhibited persistent attraction for up to 30 minutes. Both male and female Ae. aegypti displayed similar preferences for different human subjects, suggesting that male Ae. aegypti respond to similar cues as females. Additionally, we found that mosquito repellents applied to human skin effectively repelled male mosquitoes. These findings shed light on mosquito behaviour and have implications for mosquito control programs, particularly those involving the release or monitoring of the male mosquito population.

animal behavior and cognition↗

bioRxiv2022

Hidden endosymbionts: A male-killer concealed by another endosymbiont and a nuclear suppressor

Maternally transmitted endosymbiotic bacteria that cause male killing (MK) have only been described from a few insects, but this may reflect challenges in their detection rather than a rarity of MK. Here we identify MK Wolbachia in populations of Drosophila pseudotakahashii, present at a low frequency (around 4%) in natural populations and previously undetected due to a different fixed Wolbachia strain in this species expressing a different reproductive manipulation, cytoplasmic incompatibility (CI). The MK phenotype was eliminated after tetracycline treatment that removed Wolbachia. Molecular analyses indicated the MK phenotype to be expressed when a second Wolbachia strain was present alongside the CI Wolbachia. A genomic analysis highlighted Wolbachia regions diverged between the strains involving 17 genes and also identified the Wolbachia as representing an outgroup to a clade of Wolbachia infecting melanogaster-group species, including wRi-like and wMel-like strains. Doubly infected males induced CI with uninfected females but not females singly infected with CI-causing Wolbachia. The MK phenotype manifested at the larval stage and was transmitted maternally at a high fidelity but with occasional loss of the MK Wolbachia strain. A rapidly spreading dominant nuclear suppressor genetic element affecting MK was identified through backcrossing and subsequent analysis with ddRAD SNPs of the D. pseudotakahashii genome. These findings highlight the complexity of nuclear and microbial components affecting MK endosymbiont detection and dynamics in populations, and the challenges of making connections between endosymbionts and the host phenotypes affected by them.

evolutionary biology↗

bioRxiv2022

Developing Wolbachia-based disease interventions for an extreme environment

Aedes aegypti mosquitoes carrying self-spreading, virus-blocking Wolbachia bacteria are being deployed to suppress dengue transmission. However, there are challenges in applying this technology in extreme environments. We introduced two Wolbachia strains into Ae. aegypti from Saudi Arabia for a release program in the hot coastal city of Jeddah. Wolbachia reduced infection and dissemination of dengue virus (DENV2) in Saudi Arabian mosquitoes and showed complete maternal transmission and cytoplasmic incompatibility. Wolbachia reduced mosquito heat tolerance and egg viability, with the Wolbachia strains showing differential thermal stability. Wolbachia effects were similar across mosquito genetic backgrounds but we found evidence of local adaptation, with Saudi Arabian mosquitoes having lower egg viability but higher adult desiccation tolerance than Australian mosquitoes. Genetic background effects will influence Wolbachia invasion dynamics, reinforcing the need to use local genotypes for mosquito release programs, particularly in extreme environments like Jeddah. Our comprehensive characterization of Wolbachia strains provides a foundation for Wolbachia-based disease interventions in harsh climates.

evolutionary biology↗