Generating prophylactic immunity against arboviruses in vertebrates and invertebrates.

The World Health Organization recently declared a global initiative to control arboviral diseases. These are mainly caused by pathogenic flaviviruses (such as dengue, yellow fever and Zika viruses) and alphaviruses (such as chikungunya and Venezuelan equine encephalitis viruses). Vaccines represent key interventions for these viruses, with licensed human and/or veterinary vaccines being available for several members of both genera. However, a hurdle for the licensing of new vaccines is the epidemic nature of many arboviruses, which presents logistical challenges for phase III efficacy trials. Furthermore, our ability to predict or measure the post-vaccination immune responses that are sufficient for subclinical outcomes post-infection is limited. Given that arboviruses are also subject to control by the immune system of their insect vectors, several approaches are now emerging that aim to augment antiviral immunity in mosquitoes, including Wolbachia infection, transgenic mosquitoes, insect-specific viruses and paratransgenesis. In this Review, we discuss recent advances, current challenges and future prospects in exploiting both vertebrate and invertebrate immune systems for the control of flaviviral and alphaviral diseases.

Automated differentiation of mixed populations of free-flying female mosquitoes under semi-field conditions.

Great advances in automated identification systems, or 'smart traps', that differentiate insect species have been made in recent years, yet demonstrations of field-ready devices under free-flight conditions remain rare. Here, we describe the results of mixed-species identification of female mosquitoes using an advanced optoacoustic smart trap design under free-flying conditions. Point-of-capture classification was assessed using mixed populations of congeneric (Aedes albopictus and Aedes aegypti) and non-congeneric (Ae. aegypti and Anopheles stephensi) container-inhabiting species of medical importance. Culex quinquefasciatus, also common in container habitats, was included as a third species in all assessments. At the aggregate level, mixed collections of non-congeneric species (Ae. aegypti, Cx. quinquefasciatus, and An. stephensi) could be classified at accuracies exceeding 90% (% error = 3.7-7.1%). Conversely, error rates increased when analysing individual replicates (mean % error = 48.6; 95% CI 8.1-68.6) representative of daily trap captures and at the aggregate level when Ae. albopictus was released in the presence of Ae. aegypti and Cx. quinquefasciatus (% error = 7.8-31.2%). These findings highlight the many challenges yet to be overcome but also the potential operational utility of optoacoustic surveillance in low diversity settings typical of urban environments.

Investigation on key aspects of mating biology in the mosquito Aedes koreicus.

Aedes koreicus Edwards, 1917 (Hulecoetomyia koreica) is a mosquito (Diptera: Culicidae) from Northeast Asia with a rapidly expanding presence outside its original native range. Over the years, the species has been discovered in several new countries, either spreading after first introduction or remaining localised to limited areas. Notably, recent studies have demonstrated the ability of the species to transmit zoonotic parasites and viruses both in the field and in laboratory settings. Combined with its invasive potential, the possible role of Ae. koreicus in pathogen transmission highlights the public health risks resulting from its invasion. In this study, we used a recently established population from Italy to investigate aspects of biology that influence reproductive success in Ae. koreicus: autogeny, mating behaviour, mating disruption by the sympatric invasive species Aedes albopictus Skuse, 1894, and the presence of the endosymbiont Wolbachia pipientis Hertig, 1936. Our laboratory population did not exhibit autogenic behaviour and required a bloodmeal to complete its ovarian cycle. When we exposed Ae. koreicus females to males of Ae. albopictus, we observed repeated attempts at insemination and an aggressive, disruptive mating behaviour initiated by male Ae. albopictus. Despite this, no sperm was identified in Ae. koreicus spermathecae. Wolbachia, an endosymbiotic bacterium capable of influencing mosquito reproductive behaviour, was not detected in this Ae. koreicus population and, therefore, had no effect on Ae. koreicus reproduction.

Insecticide resistance compromises the control of Aedes aegypti in Bangladesh.

BACKGROUND: With no effective drugs or widely available vaccines, dengue control in Bangladesh is dependent on targeting the primary vector Aedes aegypti with insecticides and larval source management. Despite these interventions, the dengue burden is increasing in Bangladesh, and the country experienced its worst outbreak in 2019 with 101 354 hospitalized cases. This may be partially facilitated by the presence of intense insecticide resistance in vector populations. Here, we describe the intensity and mechanisms of resistance to insecticides commonly deployed against Ae. aegypti in Dhaka, Bangladesh. RESULTS: Dhaka Ae. aegypti colonies exhibited high-intensity resistance to pyrethroids. Using CDC bottle assays, we recorded 2-24% mortality (recorded at 24 h) to permethrin and 48-94% mortality to deltamethrin, at 10× the diagnostic dose. Bioassays conducted using insecticide-synergist combinations suggested that metabolic mechanisms were contributing to pyrethroid resistance, specifically multi-function oxidases, esterases, and glutathione S-transferases. In addition, kdr alleles were detected, with a high frequency (78-98%) of homozygotes for the V1016G mutation. A large proportion (≤ 74%) of free-flying and resting mosquitoes from Dhaka colonies survived exposure to standard applications of pyrethroid aerosols in an experimental free-flight room. Although that exposure affected the immediate host-seeking behavior of Ae. aegypti, the effect was transient in surviving mosquitoes. CONCLUSION: The intense resistance characterized in this study is likely compromising the operational effectiveness of pyrethroids against Ae. aegypti in Dhaka. Switching to alternative chemical classes may offer a medium-term solution, but ultimately a more sustainable and effective approach to controlling dengue vectors is required. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Immunogenicity of a single fractional intradermal dose of Japanese encephalitis live attenuated chimeric vaccine.

BACKGROUND: Japanese encephalitis (JE) is endemic in Asia and the western Pacific. Vaccination is recommended for travellers to endemic regions, but the high cost of the vaccine is a major barrier to uptake. METHODS: A quasi-experimental, pre-post intervention clinical trial without a control group was conducted to assess the immunogenicity and safety of intradermal (ID) JE vaccine. Healthy adults (18-45 years) received one dose of 0.1 mL (20% of standard dose) ID Imojev® (JE live attenuated chimeric vaccine, Sanofi-Aventis). Adverse events following immunization (AEFIs) were recorded 10 days post-vaccination. Blood samples were collected at baseline, 4 and 8 weeks post-vaccination. Neutralizing antibodies were measured using 50% plaque reduction neutralization test (PRNT50). Seroconversion was defined as PRNT50 titre ≥10. An in vitro study was also conducted to quantify the rate of decay of vaccine potency after reconstitution. RESULTS: In total, 51 participants (72.6% females, median age 31 years), all non-reactive to JE virus at baseline were enrolled. Mild and moderate AEFIs were reported by 19.6% of participants; none required medical attention or interfered with normal daily activities. All participants seroconverted at 4 weeks (GMT 249.3; 95%CI:192.8-322.5) and remained seropositive at 8 weeks (GMT 135.5; 95%CI:104.5-175.6). Vaccine potency declined at a rate of 0.14 log plaque-forming units/0.5 mL per hour. CONCLUSIONS: In healthy adults, a single 0.1 mL ID dose of Imojev was safe and immunogenic, at least in the short term. Reconstituted vials of Imojev vaccine may not retain their potency after 6 hours. Fractional JE ID vaccination could be a cheaper yet effective alternative for short-term travellers. Further studies need to investigate the immune response in a wider age range of individuals and the long-term immunogenicity of fractional JE ID vaccines. CLINICAL TRIALS REGISTRATION: ACTRN12621000024842.

Wolbachia wAlbB inhibit dengue and Zika infection in the mosquito Aedes aegypti with an Australian background.

Biological control of mosquito vectors using the endosymbiotic bacteria Wolbachia is an emerging strategy for the management of human arboviral diseases. We recently described the development of a strain of Aedes aegypti infected with the Wolbachia strain wAlbB (referred to as the wAlbB2-F4 strain) through simple backcrossing of wild type Australian mosquitoes with a wAlbB infected Ae. aegypti strain from the USA. Field releases of male wAlbB2-F4 mosquitoes resulted in the successful suppression of wild populations of mosquitoes in the trial sites by exploiting the strain's Wolbachia-induced cytoplasmic incompatibility. We now demonstrate that the strain is resistant to infection by dengue and Zika viruses and is genetically similar to endemic Queensland populations. There was a fourfold reduction in the proportion of wAlbB2-F4 mosquitoes that became infected following a blood meal containing dengue 2 virus (16.7%) compared to wild type mosquitoes (69.2%) and a 6-7 fold reduction in the proportion of wAlbB2-F4 mosquitoes producing virus in saliva following a blood meal containing an epidemic strain of Zika virus (8.7% in comparison to 58.3% in wild type mosquitoes). Restriction-site Associated DNA (RAD) sequencing revealed that wAlbB2-F4 mosquitoes have > 98% Australian ancestry, confirming the successful introduction of the wAlbB2 infection into the Australian genomic background through backcrossing. Genotypic and phenotypic analyses showed the wAlbB2-F4 strain retains the insecticide susceptible phenotype and genotype of native Australian mosquitoes. We demonstrate that the Wolbachia wAlbB2-F4, in addition to being suitable for population suppression programs, can also be effective in population replacement programs given its inhibition of virus infection in mosquitoes. The ease at which a target mosquito population can be transfected with wAlbB2, while retaining the genotypes and phenotypes of the target population, shows the utility of this strain for controlling the Ae. aegypti mosquitoes and the pathogens they transmit.

Species-specific climate Suitable Conditions Index and dengue transmission in Guangdong, China.

BACKGROUND: Optimal climatic conditions for dengue vector mosquito species may play a significant role in dengue transmission. We previously developed a species-specific Suitable Conditions Index (SCI) for Aedes aegypti and Aedes albopictus, respectively. These SCIs rank geographic locations based on their climatic suitability for each of these two dengue vector species and theoretically define parameters for transmission probability. The aim of the study presented here was to use these SCIs together with socio-environmental factors to predict dengue outbreaks in the real world. METHODS: A negative binomial regression model was used to assess the relationship between vector species-specific SCI and autochthonous dengue cases after accounting for potential confounders in Guangdong, China. The potential interactive effect between the SCI for Ae. albopictus and the SCI for Ae. aegypti on dengue transmission was assessed. RESULTS: The SCI for Ae. aegypti was found to be positively associated with autochthonous dengue transmission (incidence rate ratio: 1.06, 95% confidence interval: 1.03, 1.09). A significant interaction effect between the SCI of Ae. albopictus and the SCI of Ae. aegypti was found, with the SCI of Ae. albopictus significantly reducing the effect of the SCI of Ae. aegypti on autochthonous dengue cases. The difference in SCIs had a positive effect on autochthonous dengue cases. CONCLUSIONS: Our results suggest that dengue fever is more transmittable in regions with warmer weather conditions (high SCI for Ae. aegypti). The SCI of Ae. aegypti would be a useful index to predict dengue transmission in Guangdong, China, even in dengue epidemic regions with Ae. albopictus present. The results also support the benefit of the SCI for evaluating dengue outbreak risk in terms of vector sympatry and interactions in the absence of entomology data in future research.

Rapid detection of kdr mutation F1534C in Aedes aegypti using recombinase polymerase amplification and lateral flow dipsticks.

Insecticide resistance monitoring is essential in assessing the efficacy of vector control measures. However, gold standard PCR-based molecular analyses for insecticide resistance detection are often hindered by time-consuming sample processing, as well as considerable infrastructure and resourcing requirements. In this study, we combined a novel one-step sample preparation reagent with a rapid isothermal molecular test that detects a knock down resistance (kdr) mutation (F1534C) that enables pyrethroid resistance in Aedes aegypti mosquitoes. We trialled the rapid F1534C pyrethroid resistance test using insecticide resistant Ae. aegypti mosquito bodies and compared results to a conventional, allele-specific quantitative PCR (AS-qPCR) coupled with melt curve genotyping in corresponding mosquito heads. From a strain of Ae. aegypti established from an insecticide resistant population in Merida, Mexico (n = 27), all the mosquito bodies (n = 27) tested positive with the rapid F1534C test regardless of whether they were homozygous or heterozygous. To assess diagnostic test specificity, we confirmed that F1534 was not detected in laboratory-reared, fully susceptible Ae. aegypti mosquito bodies (n = 28) using the rapid F1534C test or the conventional AS-qPCR melt curve analysis. All corresponding mosquito heads (n = 28) were homozygous wild-type FF1534. The rapid F1534C test thus demonstrated 100% diagnostic sensitivity (95% CI: 87.23% to 100%) and 100% diagnostic specificity (95% CI: 87.66% to 100.00%) for detection of the F1534C pyrethroid resistant single nucleotide polymorphism (SNP) in both heterozygous and homozygous Ae. aegypti. In the collection of mutant mosquitoes from Mexico, CC1534 homozygous mutants occurred at a frequency of 74.1% (n = 20) and FC heterozygous mutants at a frequency of 25.9% (n = 7). The rapid F1534C test significantly reduced the sample processing and testing time from approximately 6 h for the AS-qPCR melt curve analysis to only 25 min. These results demonstrate significant potential for our approach to resistance testing as a field-based, low-resource, rapid alternative to time-consuming and expensive laboratory-based detection.

A high-quality de novo genome assembly based on nanopore sequencing of a wild-caught coconut rhinoceros beetle (Oryctes rhinoceros).

BACKGROUND: An optimal starting point for relating genome function to organismal biology is a high-quality nuclear genome assembly, and long-read sequencing is revolutionizing the production of this genomic resource in insects. Despite this, nuclear genome assemblies have been under-represented for agricultural insect pests, particularly from the order Coleoptera. Here we present a de novo genome assembly and structural annotation for the coconut rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae), based on Oxford Nanopore Technologies (ONT) long-read data generated from a wild-caught female, as well as the assembly process that also led to the recovery of the complete circular genome assemblies of the beetle's mitochondrial genome and that of the biocontrol agent, Oryctes rhinoceros nudivirus (OrNV). As an invasive pest of palm trees, O. rhinoceros is undergoing an expansion in its range across the Pacific Islands, requiring new approaches to management that may include strategies facilitated by genome assembly and annotation. RESULTS: High-quality DNA isolated from an adult female was used to create four ONT libraries that were sequenced using four MinION flow cells, producing a total of 27.2 Gb of high-quality long-read sequences. We employed an iterative assembly process and polishing with one lane of high-accuracy Illumina reads, obtaining a final size of the assembly of 377.36 Mb that had high contiguity (fragment N50 length = 12 Mb) and accuracy, as evidenced by the exceptionally high completeness of the benchmarked set of conserved single-copy orthologous genes (BUSCO completeness = 99.1%). These quality metrics place our assembly ahead of the published Coleopteran genomes, including that of an insect model, the red flour beetle (Tribolium castaneum). The structural annotation of the nuclear genome assembly contained a highly-accurate set of 16,371 protein-coding genes, with only 2.8% missing BUSCOs, and the expected number of non-coding RNAs. The number and structure of paralogous genes in a gene family like Sigma GST is lower than in another scarab beetle (Onthophagus taurus), but higher than in the red flour beetle (Tribolium castaneum), which suggests expansion of this GST class in Scarabaeidae. The quality of our gene models was also confirmed with the correct placement of O. rhinoceros among other members of the rhinoceros beetles (subfamily Dynastinae) in a phylogeny based on the sequences of 95 protein-coding genes in 373 beetle species from all major lineages of Coleoptera. Finally, we provide a list of 30 candidate dsRNA targets whose orthologs have been experimentally validated as highly effective targets for RNAi-based control of several beetles. CONCLUSIONS: The genomic resources produced in this study form a foundation for further functional genetic research and management programs that may inform the control and surveillance of O. rhinoceros populations, and we demonstrate the efficacy of de novo genome assembly using long-read ONT data from a single field-caught insect.

The Emergence of Japanese Encephalitis in Australia and the Implications for a Vaccination Strategy.

Japanese encephalitis (JE) is the leading cause of viral encephalitis in Asia. Until 2022, only six locally transmitted human JE cases had been reported in Australia; five in northern Queensland and one in the Northern Territory. Thus, JE was mainly considered to be a disease of travellers. On 4 March 2022, JE was declared a 'Communicable Disease Incident of National Significance' when a locally acquired human case was confirmed in southern Queensland. By 11 May 2022, 41 human JE cases had been notified in four states in Australia, in areas where JE has never been detected before. From this perspective, we discuss the potential reasons for the recent emergence of the JE virus in Australia in areas where JE has never been previously reported as well as the implications of and options for mass immunisation programs if the outbreak escalates in a JE virus-immunologically naïve population.

Climate variability and Aedes vector indices in the southern Philippines: An empirical analysis.

BACKGROUND: Vector surveillance is an essential public health tool to aid in the prediction and prevention of mosquito borne diseases. This study compared spatial and temporal trends of vector surveillance indices for Aedes vectors in the southern Philippines, and assessed potential links between vector indices and climate factors. METHODS: We analysed routinely collected larval and pupal surveillance data from residential areas of 14 cities and 51 municipalities during 2013-2018 (House, Container, Breteau and Pupal Indices), and used linear regression to explore potential relationships between vector indices and climate variables (minimum temperature, maximum temperature and precipitation). RESULTS: We found substantial spatial and temporal variation in monthly Aedes vector indices between cities during the study period, and no seasonal trend apparent. The House (HI), Container (CI) and Breteau (BI) Indices remained at comparable levels across most surveys (mean HI = 15, mean CI = 16, mean BI = 24), while the Pupal Productivity Index (PPI) was relatively lower in most months (usually below 5) except for two main peak periods (mean = 49 overall). A small proportion of locations recorded high values across all entomological indices in multiple surveys. Each of the vector indices were significantly correlated with one or more climate variables when matched to data from the same month or the previous 1 or 2 months, although the effect sizes were small. Significant associations were identified between minimum temperature and HI, CI and BI in the same month (R2 = 0.038, p = 0.007; R2 = 0.029, p = 0.018; and R2 = 0.034, p = 0.011, respectively), maximum temperature and PPI with a 2-month lag (R2 = 0.031, p = 0.032), and precipitation and HI in the same month (R2 = 0.023, p = 0.04). CONCLUSIONS: Our findings indicated that larval and pupal surveillance indices were highly variable, were regularly above the threshold for triggering vector control responses, and that vector indices based on household surveys were weakly yet significantly correlated with city-level climate variables. We suggest that more detailed spatial and temporal analyses of entomological, climate, socio-environmental and Aedes-borne disease incidence data are necessary to ascertain the most effective use of entomological indices in guiding vector control responses, and reduction of human disease risk.

Persistence of antibodies, boostability, and interchangeability of Japanese encephalitis vaccines: A systematic review and dose-response meta-analysis.

BACKGROUND: The burden of Japanese encephalitis (JE) is substantial and is arguably one of the most serious viral encephalitic diseases with high case fatality and no specific treatment. JE vaccines are the only available mean to prevent the disease; however, the long-term persistence of antibodies, boostability, and interchangeability between different vaccine classes are not well understood. METHODS: To summarise the evidence, PubMed, Embase, and Cochrane CENTRAL were systematically searched from their inception to March 2021. Dose-response meta-analysis was utilised to synthesise the proportion of individuals who were seropositive over time after a primary vaccination course and a booster dose. Proportion meta-analysis was conducted to estimate the proportion of individuals who were seropositive as well as those who reported adverse events following a booster dose with a different vaccine class. RESULTS: Of 1053 publications retrieved, 27 studies with 4,558 participants were included. Of these, 11 studies assessed persistence of antibodies, 14 studies boostability, and 8 vaccine class interchangeability. The pooled seropositivity, 1-year after primary vaccination was 83.4% (95 %CI 78.2-89.5%) and remained stable for up to 5 years (82.7%; 95 %CI 76.1-89.4%). Rapid anamnestic response was observed 10 days post-booster dose, the proportion of individuals who were seropositive reached 96.9% (95 %CI 95.9-97.8%) and remained > 95% for up to 6 years. Inactivated mouse brain-derived vaccines followed by a booster dose of a different vaccine class was effective (i.e. seropositive 99%) and well tolerated. CONCLUSIONS: A booster dose after the primary vaccination is effective and further booster doses may be needed after 7 years. Inactivated mouse brain-derived vaccine followed by a booster with a newer vaccine class is effective and safe; although, there is a paucity of data related to newer classes of vaccines interchangeability.

Global Evolutionary History and Dynamics of Dengue Viruses Inferred from Whole Genome Sequences.

Dengue is an arboviral disease caused by dengue virus (DENV), leading to approximately 25,000 deaths/year and with over 40% of the world's population at risk. Increased international travel and trade, poorly regulated urban expansion, and warming global temperatures have expanded the geographic range and incidence of the virus in recent decades. This study used phylogenetic and selection pressure analyses to investigate trends in DENV evolution, using whole genome coding sequences from publicly available databases alongside newly sequenced isolates collected between 1963-1997 from Southeast Asia and the Pacific. Results revealed very similar phylogenetic relationships when using the envelope gene and the whole genome coding sequences. Although DENV evolution is predominantly driven by negative selection, a number of amino acid sites undergoing positive selection were found across the genome, with the majority located in the envelope and NS5 genes. Some genotypes appear to be diversifying faster than others within each serotype. The results from this research improve our understanding of DENV evolution, with implications for disease control efforts such as Wolbachia-based biocontrol and vaccine design.

The Use of Automated Traps to Assess the Efficacy of Insecticide Barrier Treatments Against Abundant Mosquitoes in Remote Environments.

Commercially available 'smart' trap technology has not yet been widely used to evaluate interventions against mosquitoes despite potential benefits. These benefits include the ability to capture data continuously at fine temporal scales without the human resources usually required for conventional trap deployment. Here, we used a commercially available smart trap (BG-Counter, Biogents) to assess the efficacy of an insecticide barrier treatment (BiFlex AquaMax) in reducing mosquito nuisance in a logistically challenging coastal environment in Queensland, Australia. Adoption of smart trap technology permitted us to conduct a uniquely detailed assessment of barrier treatments, ultimately allowing us to demonstrate significant reductions in mosquito collections from treated properties over all temporal scales. On average, daily mosquito collections from treated properties were reduced by 74.6% for the duration of the post-treatment period (56 d). This observation was supported by similar reductions (73.3%) in mosquito collections across all hours of the day. It was further found that underlying mosquito population dynamics were comparable across all study sites as evidenced by the high congruence in daily collection patterns among traps (Pearson r = 0.64). Despite limitations related to trap costs and replication, the results demonstrate that smart traps offer new precision tools for the assessment of barrier treatments and other mosquito control interventions.

Releasing incompatible males drives strong suppression across populations of wild and Wolbachia-carrying Aedes aegypti in Australia.

Releasing sterile or incompatible male insects is a proven method of population management in agricultural systems with the potential to revolutionize mosquito control. Through a collaborative venture with the "Debug" Verily Life Sciences team, we assessed the incompatible insect technique (IIT) with the mosquito vector Aedes aegypti in northern Australia in a replicated treatment control field trial. Backcrossing a US strain of Ae. aegypti carrying Wolbachia wAlbB from Aedes albopictus with a local strain, we generated a wAlbB2-F4 strain incompatible with both the wild-type (no Wolbachia) and wMel-Wolbachia Ae. aegypti now extant in North Queensland. The wAlbB2-F4 strain was manually mass reared with males separated from females using Verily sex-sorting technologies to obtain no detectable female contamination in the field. With community consent, we delivered a total of three million IIT males into three isolated landscapes of over 200 houses each, releasing ∼50 males per house three times a week over 20 wk. Detecting initial overflooding ratios of between 5:1 and 10:1, strong population declines well beyond 80% were detected across all treatment landscapes when compared to controls. Monitoring through the following season to observe the ongoing effect saw one treatment landscape devoid of adult Ae. aegypti early in the season. A second landscape showed reduced adults, and the third recovered fully. These encouraging results in suppressing both wild-type and wMel-Ae. aegypti confirms the utility of bidirectional incompatibility in the field setting, show the IIT to be robust, and indicate that the removal of this arbovirus vector from human-occupied landscapes may be achievable.

Simulating an invasion: unsealed water storage (rainwater tanks) and urban block design facilitate the spread of the dengue fever mosquito, Aedes aegypti, in Brisbane, Australia.

Aedes aegypti (Linnaeus) was once highly prevalent across eastern Australia, resulting in epidemics of dengue fever. Drought conditions have led to a rapid rise in semi-permanent, urban water storage containers called rainwater tanks known to be critical larval habitat for the species. The presence of these larval habitats has increased the risk of establishment of highly urbanised, invasive mosquito vectors such as Ae. aegypti. Here we use a spatially explicit network model to examine the role that unsealed rainwater tanks may play in population connectivity of an Ae. aegypti invasion in suburbs of Brisbane, a major Australian city. We characterise movement between rainwater tanks as a diffusion-like process, limited by a maximum distance of movement, average life expectancy, and a probability that Ae. aegypti will cross wide open spaces such as roads. The simulation model was run against a number of scenarios that examined population spread through the rainwater tank network based on non-compliance rates of tanks (unsealed or sealed) and road grids. We show that Ae. aegypti tank infestation and population spread was greatest in areas of high tank density and road lengths were shortest e.g. cul-de-sacs. Rainwater tank non-compliance rates of over 30% show increased connectivity when compared to less than 10%, suggesting rainwater tanks non-compliance should be maintained under this level to minimize the spread of an invading Ae. aegypti population. These results presented as risk maps of Ae. aegypti spread across Brisbane, can assist health and government authorities on where to optimally target rainwater tank surveillance and educational activities. Supplementary Information: The online version contains supplementary material available at 10.1007/s10530-021-02619-z.

The impact of sublethal permethrin exposure on susceptible and resistant genotypes of the urban disease vector Aedes aegypti.

BACKGROUND: In urban environments, some of the most common control tools used against the mosquito disease vector Aedes aegypti are pyrethroid insecticides applied as aerosols, fogs or residual sprays. Their efficacy is compromised by patchy deployment, aging residues, and the evolution and invasion of pyrethroid-resistant mosquitoes. A large proportion of mosquitoes in a given environment will therefore receive sublethal doses of insecticide. The potential impact of this sublethal exposure on the behaviour and biology of Ae. aegypti carrying commonly reported resistance alleles is poorly documented. RESULTS: In susceptible insects, sublethal exposure to permethrin resulted in reductions in egg viability (13.9%), blood avidity (16.7%) and male mating success (28.3%). It caused a 70% decrease in the lifespan of exposed susceptible females and a 66% decrease in the insecticide-resistant females from the parental strain. Exposure to the same dose of insecticide in the presence of the isolated kdr genotype resulted in a smaller impact on female longevity (a 58% decrease) but a 26% increase in eggs per female and a 37% increase in male mating success. Sublethal permethrin exposure reduced host-location success by 20-30% in all strains. CONCLUSION: The detrimental effects of exposure on susceptible insects were expected, but resistant insects demonstrated a less predictable range of responses, including negative effects on longevity and host-location but increases in fecundity and mating competitiveness. Overall, sublethal insecticide exposure is expected to increase the competitiveness of resistant phenotypes, acting as a selection pressure for the evolution of permethrin resistance. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The presence of knockdown resistance mutations reduces male mating competitiveness in the major arbovirus vector, Aedes aegypti.

BACKGROUND: The development of insecticide resistance in mosquitoes can have pleiotropic effects on key behaviours such as mating competition and host-location. Documenting these effects is crucial for understanding the dynamics and costs of insecticide resistance and may give researchers an evidence base for promoting vector control programs that aim to restore or conserve insecticide susceptibility. METHODS AND FINDINGS: We evaluated changes in behaviour in a backcrossed strain of Aedes aegypti, homozygous for two knockdown resistance (kdr) mutations (V1016G and S989P) isolated in an otherwise fully susceptible genetic background. We compared biting activity, host location behaviours, wing beat frequency (WBF) and mating competition between the backcrossed strain, and the fully susceptible and resistant parental strains from which it was derived. The presence of the homozygous kdr mutations did not have significant effects on blood avidity, the time to locate a host, or WBF in females. There was, however, a significant reduction in mean WBF in males and a significant reduction in estimated male mating success (17.3%), associated with the isolated kdr genotype. CONCLUSIONS: Our results demonstrate a cost of insecticide resistance associated with an isolated kdr genotype and manifest as a reduction in male mating success. While there was no recorded difference in WBF between the females of our strains, the significant reduction in male WBF recorded in our backcrossed strain might contribute to mate-recognition and mating disruption. These consequences of resistance evolution, especially when combined with other pleiotropic fitness costs that have been previously described, may encourage reversion to susceptibility in the absence of insecticide selection pressures. This offers justification for the implementation of insecticide resistance management strategies based on the rotation or alternation of different insecticide classes in space and time.

A regional suitable conditions index to forecast the impact of climate change on dengue vectorial capacity.

BACKGROUND: The mosquitoes Aedes aegypti and Ae. albopictus are the primary vectors of dengue virus, and their geographic distributions are predicted to expand further with economic development, and in response to climate change. We aimed to estimate the impact of future climate change on dengue transmission through the development of a Suitable Conditions Index (SCI), based on climatic variables known to support vectorial capacity. We calculated the SCI based on various climate change scenarios for six countries in the Asia-Pacific region (Australia, China, Indonesia, The Philippines, Thailand and Vietnam). METHODS: Monthly raster climate data (temperature and precipitation) were collected for the period January 2005 to December 2018 along with projected climate estimates for the years 2030, 2050 and 2070 using Representative Concentration Pathway (RCP) 4·5, 6·0 and 8·5 emissions scenarios. We defined suitable temperature ranges for dengue transmission of between 17·05-34·61 °C for Ae. aegypti and 15·84-31·51 °C for Ae. albopictus and then developed a historical and predicted SCI based on weather variability to measure the expected geographic limits of dengue vectorial capacity. Historical and projected SCI values were compared through difference maps for the six countries. FINDINGS: Comparing different emission scenarios across all countries, we found that most South East Asian countries showed either a stable pattern of high suitability, or a potential decline in suitability for both vectors from 2030 to 2070, with a declining pattern particularly evident for Ae. albopictus. Temperate areas of both China and Australia showed a less stable pattern, with both moderate increases and decreases in suitability for each vector in different regions between 2030 and 2070. INTERPRETATION: The SCI will be a useful index for forecasting potential dengue risk distributions in response to climate change, and independently of the effects of human activity. When considered alongside additional correlates of infection such as human population density and socioeconomic development indicators, the SCI could be used to develop an early warning system for dengue transmission.

The environmental and ecological determinants of elevated Ross River Virus exposure in koalas residing in urban coastal landscapes.

Koala populations in many areas of Australia have declined sharply in response to habitat loss, disease and the effects of climate change. Koalas may face further morbidity from endemic mosquito-borne viruses, but the impact of such viruses is currently unknown. Few seroprevalence studies in the wild exist and little is known of the determinants of exposure. Here, we exploited a large, spatially and temporally explicit koala survey to define the intensity of Ross River Virus (RRV) exposure in koalas residing in urban coastal environments in southeast Queensland, Australia. We demonstrate that RRV exposure in koalas is much higher (> 80%) than reported in other sero-surveys and that exposure is uniform across the urban coastal landscape. Uniformity in exposure is related to the presence of the major RRV mosquito vector, Culex annulirostris, and similarities in animal movement, tree use, and age-dependent increases in exposure risk. Elevated exposure ultimately appears to result from the confinement of remaining coastal koala habitat to the edges of permanent wetlands unsuitable for urban development and which produce large numbers of competent mosquito vectors. The results further illustrate that koalas and other RRV-susceptible vertebrates may serve as useful sentinels of human urban exposure in endemic areas.