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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 Australian backyard mosquito, Aedes notoscriptus, is a highly urbanised pest species that has invaded New Zealand and the USA. Importantly, Ae. notoscriptus has been implicated as a vector of Ross River virus, a common and arthritogenic arbovirus in Australia, and is a laboratory vector of numerous other pathogenic viruses, including West Nile, yellow fever, and Zika viruses. To further explore live viruses harboured by field populations of Ae. notoscriptus and, more specifically, assess the genetic diversity of its virome, we processed 495 pools, comprising a total of 6,674 female Ae. notoscriptus collected across fifteen suburbs in Brisbane, Australia, between January 2018 and May 2019. Nine virus isolates were recovered and characterised by metagenomic sequencing and phylogenetics. The principal viral family represented was Flaviviridae. Known viruses belonging to the genera Flavivirus, Orbivirus, Mesonivirus, and Nelorpivirus were identified together with two novel virus species, including a divergent Thogoto-like orthomyxovirus and an insect-specific flavivirus. Among these, we recovered three Stratford virus (STRV) isolates and an isolate of Wongorr virus (WGRV), which for these viral species is unprecedented for the geographical area of Brisbane. Thus, the documented geographical distribution of STRV and WGRV, both known for their respective medical and veterinary importance, has now been expanded to include this major urban centre. Phylogenies of the remaining five viruses, namely, Casuarina, Ngewotan, the novel Thogoto-like virus, and two new flavivirus species, suggested they are insect-specific viruses. None of these viruses have been previously associated with Ae. notoscriptus or been reported in Brisbane. These findings exemplify the rich genetic diversity and viral abundance within the Ae. notoscriptus virome and further highlight this species as a vector of concern with the potential to transmit viruses impacting human or animal health. Considering it is a common pest and vector in residential areas and is expanding its global distribution, ongoing surveillance, and ecological study of Ae. notoscriptus, together with mapping of its virome and phenotypic characterisation of isolated viruses, is clearly warranted. Immanently, these initiatives are essential for future understanding of both the mosquito virome and the evolution of individual viral species.

Australia experienced its largest recorded outbreak of Ross River virus (RRV) during the 2014–15 reporting year, comprising >10,000 reported cases. We investigated epidemiologic, entomologic, and virologic factors that potentially contributed to the scale of the outbreak in Queensland, the state with the highest number of notifications (6,371). Spatial analysis of human cases showed that notifications were geographically widespread. In Brisbane, human case notifications and virus detections in mosquitoes occurred across inland and coastal locations. Viral sequence data demonstrated 2 RRV lineages (northeastern genotypes I and II) were circulating, and a new strain containing 3 unique amino acid changes in the envelope 2 protein was identified. Longitudinal mosquito collections demonstrated unusually high relative abundance of Culex annulirostris and Aedes procax mosquitoes, attributable to extensive freshwater larval habitats caused by early and persistent rainfall during the reporting year. Increased prevalence of these mosquitoes probably contributed to the scale of this outbreak.

The Australian backyard mosquito, Aedes notoscriptus, is a highly urbanised pest species that has invaded New Zealand and the USA. Importantly, Ae. notoscriptus has been implicated as a vector of Ross River virus, a common and arthritogenic arbovirus in Australia, and is a laboratory vector of numerous other pathogenic viruses, including West Nile, yellow fever, and Zika viruses. To further explore live viruses harboured by field populations of Ae. notoscriptus and, more specifically, assess the genetic diversity of its virome, we processed 495 pools, comprising a total of 6,674 female Ae. notoscriptus collected across fifteen suburbs in Brisbane, Australia, between January 2018 and May 2019. Nine virus isolates were recovered and characterised by metagenomic sequencing and phylogenetics. The principal viral family represented was Flaviviridae. Known viruses belonging to the genera Flavivirus, Orbivirus, Mesonivirus, and Nelorpivirus were identified together with two novel virus species, including a divergent Thogoto-like orthomyxovirus and an insect-specific flavivirus. Among these, we recovered three Stratford virus (STRV) isolates and an isolate of Wongorr virus (WGRV), which for these viral species is unprecedented for the geographical area of Brisbane. Thus, the documented geographical distribution of STRV and WGRV, both known for their respective medical and veterinary importance, has now been expanded to include this major urban centre. Phylogenies of the remaining five viruses, namely, Casuarina, Ngewotan, the novel Thogoto-like virus, and two new flavivirus species, suggested they are insect-specific viruses. None of these viruses have been previously associated with Ae. notoscriptus or been reported in Brisbane. These findings exemplify the rich genetic diversity and viral abundance within the Ae. notoscriptus virome and further highlight this species as a vector of concern with the potential to transmit viruses impacting human or animal health. Considering it is a common pest and vector in residential areas and is expanding its global distribution, ongoing surveillance, and ecological study of Ae. notoscriptus, together with mapping of its virome and phenotypic characterisation of isolated viruses, is clearly warranted. Immanently, these initiatives are essential for future understanding of both the mosquito virome and the evolution of individual viral species.

We describe the impact of COVID-19 mitigation measures on mosquito-borne diseases in Queensland, Australia, during the first half of 2020. Implementation of restrictions coincided with an atypical late season outbreak of Ross River virus (RRV) characterized by a peak in notifications in April (1173) and May (955) which were greater than 3-fold the mean observed for the previous four years. We propose that limitations on human movement likely resulted in the majority of RRV infections being acquired at or near the place of residence, and that an increase in outdoor activities, such as gardening and bushwalking in the local household vicinity, increased risk of exposure to RRV-infected mosquitoes. In contrast, the precipitous decline in international passenger flights led to a reduction in the number of imported dengue and malaria cases of over 70% and 60%, respectively, compared with the previous five years. This substantial reduction in flights also reduced a risk pathway for importation of exotic mosquitoes, but the risk posed by importation via sea cargo was not affected. Overall, the emergence of COVID-19 has had a varied impact on mosquito-borne disease epidemiology in Queensland, but the need for mosquito surveillance and control, together with encouragement of personal protective measures, remains unchanged.

BackgroundDisease surveillance and response are critical components of epidemic preparedness. The disease response, in most cases, is a set of reactive measures that follow the outcomes of the disease surveillance. Hence, efficient surveillance is a prerequisite for an effective response.Methodology/Principal FindingsWe introduce the principle of epidemiological soundness and utilize it to construct a disease spread network. The network implicitly quantifies the fertility (whether a case leads to secondary cases) and reproduction (number of secondary cases per infectious case) of the cases as well as the size and generations (of the infection chain) of the outbreaks. We empirically confirm that high morbidity relates positively with delay in disease response. Moreover, we identify what constitutes efficient surveillance by applying various thresholds of disease response delay to the network and report their impact on case fertility, reproduction, number of generations and ultimately, outbreak size. Lastly, we identify the components of the disease surveillance system that can be calibrated to achieve the identified efficiency thresholds.ConclusionWe identify practically achievable, efficient surveillance thresholds (on temporal scale) that lead to an effective response and identify how they can be satisfied. Our approach can be utilized to provide guidelines on spatially and demographically targeted resource allocation for public awareness campaigns as well as to improve diagnostic abilities and turn-around times for the doctors and laboratories involved.Author SummaryEfficient surveillance and effective response capabilities are pivotal to the prevention and control of the infectious diseases. The disease response is a set of reactive actions that follow the outcomes of the disease surveillance. Ergo, efficient surveillance is a perquisite for the deployment of an effective response. The quantification of the efficiency of a disease surveillance system largely depends on the epidemiological characteristics of the disease. In this paper, we introduce an approach that builds on these characteristics and measures the performance of a disease surveillance system through its impact on the incidence of the disease. Using this approach, we obtain quantitative (on a temporal scale) efficient surveillance thresholds, which if followed by a timely response, lead to a considerable reduction in the disease incidence. Furthermore, we show that these thresholds are practically achievable by identifying the obstacles that lead to less than efficient surveillance outcomes. Our approach can be applied to obtain guidelines on spatially, temporally and demographically targeted resource allocations for public awareness campaigns as well to improve diagnostic ability and turn-around times in treating doctors and pathology labs.

Background Disease surveillance and response are critical components of epidemic preparedness. The disease response, in most cases, is a set of reactive measures that follow the outcomes of the disease surveillance. Hence, timely surveillance is a prerequisite for an effective response. Methodology/principal findings We apply epidemiological soundness criteria in combination with the Latent Influence Point Process and time-to-event models to construct a disease spread network. The network implicitly quantifies the fertility (whether a case leads to secondary cases) and reproduction (number of secondary cases per infectious case) of the cases as well as the size and generations (of the infection chain) of the outbreaks. We test our approach by applying it to historic dengue case data from Australia. Using the data, we empirically confirm that high morbidity relates positively with delay in disease response. Moreover, we identify what constitutes timely surveillance by applying various thresholds of disease response delay to the network and report their impact on case fertility, reproduction, number of generations and ultimately, outbreak size. We observe that enforcing a response delay threshold of 5 days leads to a large average reduction across all parameters (occurrence 87%, reproduction 83%, outbreak size 80% and outbreak generations 47%), whereas extending the threshold to 10 days, in comparison, significantly limits the effectiveness of the response actions. Lastly, we identify the components of the disease surveillance system that can be calibrated to achieve the identified thresholds. Conclusion We identify practically achievable, timely surveillance thresholds (on temporal scale) that lead to an effective response and identify how they can be satisfied. Our approach can be utilized to provide guidelines on spatially and demographically targeted resource allocation for public awareness campaigns as well as to improve diagnostic abilities and turn-around times for the doctors and laboratories involved.

Aedes albopictus is an invasive mosquito, aggressive biter and potential disease vector. Its establishment in Europe and the Americas resulted in local disease transmission and impacted quality of outdoor recreational activities. Economic implications of its likely invasion into Australia had not been considered. A benefit-cost analysis of an elimination response scenario to an Ae . albopictus incursion in Brisbane was developed with a benefits transfer estimate of the local population's willingness-to-pay for an Ae . albopictus -free environment. Other costed considerations included i) necessary precautionary vector control responses to importations of mosquito-borne diseases (e.g. dengue, Zika) and ii) projected rises in complaints to local government regarding increased mosquito nuisance. Costs of implementing a successful elimination program were estimated between 1 and 4 AU$ per capita, and the annual benefit-cost ratio regarding the value of an unaffected outdoor environment was between 50 and 78. In the event of establishment, annual costs of local government and public health responses were between 0.5 and 1.3 AU$ per capita. The recurrent expense of permanent colonization will be magnified by loss of value associated with an Ae . albopictus -free environment. We conclude it is more cost-beneficial to conduct a thorough elimination program for Ae . albopictus than to allow establishment.

Effective vector and arbovirus surveillance requires timely and accurate screening techniques that can be easily upscaled. Next‐generation sequencing (NGS) is a high‐throughput technology that has the potential to modernize vector surveillance. When combined with DNA barcoding, it is termed ‘metabarcoding.’ The aim of our study was to establish a metabarcoding protocol to characterize pools of mosquitoes and screen them for virus. Pools contained 100 morphologically identified individuals, including one Ross River virus (RRV) infected mosquito, with three species present at different proportions: 1, 5, 94%. Nucleic acid extracted from both crude homogenate and supernatant was used to amplify a 269‐bp section of the mitochondrial cytochrome c oxidase subunit I (COI) locus. Additionally, a 67‐bp region of the RRV E2 gene was amplified from synthesized cDNA to screen for RRV. Amplicon sequencing was performed using an Illumina MiSeq, and bioinformatic analysis was performed using a DNA barcode database of Victorian mosquitoes. Metabarcoding successfully detected all mosquito species and RRV in every positive sample tested. The limits of species detection were also examined by screening a pool of 1000 individuals, successfully identifying the species and RRV from a single mosquito. The primers used for amplification, number of PCR cycles and total number of individuals present all have effects on the quantification of species in mixed bulk samples. Based on the results, a number of recommendations for future metabarcoding studies are presented. Overall, metabarcoding shows great promise for providing a new alternative approach to screening large insect surveillance trap catches.

Urban landscape features play an important role in the distribution and population spread of mosquito vectors. Furthermore, current insecticide and novel rear-and-release strategies for urban mosquito management rarely consider the spatial structure of the landscape when applying control practices. Here, we undertake a mark-recapture experiment to examine how urban features influence the movement and distribution of Australian container-inhabiting Aedes vectors. We pay attention to the role of semipermanent water storage containers, called rainwater tanks, and the influence of movement barriers, such as roads, on the spread and distribution of vector populations. Results suggest that Aedes aegypti (Linnaeus) (Diptera: Culicidae) were more likely to be captured around rainwater tanks, and that released males travel throughout residential blocks but do not cross roads. Conversely, female Aedes notoscriptus (Skuse) (Diptera: Culicidae) movement was uninhibited by roads and rainwater tanks did not influence female distribution or oviposition behavior. Using an isotropic Gaussian kernel framework, we show that vector movement is likely to be greater when applying a temporal effect, than when estimated by traditional methods. We conclude that a greater understanding on the role of urban features on vector movement will be important in the new age of rear-and-release mosquito control strategies, particularly those where estimations of movement are important for ensuring efficacy of application.

Arthropod-borne disease outbreaks, facilitated by the introduction of exotic mosquitoes, pose a significant public health threat. Recent chikungunya virus (CHIKV) epidemics in Europe highlight the importance of understanding the vector potential of invading mosquitoes. In this paper we explore the potential of Aedes koreicus, a mosquito new to Europe, to transmit CHIKV. Mosquitoes were challenged with CHIKV and maintained at two temperatures: 23 °C and a fluctuating temperature. Total CHIKV infection rates at 3, 10 and 14 days post-feeding were low for both temperature treatments (13.8% at 23 °C; 6.2% at fluctuating T). A low percentage (6.1%, n = 65) of mosquitoes maintained at a constant 23 °C showed dissemination of the virus to the wings and legs. Infection of mosquito saliva, with live virus, occurred in 2 mosquitoes. No dissemination was noted under the fluctuating temperature regime. Based on these results we conclude that CHIKV transmission by this species is possible. Refereed/Peer-reviewed

Medically important arboviruses such as dengue, Zika, and chikungunya viruses are primarily transmitted by the globally distributed mosquito Aedes aegypti. Increasing evidence suggests that transmission can be influenced by mosquito viromes. Herein RNA-Seq was used to characterize RNA metaviromes of wild-caught Ae. aegypti from Bangkok (Thailand) and from Cairns (Australia). The two mosquito populations showed a high degree of similarity in their viromes. BLAST searches of assembled contigs suggest up to 27 insect-specific viruses may infect Ae. aegypti, with up to 23 of these currently uncharacterized and up to 16 infecting mosquitoes from both Cairns and Bangkok. Three characterized viruses dominated, Phasi Charoen-like virus, Humaita-Tubiacanga virus and Cell fusing agent virus, and comparisons with other available RNA-Seq datasets suggested infection levels with these viruses may vary in laboratory-reared mosquitoes. As expected, mosquitoes from Bangkok showed higher mitochondrial diversity and carried alleles associated with knock-down resistance to pyrethroids. Blood meal reads primarily mapped to human genes, with a small number also showing homology with rat/mouse and dog genes. These results highlight the wide spectrum of data that can be obtained from such RNA-Seq analyses, and suggests differing viromes may need to be considered in arbovirus vector competence studies.

Background Aedes (Finlaya) koreicus (Edwards) is a mosquito that has recently entered Europe from Asia. This species is considered a potential threat to newly colonized territories, but little is known about its capacity to transmit pathogens or ability to compete with native mosquito species. The establishment of a laboratory colony is a necessary first step for further laboratory studies on the biology, ecology and vector competence of Ae. koreicus. Results A self-mating colony was established at QIMR Berghofer Medical Research Institute (Brisbane, Australia) from eggs of the F1 progeny of individuals collected as free-living larvae in northeastern Italy (Belluno province). Mosquitoes are currently maintained on both defibrinated sheep blood provided via an artificial membrane system and human blood from volunteers. Larvae are maintained in rain water and fed with Tetramin® fish food (©2015 Spectrum Brands - Pet, Home and Garden Division, Tetra-Fish). Morphometric measurements related to body size were taken and a fecundity index, based on wing length, was calculated. An in vivo technique for differentiating male and female pupae has been optimized. Our findings provide the basis for further studies on the ecology and physiology of Ae. koreicus. Conclusion We describe the establishment of an Ae. koreicus colony in the laboratory and identify critical requirements for the maintenance of this mosquito species under artificial conditions. The laboratory colony will facilitate studies investigating the vector potential of this species for human pathogens.

Background Indoor residual spraying is key to dengue control in Cairns and other parts of northern Queensland, Australia, where Aedes aegypti is prevalent, but the strategy faces challenges with regards to slow application time and, therefore, community coverage. A faster potential improvement might be the use of polyethylene netting impregnated with the volatile pyrethroid metofluthrin (SumiOne™). This formulation was assessed in rooms in three houses in Cairns, Australia. One emanator was placed in each room and cages of 10 female Aedes aegypti were exposed at distances of 1 and 3 m. Knockdown and landings on a human hand were counted before metofluthrin exposure and at 10, 30, 60, 90 and 120 min during exposure. In addition, two trials continued over 48 h of exposure to assess the long-term sublethal effects of metofluthrin on caged mosquitoes. Results Percentage landing rates fell to 0–2.5% in the first 10 min of exposure. Knockdown was most evident between 10 and 30 min (54% at 1 m and 33% at 3 m). Distance from the emanator strongly affected the results: mosquitoes at 3 m exhibited less knockdown and more landings than those at 1 m. As room volume increased, knockdown decreased and the number of landing increased. There is a cumulative mortality and landing inhibition and, for mosquitoes exposed to metofluthrin for > 48 h, mortality was 100% at 1 m and 90% at 3 m. Of those still alive, a small number continued to land and bite. After being removed from metofluthrin-treated rooms, exposed insect cages were found to reducing landing rates for up to 2 h. Conclusions Despite only moderate levels of knockdown during the initial hours of exposure, metofluthrin emanators were effective in reducing mosquito landing rates, especially within 1 m, even when exposed on an open veranda. The evaluation methods and results described in this paper will help inform the optimal conditions of deployment of metofluthrin emanators. These devices have the potential to reduce contact between humans and urban disease vectors faster than indoor residual spraying so supplement our current arsenal of dengue control tools.

A key determinant of insect persistence in marginal habitats is the ability to tolerate environmental extremes such as temperature.Aedes aegyptiis highly invasive and little is known about the physiological sensitivity of the species to fluctuating temperature regimes at the lower critical threshold. This has implications that limit establishment and persistence of the species in sub-optimal regions. Daily winter temperatures were measured in common Australian larval habitats, replicated in environmental chambers, and used to investigate the effect of fluctuating temperatures on the development and survival of tropical and subtropical strains of AustralianAe. aegypti. Development was slow for all treatments but both strains were able to complete development to the adult stage, suggesting previous models underestimate the potential for the species to persist in eastern Australia. Results suggested that thermal buffering in large volume habitats, and water that persists for greater than 32 days, will facilitate completion of the life cycle during sub-tropical winters. Furthermore, we provide a non-linear estimate of the lower critical temperature ofAe. aegyptithat suggests the current threshold may be incorrect. Our study demonstrates that the current re-introduction of water storage containers such as rainwater tanks, into major Australian population centres will increase the risk ofAe. aegyptiestablishment by permitting year-round development south of its current distribution.

Dengue is a global health concern. Growing insecticide resistance in the primary mosquito vector, Aedes aegypti, limits the effectiveness of vector control, so alternative tools are urgently needed. One approach is the use of biopesticides comprising entomopathogenic fungi, e.g., Beauveria bassiana and Metarhizium anisopliae. These fungi may decrease disease transmission by reducing mosquito vector longevity and also occur worldwide, although many isolates have not been tested for virulence against mosquitoes. Ninety-three isolates of entomopathogenic fungi representing six species (B. bassiana, M. anisopliae, Isaria fumosorosea, I. farinosa, I. flavovirescens, and Lecanicillium spp.) were screened as potential biological control agents of Aedes aegypti. A hierarchical, multi-criteria experimental design was undertaken to find suitable isolates. Initial screening was performed via in vitro assays measuring radial growth and spore persistence, eliminating isolates with poor growth or viability on ...

Entomopathogenic fungi, such as Metarhizium anisopliae and Beauveria bassiana, are being researched as alternatives to chemical adulticides to control mosquito vectors of malaria and dengue. Two cited concerns of fungal control include conidial viability and risks fungal entomopathogens pose to human health. We measured spore viability of 10 fungal isolates over 26 weeks and found a range of persistence. Three B. bassiana isolates maintained > or = 50% viability 14 weeks after application. No M. anisopliae isolate persisted longer than 1 week. To help assess risk of conidia as potential human allergens, we measured airborne conidia in enclosed environments after simulated biopesticide treatment of M. anisopliae. Conidia were detectable immediately after treatment, with concentrations of ~7000/m(3), decreasing over 48 hours to 500 conidia/m(3). At most, Metarhizium conidia comprised 2% of total visible particulate matter, falling to 0.1% by 2 days. The implications for viability of biological control of adult mosquitoes are discussed.

Disease-induced lethargy can diminish host capacity to repel or kill biting mosquitoes. We exposed house finches (Carpodacus mexicanus) to mosquitoes (Culex pipiens pipiens), repeated the experiment after inoculating finches with Mycoplasma gallisepticum, and then repeated the experiment with the same birds after curing their infections. We videotaped avian behaviors before and during mosquito exposure, identifying hosts through blood meal DNA fingerprinting. Results revealed heterogeneity in mosquito preference regardless of infection. Mosquitoes choosing between two healthy finches were more likely to feed upon the same individual bird consistently. When one bird was sick, mosquitoes exhibited no preference. Sick birds made fewer total defensive behaviors than healthy birds, but only foot stomps were associated with reduced mosquito feeding success. Our results suggest that Mycoplasma and other avian infections that alter bird defensive behavior may influence mosquito feeding patterns and transmission of arthropod-borne pathogens such as West Nile virus.

Avian defensive behavior against host-seeking arthropods influences transmission of vector-borne pathogens by affecting mosquito biting rate, either by preventing vector‐host contact or by increasing the rate of multiple host feeding. We exposed house sparrows (Passer domesticus L.) and chickens (Gallus gallus domesticus L.) to Culex pipiens pipiens L. overnight in a large observation cage and measured avian defensive behavior rates and mosquito blood-feeding success. Both bird species exhibited a range of defensive behaviors, 90% of which were foot stomps, head movements, and wing shakes. Total behavior rates increased proportionately with mosquito density in both species, increased after the first hour of mosquito exposure, and decreased as individual birds were exposed to mosquitoes multiple times. Mosquito blood-feeding success on house sparrows was high overall (82 6 5%) and independent of behavior rates. Blood-feeding success on chicks was lower (58 6 5%) and negatively correlated with defensive behavior rate after the first hour of mosquito exposure. Results revealed a higher percentage of partial blood meals on chicks (18 6 3% of all blood meals on chicks) than on house sparrows (4.9 6 3%). Birds of both species ate an average of 9.4 6 1.2% of mosquitoes, and this percentage was positively correlated with defensive behavior. High mosquito feeding success on house sparrows supports its role as a potential amplifying host of West Nile virus. Key words: chicken, Culex p. pipiens, defensive behavior, house sparrow, mosquito, West Nile virus. [Behav Ecol]

The sex pheromone component muscalure [(Z)-9-tricosene] on the cuticles of female house flies (Musca domestica L.) was quantified using gas chromatography. Laboratory and wild-strain house fly colonies were derived from poultry and dairy farms and a horse-riding stable. Muscalure levels from strain MB females were highest after 13 and 23 generations in culture, but 30–70% of females from early generations (F8–F12) had no detectable levels of muscalure. Flies

Progeny of house flies (Musca domestica L) from ten California poultry operations, three dairies and one horse-riding facility were tested for methomyl- and muscalure-treated bait resistance using up to three different assays: a topical assay, a no-choice feeding assay and a choice feeding assay. LD50 resistance factors from the topical assay, compared with a locally-derived susceptible colony, ranged between 1 and 4. LC50 resistance factors from the no-choice feeding assay ranged mostly between 2 and 5, with one value of 18. Measurable LT50 resistance ratios for female flies in the choice feeding assay ranged from 43 to 159; two populations had

Aedes aegypti (L.) (Diptera: Culicidae) is a highly invasive mosquito whose global distribution has fluctuated dramatically over the last 100 years. In Australia the distribution of Ae. aegypti once spanned the eastern seaboard, for 3,000 km north to south. However, during the 1900s this distribution markedly reduced and the mosquito disappeared from its southern range. Numerous hypotheses have been proffered for this retraction, however quantitative evidence of the mechanisms driving the disappearance are lacking. We examine historical records during the period when Ae. aegypti disappeared from Brisbane, the largest population centre in Queensland, Australia. In particular, we focus on the targeted management of Ae. aegypti by government authorities, that led to local elimination, something rarely observed in large cities. Numerous factors are likely to be responsible including the removal of larval habitat, especially domestic rainwater tanks, in combination with increased mosquito surveillance and regulatory enforcement. This account of historical events as they pertain to the elimination of Ae. aegypti from Brisbane, will inform assessments of the risks posed by recent human responses to climate change and the reintroduction of 300,000 rainwater tanks into the State over the past decade., Author summary We examined the historical role that water storage practices and the enforcement of anti-mosquito regulations played in the elimination of Aedes aegypti from Brisbane, a major urban centre in Australia. We examined changes in regulations pertaining to mosquitoes, collected government records documenting surveillance, and the response by the community to the actions of local authorities. Our findings indicate that anti-mosquito regulations, underpinned by effective implementation, were successful in gaining community support and removing the risk of mosquito presence at non-compliant properties. In particular, we argue that the removal of rainwater tanks which provided a permanent larval habitat in otherwise suboptimal environments, played a major role in the elimination of the species from Brisbane. Public Health regulations were supported by a large surveillance effort by local government health officers that were empowered to enforce legislation where necessary. Our findings are of importance to health authorities managing the ongoing expansion of Aedes populations, particularly in regions of sub-optimal climate and where water storage has become a major concern.

Background Extended drought conditions in south-east Queensland during the early 2000s have resulted in a culture of water harvesting and legislated water restrictions. Aedes notoscriptus is a container-breeding mosquito vector of Ross River and Barmah Forest viruses. Methods From 2008-2009, the larval habitats and seasonal abundance of domestic container-breeding mosquitoes were recorded from three suburbs of Brisbane. A knowledge, attitudes and practice questionnaire was administered to householders. A low-cost, desktop methodology was used to predict the proportion of shaded premises compared with front-of-property estimates. Results We highlight changes in the frequency of container categories for A. notoscriptus as a response to human behavioural changes to drought. Garden accoutrements, discarded household items and water storage containers accounted for 66.2% (525/793) of positive containers and 77.5% (73 441/94 731) of all immature mosquitoes. Of all household premises surveyed, 52.6% (550/1046) contained rainwater tanks and 29.4% (308/1046) harvested water in other containers, contrasting with a previous 1995 survey where neither category was observed. Both Premise Condition Index and shade directly correlated with positive premises. Conclusions Human response to drought has resulted in new habitats for domestic container-breeding mosquitoes. This recent trend of prolific water storage is similar to earlier years (1904-1943) in Brisbane when Aedes aegypti was present and dengue epidemics occurred.

Assessing parasite specificity to vector is crucial to understanding the emergence of vector-borne diseases and the evolution of parasite diversity. Avian malaria parasites have a cosmopolitan distribution and broad avian host range, which together predict they are vector generalists, but little is known about parasite-vector associations in the wild. We tested this prediction by asking if 5 different mosquito species, known to feed on birds and abundant in the northeastern United States, were naturally infected in the field with identical avian Plasmodium spp. lineages. Mosquitoes were not pooled but rather analyzed individually, and, possibly as a result, lineage diversity was higher than reported in previous avian malaria vector studies. Plasmodium spp. lineages were rare in Aedes canadensis and absent in Aedes aurifer and Culiseta melanura. We sequenced a standard Plasmodium cytochrome b marker from Culex pipiens pipiens, Culex restuans, and Ae. canadensis. Most Plasmodium clades were shared by Cx. pipiens and Cx. restuans. In addition, 4 individual lineages were shared by both mosquito species, including the most common lineage. One Plasmodium clade, however, was only found in Cx. restuans. We therefore found limited support for our prediction that avian Plasmodium spp. vector breadth accompanies host breadth. The association of both Culex species with most Plasmodium clades, and the presence of a single parasite lineage in 3 mosquito species representing 2 genera, suggests that avian Plasmodium species are not tightly coevolved with vector species.

Disease-induced lethargy can diminish host capacity to repel or kill biting mosquitoes. We exposed house finches (Carpodacus mexicanus) to mosquitoes (Culex pipiens pipiens), repeated the experiment after inoculating finches with Mycoplasma gallisepticum, and then repeated the experiment with the same birds after curing their infections. We videotaped avian behaviors before and during mosquito exposure, identifying hosts through blood meal DNA fingerprinting. Results revealed heterogeneity in mosquito preference regardless of infection. Mosquitoes choosing between two healthy finches were more likely to feed upon the same individual bird consistently. When one bird was sick, mosquitoes exhibited no preference. Sick birds made fewer total defensive behaviors than healthy birds, but only foot stomps were associated with reduced mosquito feeding success. Our results suggest that Mycoplasma and other avian infections that alter bird defensive behavior may influence mosquito feeding patterns and transmission of arthropod-borne pathogens such as West Nile virus.

Background Introduced Wolbachia bacteria can influence the susceptibility of Aedes aegypti mosquitoes to arboviral infections as well as having detrimental effects on host fitness. Previous field trials demonstrated that the wMel strain of Wolbachia effectively and durably invades Ae. aegypti populations. Here we report on trials of a second strain, wMelPop-PGYP Wolbachia, in field sites in northern Australia (Machans Beach and Babinda) and central Vietnam (Tri Nguyen, Hon Mieu Island), each with contrasting natural Ae. aegypti densities. Methods Mosquitoes were released at the adult or pupal stages for different lengths of time at the sites depending on changes in Wolbachia frequency as assessed through PCR assays of material collected through Biogents-Sentinel (BG-S) traps and ovitraps. Adult numbers were also monitored through BG-S traps. Changes in Wolbachia frequency were compared across hamlets or house blocks. Results Releases of adult wMelPop-Ae. aegypti resulted in the transient invasion of wMelPop in all three field sites. Invasion at the Australian sites was heterogeneous, reflecting a slower rate of invasion in locations where background mosquito numbers were high. In contrast, invasion across Tri Nguyen was relatively uniform. After cessation of releases, the frequency of wMelPop declined in all sites, most rapidly in Babinda and Tri Nguyen. Within Machans Beach the rate of decrease varied among areas, and wMelPop was detected for several months in an area with a relatively low mosquito density. Conclusions These findings highlight challenges associated with releasing Wolbachia-Ae. aegypti combinations with low fitness, albeit strong virus interference properties, as a means of sustainable control of dengue virus transmission. Electronic supplementary material The online version of this article (doi:10.1186/s13071-015-1174-x) contains supplementary material, which is available to authorized users.