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Introduction: Widespread transmission of Japanese encephalitis virus (JEV) genotype four (GIV) occurred across mainland Australia in 2022. This resulted in forty-five human cases, including seven deaths, and the identification of JEV infection in over 80 commercial piggeries. Materials and Methods: We collected mosquitoes which were trapped using CO 2 -baited light traps deployed near piggeries reporting disease or in regions linked to human cases in the Wide Bay region in the state of Queensland. Mosquitoes from four traps yielded JEV RNA by real-time RT-PCR. Pools containing RNA positive mosquitoes were inoculated onto mosquito cell monolayers. Discussion: A single isolate of JEV was obtained from a pool of mixed mosquito species. Near whole genome sequencing and phylogenetic analysis of the JEV isolate demonstrated its high genomic relatedness with JEV GIV pig sequences sampled from Queensland and the state of New South Wales in 2022. Conclusion: We report the first isolation of JEV GIV from mosquitoes collected in Australia. With only a few JEV GIV isolates available globally, the isolate we report will be essential for future research of JEV host interactions, evolution and disease markers, and development of effective therapies, vaccines, diagnostic assays, and mosquito control strategies.

Transmission of vector-borne pathogens can vary in complexity from single-vector, single-host systems through to multivector, multihost vertebrate systems. Understanding the dynamics of transmission is important for disease prevention efforts, but is dependent on disentangling complex interactions within coupled natural systems. Ross River virus (RRV) is a multivector multihost pathogen responsible for the greatest number of notified vector-borne pathogen infections in humans in Australia. Current evidence suggests that nonhuman vertebrates are critical for the maintenance and spillover of RRV into mosquito populations. Yet, there is a limited knowledge of which mosquito vector species and amplifying vertebrate host species are most important for transmission of RRV to humans. We conducted field surveys of nonhuman vertebrates and mosquitoes in the RRV endemic city of Brisbane, Australia, to assess the effect of vector and host community structure on human RRV notifications. Six suburbs were selected across a gradient of human disease notification rates. Differences in vertebrate and mosquito compositions were observed across all suburbs. Suburbs with higher RRV notification rates contained greater vertebrate biomass (dominated by the presence of horses) and higher mosquito abundances. This study suggests that horse-mosquito interactions should be considered in more detail and that vertebrate biomass and mosquito abundance be incorporated into future RRV modeling studies and considered in public health strategies for RRV management.

Since the establishment of West Nile virus (WNV) into the United States, concern has arisen that this virus may also pose a serious threat to Australian biosecurity. The vector competence of 19 Australian mosquito species for a North American strain of WNV was evaluated. Mosquitoes collected from Cairns, Brisbane, and Sydney were exposed to blood containing 10(4.0+/-0.3) cell culture infectious dose(50)/mosquito WNV that was isolated from a crow during the 1999 New York outbreak. Mosquitoes were tested 12-15 days later to determine their infection, dissemination, and transmission rates. A number of Culex spp. demonstrated a high vector competence for this virus, with some populations of Culex annulirostris, the primary Australian Kunjin virus vector, displaying transmission rates up to 84%. Similarly, Cx. quinquefasciatus and Cx. gelidus were highly competent, with infection and transmission rates of >80% and >50%, respectively. Common Aedes spp., including Aedes notoscriptus, Ae. vigilax, and Ae. procax, were moderately susceptible, and some Verrallina spp. and Coquillettidia spp. were relatively refractory to infection. Thus, Australia possesses a number of competent mosquito species that could facilitate local transmission of WNV, should it be introduced.