Testing the intrinsic mechanisms driving the dynamics of Ross River Virus across Australia.

The mechanisms driving dynamics of many epidemiologically important mosquito-borne pathogens are complex, involving combinations of vector and host factors (e.g., species composition and life-history traits), and factors associated with transmission and reporting. Understanding which intrinsic mechanisms contribute most to observed disease dynamics is important, yet often poorly understood. Ross River virus (RRV) is Australia's most important mosquito-borne disease, with variable transmission dynamics across geographic regions. We used deterministic ordinary differential equation models to test mechanisms driving RRV dynamics across major epidemic centers in Brisbane, Darwin, Mandurah, Mildura, Gippsland, Renmark, Murray Bridge, and Coorong. We considered models with up to two vector species (Aedes vigilax, Culex annulirostris, Aedes camptorhynchus, Culex globocoxitus), two reservoir hosts (macropods, possums), seasonal transmission effects, and transmission parameters. We fit models against long-term RRV surveillance data (1991-2017) and used Akaike Information Criterion to select important mechanisms. The combination of two vector species, two reservoir hosts, and seasonal transmission effects explained RRV dynamics best across sites. Estimated vector-human transmission rate (average ß = 8.04x10-4per vector per day) was similar despite different dynamics. Models estimate 43% underreporting of RRV infections. Findings enhance understanding of RRV transmission mechanisms, provide disease parameter estimates which can be used to guide future research into public health improvements and offer a basis to evaluate mitigation practices.

Arthropod-Borne Virus Surveillance as a Tool to Study the Australian Mosquito Virome.

Mosquitoes (n = 4381 in 198 pools) were collected in March and April 2018 to survey the presence of West Nile virus Kunjin strain in mosquito populations around crocodile farms in the Darwin region of the Northern Territory (NT) of Australia. While no Kunjin virus was detected in these mosquitoes, we applied our viral replicative intermediates screening system termed monoclonal antibodies to viral RNA intermediates in cells or MAVRIC to this set of samples. This resulted in the detection of 28 pools with virus replicating in C6/36 mosquito cells and the identification of three insect viruses from three distinct virus classes. We demonstrate the persistence of the insect-specific flavivirus Palm Creek virus in Coquillettidia xanthogaster mosquitoes from Darwin over almost a decade, with limited genetic drift. We also detected a novel Hubei macula-like virus 3 strain in samples from two mosquito genera, suggesting the virus, for which the sequence was originally detected in spiders and soybean thrips, might be involved in a horizontal transmission cycle between arthropods and plants. Overall, these data demonstrate the strength of the optimized MAVRIC system and contribute to our general knowledge of the mosquito virome and insect viruses.

Nucleic Acid Preservation Card Surveillance Is Effective for Monitoring Arbovirus Transmission on Crocodile Farms and Provides a One Health Benefit to Northern Australia.

The Kunjin strain of West Nile virus (WNVKUN) is a mosquito-transmitted flavivirus that can infect farmed saltwater crocodiles in Australia and cause skin lesions that devalue the hides of harvested animals. We implemented a surveillance system using honey-baited nucleic acid preservation cards to monitor WNVKUN and another endemic flavivirus pathogen, Murray Valley encephalitis virus (MVEV), on crocodile farms in northern Australia. The traps were set between February 2018 and July 2020 on three crocodile farms in Darwin (Northern Territory) and one in Cairns (North Queensland) at fortnightly intervals with reduced trapping during the winter months. WNVKUN RNA was detected on all three crocodile farms near Darwin, predominantly between March and May of each year. Two of the NT crocodile farms also yielded the detection of MVE viral RNA sporadically spread between April and November in 2018 and 2020. In contrast, no viral RNA was detected on crocodile farms in Cairns during the entire trapping period. The detection of WNVKUN and MVEV transmission by FTATM cards on farms in the Northern Territory generally correlated with the detection of their transmission to sentinel chicken flocks in nearby localities around Darwin as part of a separate public health surveillance program. While no isolates of WNVKUN or MVEV were obtained from mosquitoes collected on Darwin crocodile farms immediately following the FTATM card detections, we did isolate another flavivirus, Kokobera virus (KOKV), from Culex annulirostris mosquitoes. Our studies support the use of the FTATM card system as a sensitive and accurate method to monitor the transmission of WNVKUN and other arboviruses on crocodile farms to enable the timely implementation of mosquito control measures. Our detection of MVEV transmission and isolation of KOKV from mosquitoes also warrants further investigation of their potential role in causing diseases in crocodiles and highlights a "One Health" issue concerning arbovirus transmission to crocodile farm workers. In this context, the introduction of FTATM cards onto crocodile farms appears to provide an additional surveillance tool to detect arbovirus transmission in the Darwin region, allowing for a more timely intervention of vector control by relevant authorities.

The Removal of Subterranean Stormwater Drain Sumps as Mosquito Breeding Sites in Darwin, Australia.

The Northern Territory Top End Health Service, Medical Entomology Section and the City of Darwin council carry out a joint Mosquito Engineering Program targeting the rectification of mosquito breeding sites in the City of Darwin, Northern Territory, Australia. In 2005, an investigation into potential subterranean stormwater breeding sites in the City of Darwin commenced, specifically targeting roadside stormwater side entry pits. There were 79 side entry pits randomly investigated for mosquito breeding in the Darwin suburbs of Nightcliff and Rapid Creek, with 69.6% of the pits containing water holding sumps, and 45.6% of those water holding sumps breeding endemic mosquitoes. Culex quinquefasciatus was the most common mosquito collected, accounting for 73% of all mosquito identifications, with the potential vector mosquito Aedes notoscriptus also recovered from a small number of sumps. The sumps were also considered potential dry season maintenance breeding sites for important exotic Aedes mosquitoes such as Aedes aegypti and Aedes albopictus, which are potential vectors of dengue, chickungunya and Zika virus. Overall, 1229 side entry pits were inspected in ten Darwin suburbs from 2005 to 2008, with 180 water holding sumps identified and rectified by concrete filling.

Investigation into High Barmah Forest Virus Disease Case Numbers Reported in the Northern Territory, Australia in 2012-2013.

Between October 2012 and October 2013, unprecedented high numbers of Barmah Forest virus (BFV) disease cases were reported in the Northern Territory (NT). An investigation was launched by the NT Department of Health in cooperation with the Department of Primary Industry and Fisheries and the Department of Land Resource Management to investigate possible causes for this phenomenon. The investigation included virus isolations from mosquitoes collected in Darwin urban areas, BFV antibody testing in peri-urban small mammals and a human BFV disease case series investigation of recent cases. No BFV was isolated from the 4641 mosquitoes tested, none of the mammals tested positive for BFV antibodies, and the high BFV disease case numbers did not correlate with the relatively low mosquito vector numbers trapped in 2012-2013. It was estimated that up to 89% of the 79 human cases investigated did not have an acute arboviral illness and therefore had tested falsely positive. An Alere PanBio BFV immunoglobulin M enzyme-linked immunosorbent assay test kit is generally used to test for BFV, with the BFV disease case definition based on immunoglobulin M positives only. Other jurisdictions in Australia also reported high numbers of BFV disease cases, with the majority of the cases suspected to be false positives. Therefore, current testing methods need to be revised to reflect the true numbers of BFV disease cases occurring in Australia and to provide correct diagnoses for patients.

Anthropogenic ecological change and impacts on mosquito breeding and control strategies in salt-marshes, Northern Territory, Australia.

Darwin, in the tropical north of Australia, is subject to high numbers of mosquitoes and several mosquito-borne diseases. Many of Darwin's residential areas were built in close proximity to tidally influenced swamps, where long-term storm-water run-off from nearby residences into these swamps has led to anthropogenic induced ecological change. When natural wet-dry cycles were disrupted, bare mud-flats and mangroves were transformed into perennial fresh to brackish-water reed swamps. Reed swamps provided year-round breeding habitat for many mosquito species, such that mosquito abundance was less predictable and seasonally dependent, but constant and often occurring in plague proportions. Drainage channels were constructed throughout the wetlands to reduce pooled water during dry-season months. This study assesses the impact of drainage interventions on vegetation and mosquito ecology in three salt-marshes in the Darwin area. Findings revealed a universal decline in dry-season mosquito abundance in each wetland system. However, some mosquito species increased in abundance during wet-season months. Due to the high expense and potentially detrimental environmental impacts of ecosystem and non-target species disturbance, large-scale modifications such as these are sparingly undertaken. However, our results indicate that some large scale environmental modification can assist the process of wetland restoration, as appears to be the case for these salt marsh systems. Drainage in all three systems has been restored to closer to their original salt-marsh ecosystems, while reducing mosquito abundances, thereby potentially lowering the risk of vector-borne disease transmission and mosquito pest biting problems.