River Virus Research Articles

Fine-scale genomic tracking of Ross River virus using nanopore sequencing

BackgroundRoss River virus (RRV) is Australia’s most common and widespread mosquito-transmitted arbovirus and is of significant public health concern. With increasing anthropogenic impacts on wildlife and mosquito populations, it is important that we understand how RRV circulates in its endemic hotspots to determine where public health efforts should be directed. Current surveillance methods are effective in locating the virus but do not provide data on the circulation of the virus and its strains within the environment. This study examined the ability to identify single nucleotide polymorphisms (SNPs) within the variable E2/E3 region by generating full-length haplotypes from a range of mosquito trap-derived samples.MethodsA novel tiled primer amplification workflow for amplifying RRV was developed with analysis using Oxford Nanopore Technology’s MinION and a custom ARTIC/InterARTIC bioinformatic protocol. By creating a range of amplicons across the whole genome, fine-scale SNP analysis was enabled by specifically targeting the variable region that was amplified as a single fragment and established haplotypes that informed spatial-temporal variation of RRV in the study site in Victoria.ResultsA bioinformatic and laboratory pipeline was successfully designed and implemented on mosquito whole trap homogenates. Resulting data showed that genotyping could be conducted in real time and that whole trap consensus of the viruses (with major SNPs) could be determined in a timely manner. Minor variants were successfully detected from the variable E2/E3 region of RRV, which allowed haplotype determination within complex mosquito homogenate samples.ConclusionsThe novel bioinformatic and wet laboratory methods developed here will enable fast detection and characterisation of RRV isolates. The concepts presented in this body of work are transferable to other viruses that exist as quasispecies in samples. The ability to detect minor SNPs, and thus haplotype strains, is critically important for understanding the epidemiology of viruses their natural environment.Graphical

Predictive modelling of Ross River virus using climate data in the Darling Downs.

Ross River virus (RRV) is the most common mosquito-borne infection in Australia. RRV disease is characterised by joint pain and lethargy, placing a substantial burden on individual patients, the healthcare system and economy. This burden is compounded by a lack of effective treatment or vaccine for the disease. The complex RRV disease ecology cycle includes a number of reservoirs and vectors that inhabit a range of environments and climates across Australia. Climate is known to influence humans, animals and the environment and has previously been shown to be useful to RRV prediction models. We developed a negative binomial regression model to predict monthly RRV case numbers and outbreaks in the Darling Downs region of Queensland, Australia. Human RRV notifications and climate data for the period July 2001 - June 2014 were used for model training. Model predictions were tested using data for July 2014 - June 2019. The final model was moderately effective at predicting RRV case numbers (Pearson's r = 0.427) and RRV outbreaks (accuracy = 65%, sensitivity = 59%, specificity = 73%). Our findings show that readily available climate data can provide timely prediction of RRV outbreaks.

Spatial and temporal patterns of Ross River virus in south east Queensland, Australia: identification of hot spots at the rural-urban interface

BackgroundRoss River virus (RRV) is responsible for the most common vector-borne disease of humans reported in Australia. The virus circulates in enzootic cycles between multiple species of mosquitoes, wildlife reservoir hosts and humans. Public health concern about RRV is increasing due to rising incidence rates in Australian urban centres, along with increased circulation in Pacific Island countries. Australia experienced its largest recorded outbreak of 9544 cases in 2015, with the majority reported from south east Queensland (SEQ). This study examined potential links between disease patterns and transmission pathways of RRV.MethodsThe spatial and temporal distribution of notified RRV cases, and associated epidemiological features in SEQ, were analysed for the period 2001–2016. This included fine-scale analysis of disease patterns across the suburbs of the capital city of Brisbane, and those of 8 adjacent Local Government Areas, and host spot analyses to identify locations with significantly high incidence.ResultsThe mean annual incidence rate for the region was 41/100,000 with a consistent seasonal peak in cases between February and May. The highest RRV incidence was in adults aged from 30 to 64 years (mean incidence rate: 59/100,000), and females had higher incidence rates than males (mean incidence rates: 44/100,000 and 34/100,000, respectively). Spatial patterns of disease were heterogeneous between years, and there was a wide distribution of disease across both urban and rural areas of SEQ. Overall, the highest incidence rates were reported from predominantly rural suburbs to the north of Brisbane City, with significant hot spots located in peri-urban suburbs where residential, agricultural and conserved natural land use types intersect.ConclusionsAlthough RRV is endemic across all of SEQ, transmission is most concentrated in areas where urban and peri-urban environments intersect. The drivers of RRV transmission across rural-urban landscapes should be prioritised for further investigation, including identification of specific vectors and hosts that mediate human spillover.

The geography of Ross River virus infection in South Australia, 2000-2013.

Ross River virus (RRV) disease is Australia's most common arthropod-borne disease which has an important impact on population health and productivity. The aim of this study was to identify the spatial and temporal distribution of RRV notifications during 2000-2013 in South Australia (SA). The epidemiologic patterns of RRV notifications in SA from January 2000 to December 2013 were examined at a statistical local area (SLA) level. Spatial-temporal analyses were conducted using patient-reported place of exposure to characterise the recurrence of RRV infection stratified by age and sex. During the study period, a total of 3,687 RRV disease notifications were recorded in the state with state-wide mean annual rates of 16.8 cases per 100,000 persons and a 1:1.32 male:female ratio. The SLAs reporting cases of RRV disease exhibited spatial and temporal variation. Notified cases of RRV disease occurred more frequently in summer and autumn. A geographic expansion was observed of the area within which RRV cases occur. The comparison of age- and sex-standardised incidence rates, calculated by place of residence and patient-reported place of exposure, highlights the importance of using the latter to accurately display geospatial disease trends over time. Areas with the largest proportion of visitor cases and having the highest risk were mostly along the River Murray, which provides many vector mosquito habitats. Although public health interventions should be considered in all SLAs where RRV occurs, we suggest that priority should be given to the Riverland areas identified as highest risk.

First evidence of concurrent enzootic and endemic transmission of Ross River virus in the absence of marsupial reservoirs in Fiji

BackgroundRoss River virus (RRV) is a zoonotic alphavirus transmitted by several mosquito species. Until recently, endemic transmission was only considered possible in the presence of marsupial reservoirs. MethodsRRV seroprevalence was investigated in placental mammals (including horses, cows, goats, pigs, dogs, rats, and mice) in Fiji, where there are no marsupials. A total of 302 vertebrate serum samples were collected from 86 households from 10 communities in Western Fiji. ResultsNeutralizing antibodies against RRV were detected in 28% to 100% of sera depending on the species, and neutralization was strong even at high dilutions. ConclusionsThese results are unlikely to be due to cross-reactions. Chikungunya is the only other alphavirus known to be present in the Pacific Islands, but it rarely spills over into non-humans, even during epidemics. The study findings, together with a recent report of high RRV seroprevalence in humans, strongly suggest that RRV is circulating in Fiji in the absence of marsupial reservoirs. Considering that all non-human vertebrates present in Fiji are pan-global in distribution, RRV has the potential to further expand its geographic range. Further surveillance of RRV and access to RRV diagnostics will be critical for the early detection of emergence and outbreaks.

Viral Zoonoses

Human infection by zoonotic viruses—pathogens that normally infect animals—may result in no obvious illness, a nonspecific viral syndrome, or more severe illness typically characterized by hemorrhagic fever, encephalitis, or rash arthralgia. Transmission usually occurs by direct contact with or a bite from an infected animal or arthropod. Viral families discussed include Flaviviridae, Bunyaviridae, Arenaviridae, Filoviridae, Togaviridae, Rhabdoviridae, Paramyxoviridae, and Reoviridae, with consideration given to the epidemiology, diagnosis, treatment, and prevention of specific viruses. Hemorrhagic fevers addressed include dengue fever, dengue hemorrhagic fever, yellow fever, Crimean-Congo hemorrhagic fever, and Rift Valley fever; hantavirus infections; and the Marburg and Ebola viruses. Encephalitic fever–causing viruses discussed include La Crosse; Japanese; Murray Valley; St. Louis; tick-borne; West Nile; Powassan; eastern, western, and Venezuelan equine; rabies; Nipah; Barmah Forest; and Colorado tick fever. Rash arthralgia may be caused by the Barmah Forest, Chikungunya, Mayaro, O’nyong-nyong, Ross River, and dengue viruses. Other viral zoonoses considered include monkey B virus, ruminant and primate poxvirus, Newcastle, and foot-and-mouth diseases, as well as vesicular stomatitis virus infection. A diagram depicts the generalized arbovirus maintenance cycle. Tables list the important viral zoonoses that cause human disease, the principal hantaviruses that cause human disease, the arenaviruses that cause significant human illness, and the viral zoonoses endemic in the United States. This review contains 1 figure, 32 tables, and 80 references. Key words: dengue, diagnosis, encephalitis, epidemic, epidemiology, infection, rabies, virus, vaccine

Viral Zoonoses

Human infection by zoonotic viruses—pathogens that normally infect animals—may result in no obvious illness, a nonspecific viral syndrome, or more severe illness typically characterized by hemorrhagic fever, encephalitis, or rash arthralgia. Transmission usually occurs by direct contact with or a bite from an infected animal or arthropod. Viral families discussed include Flaviviridae, Bunyaviridae, Arenaviridae, Filoviridae, Togaviridae, Rhabdoviridae, Paramyxoviridae, and Reoviridae, with consideration given to the epidemiology, diagnosis, treatment, and prevention of specific viruses. Hemorrhagic fevers addressed include dengue fever, dengue hemorrhagic fever, yellow fever, Crimean-Congo hemorrhagic fever, and Rift Valley fever; hantavirus infections; and the Marburg and Ebola viruses. Encephalitic fever–causing viruses discussed include La Crosse; Japanese; Murray Valley; St. Louis; tick-borne; West Nile; Powassan; eastern, western, and Venezuelan equine; rabies; Nipah; Barmah Forest; and Colorado tick fever. Rash arthralgia may be caused by the Barmah Forest, Chikungunya, Mayaro, O’nyong-nyong, Ross River, and dengue viruses. Other viral zoonoses considered include monkey B virus, ruminant and primate poxvirus, Newcastle, and foot-and-mouth diseases, as well as vesicular stomatitis virus infection. A diagram depicts the generalized arbovirus maintenance cycle. Tables list the important viral zoonoses that cause human disease, the principal hantaviruses that cause human disease, the arenaviruses that cause significant human illness, and the viral zoonoses endemic in the United States. This review contains 1 figure, 32 tables, and 80 references. Key words: dengue, diagnosis, encephalitis, epidemic, epidemiology, infection, rabies, virus, vaccine

Ecological and life history traits are associated with Ross River virus infection among sylvatic mammals in Australia

BackgroundRoss River virus (RRV) is Australia’s most important arbovirus given its annual burden of disease and the relatively large number of Australians at risk for infection. This mosquito-borne arbovirus is also a zoonosis, making its epidemiology and infection ecology complex and cryptic. Our grasp of enzootic, epizootic, and zoonotic RRV transmission dynamics is imprecise largely due to a poor understanding of the role of wild mammalian hosts in the RRV system.MethodsThe current study applied a piecewise structural equation model (PSEM) toward an interspecific comparison of sylvatic Australian mammals to characterize the ecological and life history profile of species with a history of RRV infection relative to those species with no such history among all wild mammalian species surveyed for RRV infection. The effects of species traits were assessed through multiple causal pathways within the PSEM framework.ResultsSylvatic mammalian species with a history of RRV infection tended to express dietary specialization and smaller population density. These species were also characterized by a longer gestation length.ConclusionsThis study provides the first interspecific comparison of wild mammals for RRV infection and identifies some potential targets for future wildlife surveys into the infection ecology of this important arbovirus. An applied RRV macroecology may prove invaluable to the epidemiological modeling of RRV epidemics across diverse sylvatic landscapes, as well as to the development of human and animal health surveillance systems.

Predictive modelling of Ross River virus notifications in southeastern Australia.

Ross River virus (RRV) is a mosquito-borne virus endemic to Australia. The disease, marked by arthritis, myalgia and rash, has a complex epidemiology involving several mosquito species and wildlife reservoirs. Outbreak years coincide with climatic conditions conducive to mosquito population growth. We developed regression models for human RRV notifications in the Mildura Local Government Area, Victoria, Australia with the objective of increasing understanding of the relationships in this complex system, providing trigger points for intervention and developing a forecast model. Surveillance, climatic, environmental and entomological data for the period July 2000-June 2011 were used for model training then forecasts were validated for July 2011-June 2015. Rainfall and vapour pressure were the key factors for forecasting RRV notifications. Validation of models showed they predicted RRV counts with an accuracy of 81%. Two major RRV mosquito vectors (Culex annulirostris and Aedes camptorhynchus) were important in the final estimation model at proximal lags. The findings of this analysis advance understanding of the drivers of RRV in temperate climatic zones and the models will inform public health agencies of periods of increased risk.

CD8+ T cells control Ross River virus infection in musculoskeletal tissues of infected mice.

Ross River virus (RRV), chikungunya virus, and related alphaviruses cause debilitating polyarthralgia and myalgia. Mouse models of RRV and chikungunya virus have demonstrated a role for the adaptive immune response in the control of these infections. However, questions remain regarding the role for T cells in viral control, including the magnitude, location, and dynamics of CD8(+) T cell responses. To address these questions, we generated a recombinant RRV expressing the H-2(b)-restricted glycoprotein 33 (gp33) determinant derived from the glycoprotein of lymphocytic choriomeningitis virus. Using tetramers, we tracked gp33-specific CD8(+) T cells during RRV-lymphocytic choriomeningitis virus infection. We found that acute RRV infection induces activation of CD8(+) T cell responses in lymphoid and musculoskeletal tissues that peak from 10-14 d postinoculation, suggesting that CD8(+) T cells contribute to control of acute RRV infection. Mice genetically deficient for CD8(+) T cells or wild-type mice depleted of CD8(+) T cells had elevated RRV loads in skeletal muscle tissue, but not joint-associated tissues, at 14 d postinoculation, suggesting that the ability of CD8(+) T cells to control RRV infection is tissue dependent. Finally, adoptively transferred T cells were capable of reducing RRV loads in skeletal muscle tissue of Rag1(-/-) mice, indicating that T cells can contribute to the control of RRV infection in the absence of B cells and Ab. Collectively, these data demonstrate a role for T cells in the control of RRV infection and suggest that the antiviral capacity of T cells is controlled in a tissue-specific manner.

Projecting the impact of climate change on the transmission of Ross River virus: methodological challenges and research needs

Ross River virus (RRV) is the most common vector-borne disease in Australia. It is vitally important to make appropriate projections on the future spread of RRV under various climate change scenarios because such information is essential for policy-makers to identify vulnerable communities and to better manage RRV epidemics. However, there are many methodological challenges in projecting the impact of climate change on the transmission of RRV disease. This study critically examined the methodological issues and proposed possible solutions. A literature search was conducted between January and October 2012, using the electronic databases Medline, Web of Science and PubMed. Nineteen relevant papers were identified. These studies demonstrate that key challenges for projecting future climate change on RRV disease include: (1) a complex ecology (e.g. many mosquito vectors, immunity, heterogeneous in both time and space); (2) unclear interactions between social and environmental factors; and (3) uncertainty in climate change modelling and socioeconomic development scenarios. Future risk assessments of climate change will ultimately need to better understand the ecology of RRV disease and to integrate climate change scenarios with local socioeconomic and environmental factors, in order to develop effective adaptation strategies to prevent or reduce RRV transmission.

Attenuating Mutations in nsP1 Reveal Tissue-Specific Mechanisms for Control of Ross River Virus Infection

Ross River virus (RRV) is one of a group of mosquito-transmitted alphaviruses that cause debilitating, and often chronic, musculoskeletal disease in humans. Previously, we reported that replacement of the nonstructural protein 1 (nsP1) gene of the mouse-virulent RRV strain T48 with that from the mouse-avirulent strain DC5692 generated a virus that was attenuated in a mouse model of disease. Here we find that the six nsP1 nonsynonymous nucleotide differences between strains T48 and DC5692 are determinants of RRV virulence, and we identify two nonsynonymous nucleotide changes as sufficient for the attenuated phenotype. RRV T48 carrying the six nonsynonymous DC5692 nucleotide differences (RRV-T48-nsP1(6M)) was attenuated in both wild-type and Rag1(-/-) mice. Despite the attenuated phenotype, RRV T48 and RRV-T48-nsP1(6M) loads in tissues of wild-type and Rag1(-/-) mice were indistinguishable from 1 to 3 days postinoculation. RRV-T48-nsP1(6M) loads in skeletal muscle tissue, but not in other tissues, decreased dramatically by 5 days postinoculation in both wild-type and Rag1(-/-) mice, suggesting that the RRV-T48-nsP1(6M) mutant is more sensitive to innate antiviral effectors than RRV T48 in a tissue-specific manner. In vitro, we found that the attenuating mutations in nsP1 conferred enhanced sensitivity to type I interferon. In agreement with these findings, RRV T48 and RRV-T48-nsP1(6M) loads were similar in mice deficient in the type I interferon receptor. Our findings suggest that the type I IFN response controls RRV infection in a tissue-specific manner and that specific amino acid changes in nsP1 are determinants of RRV virulence by regulating the sensitivity of RRV to interferon. Arthritogenic alphaviruses, including Ross River virus (RRV), infect humans and cause debilitating pain and inflammation of the musculoskeletal system. In this study, we identified coding changes in the RRV nsP1 gene that control the virulence of RRV and its sensitivity to the antiviral type I interferon response, a major component of antiviral defense in mammals. Furthermore, our studies revealed that the effects of these attenuating mutations are tissue specific. These findings suggest that these mutations in nsP1 influence the sensitivity of RRV to type I interferon only in specific host tissues. The new knowledge gained from these studies contributes to our understanding of host responses that control alphavirus infection and viral determinants that counteract these responses.

Immuno-Chromatographic Wicking Assay for the Rapid Detection of Dengue Viral Antigens in Mosquitoes (Diptera: Culicidae)

There is a threat for dengue virus (DENV) reemergence in many regions of the world, particularly in areas where the DENV vectors, Aedes aegypti (L.) and Aedes albopictus (Skuse), are readily available. However, there are currently no accurate and reliable diagnostic methods to provide critical, real-time information for early detection of DENV within the vector populations to implement appropriate vector control and personal protective measures. In this article, we report the ability of an immuno-chromatographic assay developed by VecTOR Test Systems Inc. to detect DENV in a pool of female Aedes mosquitoes infected with any of the four viral serotypes. The DENV dipstick assay was simple to use, did not require a cold chain, and provided clear results within 30 min. It was highly specific and did not cross-react with samples spiked with West Nile, yellow fever, Japanese encephalitis, Rift Valley fever, chikungunya, Venezuelan equine encephalomyelitis, Ross River, LaCrosse, or Caraparu viruses. The DENV assay can provide real-time critical information on the presence of DENV in mosquitoes to public health personnel. Results from this assay will allow a rapid threat assessment and the focusing of vector control measures in high-risk areas.

A comparison of Aedes vigilax larval population densities and associated vegetation categories in a coastal wetland, Northern Territory, Australia

Darwin's northern suburbs border an extensive coastal reed and upper mangrove wetland recognized as an important larval habitat for Aedes vigilax (Skuse), the northern salt marsh mosquito, an established vector for Ross River and Barmah Forest viruses and an appreciable pest species. We sought to identify the most important vegetation categories associated with Ae. vigilax breeding to maximize the efficiency of mosquito control efforts. Using a generalized linear model with negative binominal distribution and log link, this study compares larval densities, determined by focused dipping, between 13 discernable vegetation categories. The incidence rate ratios (RR) generated can be used to compare the magnitude of larval densities for each vegetation category, compared with the reference category. Aedes vigilax larval densities were almost ten times greater in artificial drainage areas (RR=9.82), followed by tide-affected reticulate (Sporobolus/Xerochloa) areas (RR=8.15), then Schoenoplectus/mangroves (RR=2.29), compared with the reference vegetation category "lower mangroves." Furthermore, larval densities were highest in May, due to tidal inundation, for drainage areas and tide-affected reticulates (RR=12.2, 11.7, respectively) compared with March, the reference month. Thus, to maximize the efficiency of aerial salt marsh mosquito control operations in this wetland, larval control is best accomplished by concentrating on drains, Schoenoplectus/mangroves, and tide-affected reticulate areas, commencing early after the wet season. These results should apply to other areas of salt marsh mosquito breeding across northern Australia.

Retroviral restriction factors: new mechanisms of innate immunity

We are interested in characterizing the cellular machinery involved in the restriction of retrovirus replication - the components of the innate or ''intrinsic'' immunity. Studies of two such systems will be presented. In the first example, we have identified components involved in transcriptional silencing of proviral DNA by Embryonic Stem (ES) cells. It has long been known that ES cells potently block provirus expression via a transacting factor that binds to the primer binding site (PBS) for proline tRNA on the murine leukemia virus genome. We have purified the silencing complex and identified TRIM28 (Kap-1), a known transcriptional silencer, as an integral component of the complex. Further, we have identified the DNA sequence-specific binding component of the complex as being ZFP809, one of the dozens of zinc finger proteins encoded in the mammalian genome. We show that expression of ZFP809 is sufficient to render even differentiated cells highly resistant to MLV infection. Furthermore, we demonstrate that ZFP809 is able to potently block transcription from DNA constructs of human T-cell lymphotropic virus-1 (HTLV-1), which use the same primer tRNA. Finally, we found that similar silencing occurs at the distinct PBS for at least one other tRNA (namely, Lys1,2) utilized by such viruses as visna and spuma. In the second example, we have identified a novel antiviral gene that prevents the accumulation of retroviral mRNAs in the cytoplasm. The gene, designated ZAP, encodes a CCCH-type zinc finger antiviral protein. A region near the 3' end of the Moloney MuLV RNA was identified as necessary and sufficient to provide sensitivity to ZAP and to target mRNAs for loss. ZAP did not affect nuclear RNA levels but dramatically shortened the half-life of the viral RNAs in the cytoplasm. ZAP also potently inhibited HIV-1 RNA accumulation. ZAP was tested for its ability to inhibit viruses from other families and was found to potently inhibit the replication of multiple members of the alphavirus genus, including Sindbis, Semliki Forest, Ross River, and Venezuelan equine encephalitis viruses. As for retroviruses, the major block to Sindbis RNA accumulation could be localized to a small portion of the genome. Recombinant ZAP was found to bind specifically to the viral RNAs in vitro, and binding efficiency correlated well with restriction activity. ZAP binds to a single subunit of the RNA exosome, the major cellular machinery for RNA turnover, suggesting that it brings the exosome to the target RNAs to degrade them.

Climate variability and Ross River virus infections in Riverland, South Australia, 1992–2004

Ross River virus (RRV) infection is the most common notifiable vector-borne disease in Australia, with around 6000 cases annually. This study aimed to examine the relationship between climate variability and notified RRV infections in the Riverland region of South Australia in order to set up an early warning system for the disease in temperate-climate regions. Notified data of RRV infections were collected by the South Australian Department of Health. Climatic variables and monthly river flow were provided by the Australian Bureau of Meteorology and South Australian Department of Water, Land and Biodiversity Conservation over the period 1992-2004. Spearman correlation and time-series-adjusted Poisson regression analysis were performed. The results indicate that increases in monthly mean minimum and maximum temperatures, monthly total rainfall, monthly mean Southern Oscillation Index and monthly flow in the Murray River increase the likelihood, but an increase in monthly mean relative humidity decreases the likelihood, of disease transmission in the region, with different time-lag effects. This study demonstrates that a useful early warning system can be developed for local regions based on the statistical analysis of readily available climate data. These early warning systems can be utilized by local public health authorities to develop disease prevention and control activities.

Control of Arbovirus Vector <I>Verrallina funerea</I> (Diptera: Culicidae) in Southeast Queensland, Australia

In Australia, the brackish water mosquito Verrallina funerea (Theobald) (Diptera: Culicidae) is a serious pest and vector of Ross River and Barmah Forest viruses in coastal areas in the northeast of the continent. We report on the first extensive evaluation of the efficacy of various pesticides against this species, including laboratory dose-response assays and small plot field trials. In the laboratory, Ve. funerea was susceptible to temephos (Abate 100E) (lethal concentration, 95% endpoint [LC95] of 1.651 ppb), Bacillus thuringiensis subsp. israelensis de Barjac (Bti) (VectoBac 12AS) (LC95 of 0.033 ppm), s-methoprene (ProLink Liquid Larvicide [PLL]) (emergence inhibition, 95% endpoint [EI95] of 2.647 ppb), and technical grade pyriproxyfen (EI95 of 0.227 ppb). In contrast, Ve.funerea was highly refractory to Bacillus sphaericus Neide (VectoLex WG) (LC95 of >855.3 ppm). The survival of adults exposed as larvae to sublethal doses of s-methoprene was reduced for those doses greater than the EI50, although sex ratios were unaffected. Field trials demonstrated that Ve. funerea was susceptible to Bti (VectoBac G at 7 kg/ha) and s-methoprene (ProLink ProSand at 4 kg/ha), with these products giving >99 and >98% control, respectively. We also observed a significant increase in immature mortality in s-methoprene-treated pools, suggesting that s-methoprene field trials based solely on emergence inhibition of pupae may underestimate the efficacy of this insecticide.

Ross River virus transmission, infection, and disease: a cross-disciplinary review.

Ross River virus (RRV) is a fascinating, important arbovirus that is endemic and enzootic in Australia and Papua New Guinea and was epidemic in the South Pacific in 1979 and 1980. Infection with RRV may cause disease in humans, typically presenting as peripheral polyarthralgia or arthritis, sometimes with fever and rash. RRV disease notifications in Australia average 5,000 per year. The first well-described outbreak occurred in 1928. During World War II there were more outbreaks, and the name epidemic polyarthritis was applied. During a 1956 outbreak, epidemic polyarthritis was linked serologically to a group A arbovirus (Alphavirus). The virus was subsequently isolated from Aedes vigilax mosquitoes in 1963 and then from epidemic polyarthritis patients. We review the literature on the evolutionary biology of RRV, immune response to infection, pathogenesis, serologic diagnosis, disease manifestations, the extraordinary variety of vertebrate hosts, mosquito vectors, and transmission cycles, antibody prevalence, epidemiology of asymptomatic and symptomatic human infection, infection risks, and public health impact. RRV arthritis is due to joint infection, and treatment is currently based on empirical anti-inflammatory regimens. Further research on pathogenesis may improve understanding of the natural history of this disease and lead to new treatment strategies. The burden of morbidity is considerable, and the virus could spread to other countries. To justify and design preventive programs, we need accurate data on economic costs and better understanding of transmission and behavioral and environmental risks.

Specific ablation of antiviral gene expression in macrophages by antibody-dependent enhancement of Ross River virus infection.

Ross River virus (RRV) is an indigenous Australian arthropod-borne alphavirus responsible for epidemic polyarthritis (EPA), myalgia, and lethargy in humans. Macrophages and monocytes have been associated with human RRV disease, and previous studies have shown that RRV is capable of infecting macrophages via both a natural virus receptor and by Fc receptor-mediated antibody-dependent enhancement (ADE). Similar to other viruses, such as human immunodeficiency virus and dengue virus, ADE infection results in dramatic RRV growth increases for in vitro macrophage cultures. This study demonstrates that RRV could resist lipopolysaccharide (LPS)-induced antiviral activity in macrophage cultures when infection was via the ADE pathway. Investigation of this infection pathway found that RRV was able to suppress the transcription and translation of key antiviral genes (tumor necrosis factor and inducible nitric oxide synthase) in LPS-stimulated macrophages by disrupting the transcription into mRNA of the genes coding for the associated transcription factors IRF-1 and NF-kappaB. The transcription of non-antiviral control genes was not perturbed by RRV-ADE infection, and de novo protein synthesis also was not significantly affected in RRV-ADE infected cells. The ADE pathway of infection allowed RRV to specifically target antiviral genes in macrophages, resulting in unrestricted virus replication. As ADE has been observed for several virus families and associated with disease and adverse vaccination outcomes, these findings may have broad relevance to viral disease formation and antiviral vaccination strategies.

Synthesis of Tacaribe virus polypeptides in an in vitro coupled transcription and translation system

We have analyzed polypeptides synthesized in a coupled in vitro transcription and translation system in response to detergent-disrupted Tacaribe virus. Analysis of the major Tacaribe virus-specified product by two-dimensional polyacrylamide gel electrophoresis indicated that it had an isoelectric point similar to that of the Tacaribe nucleocapsid polypeptide N; however, the in vitro product had an approximate mol. wt. of 73 000, compared to a mol. wt. of 68 000 for the N protein. The 73 000 dalton product was found to yield proteolytic cleavage products with similar electrophoretic mobilities to those obtained from the virion P and N proteins. These results, as well as pulse-chase experiments in Tacaribe virus-infected cells, suggest that a 73 000 dalton polypeptide may be processed to yield the N polypeptide. The polypeptides synthesized in the coupled system depended on the amount and type of virus added; addition of purified Shark River (SR) virus, a member of the Patois group of bunyaviruses, resulted in synthesis of a polypeptide of mol. wt. 22 000 which corresponds to the SR nucleocapsid protein.