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BACKGROUND: Murray Valley encephalitis virus (MVEV) is a flavivirus that occurs in Australia and New Guinea. While clinical cases are uncommon, MVEV can cause severe encephalitis with high mortality. Sentinel chicken surveillance is used at many sites around Australia to provide an early warning system for risk of human infection in areas that have low population density and geographical remoteness. MVEV in Western Australia occurs in areas of low population density and geographical remoteness, resulting in logistical challenges with surveillance systems and few human cases. While epidemiological data has suggested an association between rainfall and MVEV activity in outbreak years, it has not been quantified, and the association between rainfall and sporadic cases is less clear. In this study we analysed 22 years of sentinel chicken and human case data from Western Australia in order to evaluate the effectiveness of sentinel chicken surveillance for MVEV and assess the association between rainfall and MVEV activity. METHODS: Sentinel chicken seroconversion, human case and rainfall data from the Kimberley and Pilbara regions of Western Australia from 1990 to 2011 were analysed using negative binomial regression. Sentinel chicken seroconversion and human cases were used as dependent variables in the model. The model was then tested against sentinel chicken and rainfall data from 2012 and 2013. RESULTS: Sentinel chicken seroconversion preceded all human cases except two in March 1993. Rainfall in the prior three months was significantly associated with both sentinel chicken seroconversion and human cases across the regions of interest. Sentinel chicken seroconversion was also predictive of human cases in the models. The model predicted sentinel chicken seroconversion in the Kimberley but not in the Pilbara, where seroconversions early in 2012 were not predicted. The latter may be due to localised MVEV activity in isolated foci at dams, which do not reflect broader virus activity in the region. CONCLUSIONS: We showed that rainfall and sentinel chickens provide a useful early warning of MVEV risk to humans across endemic and epidemic areas, and that a combination of the two indicators improves the ability to assess MVEV risk and inform risk management measures.

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Edge Hill virus (EHV) is a mosquito-borne flavivirus isolated throughout Australia during mosquito surveillance programs. While not posing an immediate threat to the human population, EHV is a taxonomically interesting flavivirus since it remains the only member of the yellow fever virus (YFV) sub-group to be detected within Australia. Here we present both an antigenic and genetic investigation of collected isolates, and confirm taxonomic classification of the virus within the YFV-group. Isolates were not clustered based on geographical origin or time of isolation, suggesting that minimal genetic evolution of EHV has occurred over geographic distance or time within the EHV cluster. However, two isolates showed significant differences in antigenic reactivity patterns, and had a much larger divergence from the EHV prototype (19% nucleotide and 6% amino acid divergence), indicating a distinct subtype or variant within the EHV subgroup.

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Murray Valley encephalitis virus (MVEV) is a medically important mosquito-borne flavivirus found in Australia and Papua New Guinea (PNG). Partial envelope gene nucleotide sequences of 28 isolates of MVEV from Western Australia (WA) between 1972 and 2003 were aligned and compared phylogenetically with the prototype MVE-1-51 from Victoria in 1951 and isolates from northern Queensland and PNG. Monoclonal antibody-binding patterns were also investigated. Results showed that the majority of isolates of MVEV from widely disparate locations in WA were genetically and phenotypically homogeneous. Furthermore, isolates of MVEV from WA and northern Queensland were almost identical, confirming results from earlier studies. Recent isolates of MVEV from Western Province in PNG were more similar to Australian isolates of MVEV than to isolates from PNG in 1956 and 1966, providing further evidence for the movement of flaviviruses between PNG and Australia. Additional representatives of a unique variant of MVEV (OR156) from Kununurra in the northeast Kimberley region of WA were also detected. This suggests that the OR156 lineage is still intermittently active but may be restricted to a small geographic area in northern WA, possibly due to altered biological characteristics.

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We describe the development of an epitope-blocking enzyme-linked immunosorbent assay (ELISA) for the sensitive and rapid detection of antibodies to Ross River virus (RRV) in human sera and known vertebrate host species. This ELISA provides an alternative method for the serodiagnosis of RRV infections.

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Alfuy virus (ALFV) is classified as a subtype of the flavivirus Murray Valley encephalitis virus (MVEV); however, despite preliminary reports of antigenic and ecological similarities with MVEV, ALFV has not been associated with human disease. Here, it was shown that ALFV is at least 10(4)-fold less neuroinvasive than MVEV after peripheral inoculation of 3-week-old Swiss outbred mice, but ALFV demonstrates similar neurovirulence. In addition, it was shown that ALFV is partially attenuated in mice that are deficient in alpha/beta interferon responses, in contrast to MVEV which is uniformly lethal in these mice. To assess the antigenic relationship between these viruses, a panel of monoclonal antibodies was tested for the ability to bind to ALFV and MVEV in ELISA. Although the majority of monoclonal antibodies recognized both viruses, confirming their antigenic similarity, several discriminating antibodies were identified. Finally, the entire genome of the prototype strain of ALFV (MRM3929) was sequenced and phylogenetically analysed. Nucleotide (73 %) and amino acid sequence (83 %) identity between ALFV and MVEV confirmed previous reports of their close relationship. Several nucleotide and amino acid deletions and/or substitutions with putative functional significance were identified in ALFV, including the abolition of a conserved glycosylation site in the envelope protein and the deletion of the terminal dinucleotide 5'-CU(OH)-3' found in all other members of the genus. These findings confirm previous reports that ALFV is closely related to MVEV, but also highlights significant antigenic, genetic and phenotypic divergence from MVEV. Accordingly, the data suggest that ALFV is a distinct species within the serogroup Japanese encephalitis virus.

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Aedes (Aedimorphus) vexans vexans has become widely distributed in the Australasian and Oceanic Islands zoogeographic regions, and in this paper, we describe the 1st confirmed report of Ae. vexans vexans in Australia. A total of 45 adult individuals were collected around the town of Kununurra in the northeast Kimberley region of Western Australia during the late wet seasons between 1996 and 2003. The majority (84%) was collected at trap sites in or near the Ord Stage I Irrigation Area. Aedes vexans vexans was a minor component of the mosquito population, comprising <0.1% of the total number of adults collected during the study period. The absence of Ae. vexans vexans in earlier collections suggests that it has been recently introduced into northeast Kimberley, most likely by wind currents from the Indonesian archipelago or on occasional light aircraft arriving in Kununurra from nearby islands. The collection of adult Ae. vexans vexans every year since 2001 could indicate that this mosquito has become established in the Kununurra environs, possibly facilitated by the presence of year-round breeding sites created by the irrigation area. Encephalitogenic flaviviruses have been isolated from this species in North America, Europe, and Taiwan, and the vector competence of Ae. vexans vexans for arboviruses prevalent in northern Western Australia, particularly the flaviviruses Murray Valley encephalitis and Kunjin, should be investigated.

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Detection of flavivirus seroconversions in sentinel chicken flocks located in four Australian states are used to provide an early warning of increased levels of Murray Valley encephalitis virus (MVEV) and Kunjin virus (KUNV) activity in the region. During the 2003-2004 season low levels of flavivirus activity were detected in northern Australia with both MVEV and KUNV virus activity detected in the Kimberley and Pilbara regions of Western Australia and in the Northern Territory. A single case of Murray Valley encephalitis was reported from Central Australia. MVEV activity was also detected at Minindee in western New South Wales for the first time since 2000-2001. No activity was detected in Victoria.

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This report describes the epidemiology of mosquito-borne disease in Australia for the mosquito-borne disease season 1 July 2004 to 30 June 2005. Ross River virus (RRV) infections (45%), Barmah Forest virus (BFV) infections (30%) and malaria (19%) were the most common mosquito-borne diseases reported in 2004-05. The Northern Territory had the highest rate of RRV notifications and the peak notification rate (in February 2005) was 54 per cent less than the previous season. The Northern Territory also reported the highest BFV notification rate this season, peaking in April 2005, which was the second highest reported BFV notification rate since 1998. National RRV and BFV notification rates were highest in the 45-49 year age group. There were 799 notifications of malaria in 2004-05 of which none were reported as locally acquired. This was the third highest reporting season for malaria notifications since 1998. In contrast to previous years in which Plasmodium vivax was the predominant species, Plasmodium falciparum was reported as the infecting species in 57 per cent of the malaria notifications and Plasmodium vivax for 34 per cent of cases. Children in the 5-9 year age group had the highest number of cases compared to previous years in which the peak number of cases tended to be in young adult age groups. There were four cases of Kunjin virus (KUNV) and two cases of Murray Valley encephalitis virus (MVEV) reported in 2004-05. Sentinel chicken surveillance data for flaviviruses and sentinel pig surveillance data for Japanese encephalitis virus are reported. There were 188 notifications of dengue virus infection (DENV) in 2004-05, of which 46 per cent (n=86) were reported as having been acquired overseas. Dengue serotype 4 was the most frequently reported type, accounting for 32 per cent of cases (n=60).

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In the winter of 2003 Western Australia experienced its largest epidemic of influenza for at least five years, with activity peaking in August and September. The season was short resulting in very high numbers of cases during the peak weeks. Activity in country areas followed the peak of Metropolitan activity. Influenza A virus was detected in 28.3 per cent of the sentinel samples, and influenza B in less than one per cent. Both routine and sentinel detections and the overall estimates of influenza-like illnesses (ILI) seen by general practitioners at sentinel practices peaked in August and September 2003. The combination of influenza detections and an increase in ILI seemed to be the most accurate predictor of the beginning of winter influenza activity. There was a shift in age distribution for influenza A compared with 2003. Both the sentinel surveillance and routine samples demonstrated an increase of influenza in children and young adults. The majority of influenza A isolates were identified as A/Fujian/411/2002-like, a variant of the A/Moscow strain included in the vaccine. Despite this mismatch there did not seem to have been any noticeable increase in the risk of influenza infection in the vaccinated populations from the sentinel practices, nor was there a relative increase in disease among the highly vaccinated elderly population. A number of other respiratory viruses were identified as causes of influenza-like illness in the sentinel samples. Rhinoviruses and human metapneumovirus were the most common, the latter occurring mainly in adults.

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We recently characterized three novel alphaviruses isolated from mosquitoes captured in New South Wales, Australia. Initial cross-neutralization studies revealed antigenic similarity to the Sindbis virus (SINV)-like Whataroa virus (WHAV), heretofore found only in New Zealand. Nucleotide sequence analysis showed that the WHAV-like viruses shared >99% nucleotide sequence similarity with each other, and 96-97% similarity with prototype WHAV. Enzyme-linked immunosorbent assay reactions of a panel of monoclonal antibodies to SINV showed that the novel WHAV-like viruses displayed identical binding patterns and were antigenically distinct from all SINV isolates examined. Although these viruses displayed a similar binding pattern to prototype WHAV, three monoclonal antibodies discriminated them from the New Zealand virus. Our results suggest that these novel alphaviruses are antigenic variants of WHAV and represent the first reported isolations of this virus from outside New Zealand. The monoclonal antibodies used in this study will be useful for typing new SINV and SINV-like isolates.

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We undertook annual surveys of flavivirus activity in the community of Billiluna in the southeast Kimberley region of Western Australia between 1989 and 2001 [corrected]. Culex annulirostris was the dominant mosquito species, particularly in years of above average rains and flooding. Murray Valley encephalitis (MVE) virus was isolated in 8 of the 13 years of the study from seven mosquito species, but more than 90% of the isolates were from Cx. annulirostris. The results suggest that MVE virus is epizootic in the region, w ith activity only apparent in years with average or above average rainfall and increased numbers of Cx. annulirostris. High levels of MVE virus activity and associated human cases were detected only once (in 1993) during the survey period. Activity of MVE virus could only be partially correlated with wet season rainfall and flooding, suggesting that a number of other factors must also be considered to accurately predict MVE virus activity at such communities.

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Detection of flavivirus seroconversions in sentinel chicken flocks located throughout Australia is used to provide an early warning of increased levels of Murray Valley encephalitis (MVE) and Kunjin (KUN) virus activity in the region. During the 2002-2003 season low levels of flavivirus activity were detected in northern Australia compared to previous years. MVE and KUN virus activity was detected in the Kimberley and Pilbara regions of Western Australia and the Northern Territory but not in north Queensland, New South Wales or Victoria. This is similar to the previous season. There were no reported cases of disease caused by either virus.

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Our previous studies have shown that two distinct genotypes of Sindbis (SIN) virus occur in Australia. One of these, the Oriental/Australian type, circulates throughout most of the Australian continent, whereas the recently identified south-west (SW) genetic type appears to be restricted to a distinct geographic region located in the temperate south-west of Australia. We have now determined the complete nucleotide and translated amino acid sequences of a SW isolate of SIN virus (SW6562) and performed comparative analyses with other SIN viruses at the genomic level. The genome of SW6562 is 11,569 nucleotides in length, excluding the cap nucleotide and poly (A) tail. Overall this virus differs from the prototype SIN virus (strain AR339) by 23% in nucleotide sequence and 12.5% in amino acid sequence. Partial sequences of four regions of the genome of four SW isolates were determined and compared with the corresponding sequences from a number of SIN isolates from different regions of the World. These regions are the non-structural protein (nsP3), the E2 gene, the capsid gene, and the repeated sequence elements (RSE) of the 3'UTR. These comparisons revealed that the SW SIN viruses were more closely related to South African and European strains than to other Australian isolates of SIN virus. Thus the SW genotype of SIN virus may have been introduced into this region of Australia by viremic humans or migratory birds and subsequently evolved independently in the region. The sequence data also revealed that the SW genotype contains a unique deletion in the RSE of the 3'UTR region of the genome. Previous studies have shown that deletions in this region of the SIN genome can have significant effects on virus replication in mosquito and avian cells, which may explain the restricted distribution of this genotype of SIN virus.

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Detection of flavivirus seroconversions in sentinel chicken flocks located throughout Australia is used to provide an early warning of increased levels of Murray Valley encephalitis (MVE) and Kunjin virus activity in the region. During the 2001/2002 season low levels of flavivirus activity were detected in northern Australia compared to previous years. MVE and Kunjin virus activity was detected in the Kimberley and Pilbara regions of Western Australia and the Northern Territory but not in north Queensland, New South Wales or Victoria. This is in contrast to the previous season when MVE activity was detected both in northern Australia and, for the first time in over 20 years, in New South Wales. Two cases of Murray Valley encephalitis were reported from the north of Western Australia during the 2001/2002 wet season.

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Murray Valley encephalitis (MVE) virus is a mosquito-borne flavivirus causing severe encephalitis with a resultant high morbidity and mortality. In the period 1989-1993, we undertook a cross-sectional and longitudinal study by annually screening members of a small remote Aboriginal community in northwestern Australia for MVE virus antibodies. Of the estimated 250-300 people in the community, 249 were tested, and 52.6% had positive serology to MVE. The proportion testing positive increased with increasing age group, and males were slightly more likely to be positive than females. During the study period, a high proportion of the population seroconverted to MVE; the clinical/subclinical ratio seems to be lower than previously reported. Although MVE is mostly asymptomatic, the devastating consequences of clinical illness indicate that advice should be provided regarding the avoidance of mosquito bites. Our longitudinal study showed that the risk of seroconversion was similar for each age group, not just the young.

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Western Australia experienced its worst-ever outbreak of the mosquito-borne Murray Valley encephalitis (MVE) virus during the 2000 wet season. Highest-on-record rainfall throughout much of the state during the 2000 wet season gave rise to extensive mosquito breeding and increased MVE virus transmission, resulting in nine cases of encephalitis. Activity of MVE virus in Western Australia is monitored by detecting MVE virus-specific antibodies in serum from sentinel chickens, located at towns and communities throughout the north of the state. However, during 2000, all 28 flocks of chickens seroconverted to MVE virus, including a flock located >600 km further south than MVE virus activity had ever previously been recorded. Furthermore, the majority of the nine cases of encephalitis occurred outside the enzootic Kimberley region. We therefore undertook a major serosurvey of domestic chicken flocks both south and east of the previously defined regions of virus activity. The results suggest that MVE virus activity extended as far south as the Midwest and northern Goldfields during 2000. A new southern limit of activity of MVE virus is therefore proposed. The results have implications for managing outbreaks of MVE virus in Western Australia and have enabled us to locate additional sentinel flocks as part of the MVE surveillance program for future years.

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