Virulence determinants of infectious bursal disease virus

The very virulent (vv) pathotype of infectious bursal disease virus (IBDV) has spread rapidly throughout Europe, Asia, and the Middle East. Although Australia is currently unaffected, there remains the potential for incursion of an exotic isolate. The aim of this study was to identify putative virulence determinants of IBDV to facilitate the development of improved diagnostic assays for detection and characterisation of vvIBDV isolates. Sequencing of Indonesian vvIBDV Tasik94 revealed a unique substitution [ A�¨S222] in the hypervariable region (HVR) of viral protein (VP) VP2, which did not appear to impinge on virulence or antigenicity. Phylogenetic analyses indicated that Tasik94 was closely related to Asian and European vvIBDV strains. Extensive alignment of deduced protein sequences across the HVR of VP2 identified residuesI242 I256 and I294 as putative markers of the vv phcnotype. Comparison of the pathology induced by mildly-virulent Australian IBDV 002/73 and Indonesian vvIBDV Tasik94, revealed that histological lesions in the spleen, thymus and bone marrow were restricted to Tasik94-infected birds, suggesting the enhanced pathogenicity of vvIBDV might be attributed to replication in non-bursal lymphoid organs. The biological significance of the VP2 HVR in virulence was assessed using recombinant viruses generated by reverse genetics. Both genomic segments of Australian IBDV 002/73, and recombinant segment A constructs in which the HVR of 002/73 was replaced with the corresponding region of either tissue culture-adapted virus or vvIBDV (Tasik94), were cloned behind T7 RNA polymerase promoter sequences. In vitro transcription/translation of each construct resulted in expression of viral proteins. Co-transfection of synthetic RNA transcripts initiated replication of both tissue culture-adapted parental and recombinant viruses, however attempts to rescue non-adapted viruses in specific-pathogen-free (SPF) chickens were unsuccessful. Nucleotide sequence variation in the HVR of VP2 was exploited for the development of a new diagnostic assay to rapidly detect exotic IBDV isolates, including vvIBDV, using reverse transcription polymerase chain reaction (RT-PCR) amplification and Bmrl restriction enzyme digestion. The assay was capable of differentiating between endemic and exotic IBDV in 96% of 105 isolates sequenced to date.

Molecular analysis of Australian Newcastle disease virus isolates

The results presented demonstrate that outbreaks of virulent Newcastle disease in Australia arose from the emergence of virulent virus from an endemic avirulent quasispecies. Environmental selection pressures, or industrial practices, led to the emergence of this species and its dominance in several distinct ourbreaks from 1998-2002.

Characterisation and comparison of bovine RNA polymerase III promoters for RNA interference

The characterisation of novel bovine RNA polymerase III promoters for the expression of short hairpin RNAs for gene silencing resulted in the identification of a highly efficient promoter sequence. The replication of bovine viral diarrhea virus was them suppressed using short hairpin RNAs expressed form this promoter in vitro.

Design, fabrication and evaluation of universal polymerase chain reaction devices

Novel and low cost PCR devices were developed to perform rapid polymerase chain reaction based diagnostics for infectious diseases. The main advantages of using these devices are that it can perform multi sample and also multiplexing of DNA samples woth low sample volume compared to the conventional PCR devices.

Hepatitis C virus infection in South Australian prisoners : seroprevalence, seroconversion, and risk factors

Objectives : To determine entry antibody seroprevalence and seroconversion to hepatitis C virus (HCV) and associated risk factors in newly incarcerated prisoners.

Methods : Males and females entering South Australian prisons completed risk factor surveys and were offered HCV-antibody testing. Participants completed additional surveys and, if HCV-negative at last test, underwent further antibody tests at 3-monthly intervals for up to 15 months. Data were analyzed using univariate and multivariate techniques.

Results : HCV seroprevalence among 662 prison entrants was estimated at 42%. Previous injecting history was highly prevalent at entry (64%) and both community and prison injecting independently predicted entry HCV status. Tattooing was not an important risk factor. While community exposure could not be ruled out, three seroconversions were noted in 148 initially HCV-seronegative individuals occurring in a median 121 days – 4.6 per 100 person-years. Prison injecting was infrequently reported, but HCV-seropositive participants were significantly more likely to commence IDU in prison than seronegative participants (p = 0.035).

Conclusions : Entry HCV seroprevalence in South Australian prisoners is extremely high and may have contributed to a ‘ceiling effect’, minimizing the observable seroconversion rate. Greater frequency of injecting among those already infected with HCV represents a significant threat to other prisoners and prison staff.

Recent advances on the possible neuroprotective activities of Epstein-Barr virus oncogene BARF1 protein in chronic inflammatory disorders of central nervous system

Multiple sclerosis and neurodegenerative diseases in which cells of the central nervous system (CNS) are lost or damaged are rapidly increasing in frequency, and there is neither effective treatment nor cure to impede or arrest their destructive course. The Epstein-Barr virus is a human gamma-herpesvirus that infects more than 90% of the human population worldwide and persisting for the lifetime of the host. It is associated with numerous epithelial cancers, principally undifferentiated nasopharyngeal carcinoma and gastric carcinoma. Individuals with a history of symptomatic primary EBV infection, called infectious mononucleosis, carry a moderately higher risk of developing multiple sclerosis (MS). It is not known how EBV infection potentially promotes autoimmunity and central nervous system (CNS) tissue damage in MS. Recently it has been found that EBV isolates from different geographic regions have highly conserved BARF1 epitopes. BARF1 protein has the neuroprotective and mitogenic activity, thus may be useful to combat and overcome neurodegenerative disease. BARF1 protein therapy can potentially be used to enhance the neuroprotective activities by combinational treatment with anti-inflammatory antagonists and neuroprotectors in neural disorders.

Surveillance of wild birds for avian influenza virus

Recent demand for increased understanding of avian influenza virus in its natural hosts, together with the development of high-throughput diagnostics, has heralded a new era in wildlife disease surveillance. However, survey design, sampling, and interpretation in the context of host populations still present major challenges. We critically reviewed current surveillance to distill a series of considerations pertinent to avian influenza virus surveillance in wild birds, including consideration of what, when, where, and how many to sample in the context of survey objectives. Recognizing that wildlife disease surveillance is logistically and financially constrained, we discuss pragmatic alternatives for achieving probability-based sampling schemes that capture this host-pathogen system. We recommend hypothesis-driven surveillance through standardized, local surveys that are, in turn, strategically compiled over broad geographic areas. Rethinking the use of existing surveillance infrastructure can thereby greatly enhance our global understanding of avian influenza and other zoonotic diseases.

Spatial patterns, landscape genetics and post virus recovery of blacklip abalone, Haliotis rubra (Leach), in the western commercial fishing zone of Victoria

Historically, collecting nearshore habitat information has been problematic. Existing methods, such as aerial and satellite image interpretation are limited due to the attenuation of light in the water column obscuring the seabed structure. The advent of airborne bathymetric LiDAR (Light Detection and Ranging) systems (laser scanning of the seabed) now provides high-resolution seabed ‘images’ in areas that were previously difficult to survey. LiDAR imagery is available for the entire coastline of Victoria, Australia to depths of around 25 m, after being initially collected for climate change modelling by the Future Coasts Program (http://www.climatechange.vic.gov.au/adapting-to-climate-change/future-coasts). This dataset has provided the opportunity to test its applicability to inform fisheries management. Detailed geophysical information combined with spatially explicit AbTrack GPS located fisheries records and targeted genetic sampling is used in this study to provide a better understanding of the extent of available fishing grounds, direction of fishing effort and stock population structure within the Victorian western zone abalone fishery.
The species distribution modelling technique MaxEnt was used to produce a potential habitat suitability map for abalone in an attempt to capture the effective footprint of the  fishery. Also, by interrogating the spatially defined effort localities, we demonstrate an approach that may be used to identify areas where fishing effort is concentrated, and how this parameter changes temporally.
Despite barriers to adult dispersal (soft sediment barriers between reef patches), the genetic study indicates that larval movement is able to homogenize the gene pool over  large geographic distances. The western, central and eastern zone abalone stocks in Victoria were found to be a single large panmictic unit. This indicates high levels of stock connectivity and no obvious impacts of Abalone Viral Ganglioneuritis (AVG) on the genetic health of western zone stocks. We used detailed seafloor structure information interpreted from LiDAR to inform a replicated hierarchical fine scale genetic sampling design. We demonstrated that there may be extensive migration among abalone stocks across the Victorian abalone fishery.
This is contrary to previous studies that suggest recruitment is highly localised. In combination, these findings provide a valuable insight into the biology of H. rubra and immediate benefits for fisheries management. We discuss these results in the context of predicting resilience and adaptive potential of H. rubra stocks to environmental pressures and the spread of heritable diseases.
Adoption pathways are also provided to benefit future stock augmentation activities to catalyse the recovery of AVG affected reef codes. As larval dispersal is likely to be spatially and temporally variable, some AVG affected stocks are likely to recover through natural recruitment, while others will benefit from augmentation activities to ‘kick-start’ stock recovery. Evidence of neutral genetic homogeneity across Victorian reef codes suggests that the relocation of animals is unlikely to have significant genetic risks; however the potential for locally adaptive genetic differences may exist, and should be taken into consideration in future stock augmentation planning.
When combined, the spatial and genetic analyses provide valuable insights into stock productivity within the western zone fishery. Reefs appear to be expansive and support much available habitat, and the movement of larvae among reef structures is likely to be extensive in this region. Consequently, we propose that colonisation success and productivity is likely to be driven by ecological factors such as resources and/or competition, or physical factors such as wave exposure.

Phylogenetic analysis of beak and feather disease virus across a host ring-species complex

Pathogens have been hypothesized to play a major role in host diversity and speciation. Susceptibility of hybrid hosts to pathogens is thought to be a common phenomenon that could promote host population divergence and subsequently speciation. However, few studies have tested for pathogen infection across animal hybrid zones while testing for codivergence of the pathogens in the hybridizing host complex. Over 8 y, we studied natural infection by a rapidly evolving single-strand DNA virus, beak and feather diseases virus (BFDV), which infects parrots, exploiting a host-ring species complex (Platycercus elegans) in Australia. We found that host subspecies and their hybrids varied strikingly in both BFDV prevalence and load: both hybrid and phenotypically intermediate subspecies had lower prevalence and load compared with parental subspecies, while controlling for host age, sex, longitude and latitude, as well as temporal effects. We sequenced viral isolates throughout the range, which revealed patterns of genomic variation analogous to Mayr's ring-species hypothesis, to our knowledge for the first time in any host-pathogen system. Viral phylogeny, geographic location, intraspecific host density, and parrot community diversity and composition did not explain the differences in BFDV prevalence or load between subpopulations. Overall, our analyses suggest that functional host responses to infection, or force of infection, differ between subspecies and hybrids. Our findings highlight the role of host hybridization and clines in altering host-pathogen interactions, dynamics that can have important implications for models of speciation with gene flow, and offer insights into how pathogens may adapt to diverging host populations.

Swine influenza virus PA and neuraminidase gene reassortment into human H1N1iInfluenza Virus is associated with an altered pathogenic phenotype linked to increased MIP-2 expression

Swine are susceptible to infection by both avian and human influenza viruses, and this feature is thought to contribute to novel reassortant influenza viruses. In this study, the influenza virus reassortment rate in swine and human cells was determined. Coinfection of swine cells with 2009 pandemic H1N1 virus (huH1N1) and an endemic swine H1N2 (A/swine/Illinois/02860/09) virus (swH1N2) resulted in a 23% reassortment rate that was independent of α2,3- or α2,6-sialic acid distribution on the cells. The reassortants had altered pathogenic phenotypes linked to introduction of the swine virus PA and neuraminidase (NA) into huH1N1. In mice, the huH1N1 PA and NA mediated increased MIP-2 expression early postinfection, resulting in substantial pulmonary neutrophilia with enhanced lung pathology and disease. The findings support the notion that swine are a mixing vessel for influenza virus reassortants independent of sialic acid distribution. These results show the potential for continued reassortment of the 2009 pandemic H1N1 virus with endemic swine viruses and for reassortants to have increased pathogenicity linked to the swine virus NA and PA genes which are associated with increased pulmonary neutrophil trafficking that is related to MIP-2 expression. IMPORTANCE: Influenza A viruses can change rapidly via reassortment to create a novel virus, and reassortment can result in possible pandemics. Reassortments among subtypes from avian and human viruses led to the 1957 (H2N2 subtype) and 1968 (H3N2 subtype) human influenza pandemics. Recent analyses of circulating isolates have shown that multiple genes can be recombined from human, avian, and swine influenza viruses, leading to triple reassortants. Understanding the factors that can affect influenza A virus reassortment is needed for the establishment of disease intervention strategies that may reduce or preclude pandemics. The findings from this study show that swine cells provide a mixing vessel for influenza virus reassortment independent of differential sialic acid distribution. The findings also establish that circulating neuraminidase (NA) and PA genes could alter the pathogenic phenotype of the pandemic H1N1 virus, resulting in enhanced disease. The identification of such factors provides a framework for pandemic modeling and surveillance.

Weak negative associations between avian influenza virus infection and movement behaviour in a key host species, the mallard Anas platyrhynchos

Animal movements may contribute to the spread of pathogens. In the case of avian influenza virus, [migratory] birds have been suggested to play a role in the spread of some highly pathogenic strains (e.g. H5N1, H5N8), as well as their low pathogenic precursors which circulate naturally in wild birds. For a better understanding of the emergence and spread of both highly pathogenic (HPAIV) and low pathogenic avian influenza virus (LPAIV), the potential effects of LPAIVs on bird movement need to be evaluated. In a key host species, the mallard Anas platyrhynchos, we tested whether LPAIV infection status affected daily local (< 100 m) and regional (> 100 m) movements by comparing movement behaviour 1) within individuals (captured and sampled at two time points) and 2) between individuals (captured and sampled at one time point). We fitted free-living adult males with GPS loggers throughout the autumn LPAIV infection peak, and sampled them for LPAIV infection at logger deployment and at logger removal on recapture. Within individuals, we found no association between LPAIV infection and daily local and regional movements. Among individuals, daily regional movements of LPAIV infected mallards in the last days of tracking were lower than those of non-infected birds. Moreover, these regional movements of LPAIV infected birds were additionally reduced by poor weather conditions (i.e. increased wind and/or precipitation and lower temperatures). Local movements of LPAIV infected birds in the first days of tracking were higher when temperature decreased. Our study thus demonstrates that bird-assisted dispersal rate of LPAIV may be lower on a regional scale than expected on the basis of the movement behaviour of non-infected birds. Our study underlines the importance of understanding the impact of pathogen infection on host movement in order to assess its potential role in the emergence and spread of infectious diseases.

Avian influenza virus dynamics in Australian wild birds

Understanding avian influenza infection dynamics in wildlife is crucial because of the possibility of virus spill over to livestock and humans. There are still knowledge gaps how different ecological and environmental factors influence infection dynamics in birds. My study highlights the importance of investigating disease dynamics in Australia.