Routes of Hendra Virus Excretion in Naturally-Infected Flying-Foxes: Implications for Viral Transmission and Spillover Risk

Pteropid bats or flying-foxes (Chiroptera: Pteropodidae) are the natural host of Hendra virus (HeV) which sporadically causes fatal disease in horses and humans in eastern Australia. While there is strong evidence that urine is an important infectious medium that likely drives bat to bat transmission and bat to horse transmission, there is uncertainty about the relative importance of alternative routes of excretion such as nasal and oral secretions, and faeces. Identifying the potential routes of HeV excretion in flying-foxes is important to effectively mitigate equine exposure risk at the bat-horse interface, and in determining transmission rates in host-pathogen models. The aim of this study was to identify the major routes of HeV excretion in naturally infected flying-foxes, and secondarily, to identify between-species variation in excretion prevalence. A total of 2840 flying-foxes from three of the four Australian mainland species (Pteropus alecto, P. poliocephalus and P. scapulatus) were captured and sampled at multiple roost locations in the eastern states of Queensland and New South Wales between 2012 and 2014. A range of biological samples (urine and serum, and urogenital, nasal, oral and rectal swabs) were collected from anaesthetized bats, and tested for HeV RNA using a qRT-PCR assay targeting the M gene. Forty-two P. alecto (n = 1410) had HeV RNA detected in at least one sample, and yielded a total of 78 positive samples, at an overall detection rate of 1.76% across all samples tested in this species (78/4436). The rate of detection, and the amount of viral RNA, was highest in urine samples (>serum, packed haemocytes >faecal >nasal >oral), identifying urine as the most plausible source of infection for flying-foxes and for horses. Detection in a urine sample was more efficient than detection in urogenital swabs, identifying the former as the preferred diagnostic sample. The detection of HeV RNA in serum is consistent with haematogenous spread, and with hypothesised latency and recrudesence in flying-foxes. There were no detections in P. poliocephalus (n = 1168 animals; n = 2958 samples) or P. scapulatus (n = 262 animals; n = 985 samples), suggesting (consistent with other recent studies) that these species are epidemiologically less important than P. alecto in HeV infection dynamics. The study is unprecedented in terms of the individual animal approach, the large sample size, and the use of a molecular assay to directly determine infection status. These features provide a high level of confidence in the veracity of our findings, and a sound basis from which to more precisely target equine risk mitigation strategies.

Impact of the invasive rust Puccinia psidii (myrtle rust) on native Myrtaceae in natural ecosystems in Australia

The invasive rust Puccinia psidii (myrtle rust) was detected in Australia in 2010 and is now established along the east coast from southern New South Wales to far north Queensland. Prior to reaching Australia, severe damage from P. psidii was mainly restricted to exotic eucalypt plantations in South America, guava plantations in Brazil, allspice plantations in Jamaica, and exotic Myrtaceous tree species in the USA; the only previous record of widespread damage in native environments is of endangered Eugenia koolauensis in Hawai’i. Using two rainforest tree species as indicators of the impact of P. psidii, we report for the first time severe damage to endemic Myrtaceae in native forests in Australia, after only 4 years’ exposure to P. psidii. A 3-year disease exclusion trial in a natural stand of Rhodamnia rubescens unequivocally showed that repeated, severe infection leads to gradual crown loss and ultimately tree mortality; trees were killed in less than 4 years. Significant (p < 0.001) correlations were found between both incidence (r = 0.36) and severity (r = 0.38) of P. psidii and subsequent crown loss (crown transparency). This provided supporting evidence to conclude a causal association between P. psidii and crown loss and tree mortality in our field assessments of R. rubescens and Rhodomyrtus psidioides across their native range. Assessments revealed high levels of damage by P. psidii to immature leaves, shoots and tree crowns—averaging 76 % (R. rubescens) and 95 % (R. psidioides) crown transparency—as well as tree mortality. For R. psidioides, we saw exceptionally high levels of tree mortality, with over half the trees surveyed dead and 40 % of stands with greater than 50 % tree mortality, including two stands where all trees were dead. Tree mortality was less prevalent for R. rubescens, with only 12 % of trees surveyed dead and two sites with greater than 50 % mortality. Any alternative causal agents for this tree mortality have been discounted. The ecological implications of this are unclear, but our work clearly illustrates the potential for P. psidii to negatively affect Australia’s biodiversity.

Spatiotemporal aspects of Hendra Virus infection in Pteropid Bats (Flying-Foxes) in Eastern Australia

Hendra virus (HeV) causes highly lethal disease in horses and humans in the eastern Australian states of Queensland (QLD) and New South Wales (NSW), with multiple equine cases now reported on an annual basis. Infection and excretion dynamics in pteropid bats (flying-foxes), the recognised natural reservoir, are incompletely understood. We sought to identify key spatial and temporal factors associated with excretion in flying-foxes over a 2300 km latitudinal gradient from northern QLD to southern NSW which encompassed all known equine case locations. The aim was to strengthen knowledge of Hendra virus ecology in flying-foxes to improve spillover risk prediction and exposure risk mitigation strategies, and thus better protect horses and humans. Monthly pooled urine samples were collected from under roosting flying-foxes over a three-year period and screened for HeV RNA by quantitative RT-PCR. A generalised linear model was employed to investigate spatiotemporal associations with HeV detection in 13,968 samples from 27 roosts. There was a non-linear relationship between mean HeV excretion prevalence and five latitudinal regions, with excretion moderate in northern and central QLD, highest in southern QLD/northern NSW, moderate in central NSW, and negligible in southern NSW. Highest HeV positivity occurred where black or spectacled flying-foxes were present; nil or very low positivity rates occurred in exclusive grey-headed flying-fox roosts. Similarly, little red flying-foxes are evidently not a significant source of virus, as their periodic extreme increase in numbers at some roosts was not associated with any concurrent increase in HeV detection. There was a consistent, strong winter seasonality to excretion in the southern QLD/northern NSW and central NSW regions. This new information allows risk management strategies to be refined and targeted, mindful of the potential for spatial risk profiles to shift over time with changes in flying-fox species distribution.

Green and brown bridges between weeds and crops reveal novel Diaporthe species in Australia

Diaporthe (syn. Phomopsis) species are well-known saprobes, endophytes or pathogens on a range of plants. Several species have wide host ranges and multiple species may sometimes colonise the same host species. This study describes eight novel Diaporthe species isolated from live and/or dead tissue from the broad acre crops lupin, maize, mungbean, soybean and sunflower, and associated weed species in Queensland and New South Wales, as well as the environmental weed bitou bush (Chrysanthemoides monilifera subsp. rotundata) in eastern Australia. The new taxa are differentiated on the basis of morphology and DNA sequence analyses based on the nuclear ribosomal internal transcribed spacer region, and part of the translation elongation factor-1α and ß-tubulin genes. The possible agricultural significance of live weeds and crop residues ('green bridges') as well as dead weeds and crop residues ('brown bridges') in aiding survival of the newly described Diaporthe species is discussed.

Queensland Spanner Crab Fishery: Commercial quota setting for June 2016 – May 2018

The Queensland (QLD) fishery for spanner crabs primarily lands live crab for export overseas, with gross landings valued around A$5 million per year. Quota setting rules are used to assess and adjust total allowable harvest (quota) around an agreed target harvest of 1631 t and capped at a maximum of 2000 t. The quota varies based on catch rate indicators from the commercial fishery and a fishery independent survey. Quota management applies only to ‘Managed Area A’ which includes waters between Rockhampton and the New South Wales (NSW) border. This report has been prepared to inform Fisheries Queensland (Department of Agriculture and Fisheries) and stakeholders of catch trends and the estimated quota of spanner crabs in Managed Area A for the forthcoming annual quota periods (1 June 2016–31 May 2018). The quota calculations followed the methodology developed by the crab fishery Scientific Advisory Group (SAG) between November 2007 and March 2008. The QLD total reported spanner crab harvest was 1170 t for the 2015 calendar year. In 2015, a total of 55 vessels were active in the QLD fishery, down from 262 vessels at the fishery’s peak activity in 1994. Recent spanner crab harvests from NSW waters average about 125 t per year, but fell to 80 t in 2014–2015. The spanner crab Managed Area A commercial standardised catch rate averaged 0.818 kg per net-lift in 2015, 22.5% below the target level of 1.043. Compared to 2014, mean catch rates in 2015 were marginally improved south of Fraser Island. The NSW–QLD survey catch rate in 2015 was 20.541 crabs per ground-line, 33% above the target level of 13.972. This represented an increase in survey catch rates of about four crabs per groundline, compared to the 2014 survey. The QLD spanner crab total allowable harvest (quota) was set at 1923 t in the 2012-13 and 2013-14 fishing years, 1777 t in 2014-15 and 1631 t in 2015-16. The results from the current analysis rules indicate that the quota for the next two fishing years be retained at the base quota of 1631 t.

Queensland Spanner Crab Fishery : Commercial quota setting for June 2015 – May 2016

The Australian fishery for spanner crabs is the largest in the world, with the larger Queensland (QLD) sector’s landings primarily exported live overseas and GVP valued ~A$5 million per year. Spanner crabs are unique in that they may live up to 15 years, significantly more than blue swimmer crabs (Portunus armatus) and mud crabs (Scylla serrata), the two other important crab species caught in Queensland. Spanner crabs are caught using a flat net called a dilly, on which the crabs becoming entangled via the swimming legs. Quota setting rules are used to assess and adjust total allowable harvest (quota) around an agreed target harvest of 1631 t and capped at a maximum of 2000 t. The quota varies based on catch rate indicators from the commercial fishery and a fishery-independent survey from the previous two years, compared to target reference points. Quota management applies only to ‘Managed Area A’ which includes waters between Rockhampton and the New South Wales (NSW) border. This report has been prepared to inform Fisheries Queensland (Department of Agriculture and Fisheries) and stakeholders of catch trends and the estimated quota of spanner crabs in Managed Area A for the forthcoming quota period (1 June 2015–31 May 2016). The quota calculations followed the methodology developed by the crab fishery Scientific Advisory Group (SAG) between November 2007 and March 2008. The total reported spanner crab harvest was 917 t for the 2014 calendar year, almost all of which was taken from Managed Area A. In 2014, a total of 59 vessels were active in the QLD fishery, the lowest number since the peak in 1994 of 262 vessels. Recent spanner crab harvests from NSW waters have been about 125 t per year. The spanner crab Managed Area A commercial standardised catch rate averaged 0.739 kg per net-lift in 2014, 24% below the target level of 1.043. Mean catch rates declined in the commercial fishery in 2014, although the magnitude of the decreases was highest in the area north of Fraser Island. The NSW–QLD survey catch rate in 2014 was 16.849 crabs per ground-line, 22% above the target level of 13.972. This represented a decrease in survey catch rates of 0.366 crabs per ground-line, compared to the 2013 survey. The Queensland spanner crab total allowable harvest (quota) was set at 1923 t in 2012 and 2013. In 2014, the quota was calculated at the base level of 1631 t. However, given that the 2012 fisheryindependent survey was not undertaken for financial reasons, stakeholders proposed that the total allowable commercial catch (TACC) be decreased to 1777 t; a level that was halfway between the 2012/13 quota of 1923 t and the recommended base quota of 1631 t. The results from the current analysis indicate that the quota for the 2015-2016 financial year be decreased from 1777 t to the base quota of 1631 t.

An Investigation of Environmental Conditions Experienced During the Life of High Value Wood Components and Products

Australian forest industries have a long history of export trade of a wide range of products from woodchips(for paper manufacturing), sandalwood (essential oils, carving and incense) to high value musical instruments, flooring and outdoor furniture. For the high value group, fluctuating environmental conditions brought on by changes in mperature and relative humidity, can lead to performance problems due to consequential swelling, shrinkage and/or distortion of the wood elements. A survey determined the types of value-added products exported, including species and dimensions packaging used and export markets. Data loggers were installed with shipments to monitor temperature and relative humidity conditions. These data were converted to timber equilibrium moisture content values to provide an indication of the environment that the wood elements would be acclimatising to. The results of the initial survey indicated that primary high value wood export products included guitars, flooring, decking and outdoor furniture. The destination markets were mainly located in the northern hemisphere, particularly the United States of America, China, Hong Kong, Europe including the United Kingdom), Japan, Korea and the Middle East. Other regions importing Australian-made wooden articles were south-east Asia, New Zealand and South Africa. Different timber species have differing rates of swelling and shrinkage, so the types of timber were also recorded during the survey. Results from this work determined that the major species were ash-type eucalypts from south-eastern Australia (commonly referred to in the market as Tasmanian oak), jarrah from Western Australia, spotted gum, hoop pine, white cypress, black butt, brush box and Sydney blue gum from Queensland and New South Wales. The environmental conditions data indicated that microclimates in shipping containers can fluctuate extensively during shipping. Conditions at the time of manufacturing were usually between 10 and 12% equilibrium moisture content, however conditions during shipping could range from 5 (very dry) to 20% (very humid). The packaging systems incorporated were reported to be efficient at protecting the wooden articles from damage during transit. The research highlighted the potential risk for wood components to ‘move’ in response to periods of drier or more humid conditions than those at the time of manufacturing, and the importance of engineering a packaging system that can account for the environmental conditions experienced in shipping containers. Examples of potential dimensional changes in wooden components were calculated based on published unit shrinkage data for key species and the climatic data returned from the logging equipment. The information highlighted the importance of good design to account for possible timber movement during shipping. A timber movement calculator was developed to allow designers to input component species, dimensions, site of manufacture and destination, to see validate their product design. This calculator forms part of the free interactive website www.timbers.com.au.