Spatial distribution of mortality in Pacific oysters Crassostrea gigas: reflection on mechanisms of OsHV-1 transmission

The ostreid herpesvirus (OsHV-1) has the potential to devastate Crassostrea gigas culture in Australia as it has done in many other countries, highlighting the need for a better understanding of disease expression and transmission. The aim of this study was to assess the spatial distribution of OsHV-1 associated mortalities in one of only two infected areas in Australia, Woolooware Bay (Botany Bay New South Wales). In October 2011, healthy sentinel Pacific oysters were placed in three different locations at three different tidal levels and OsHV-1 associated mortalities were closely monitored over 7 months. The outbreak started in November 2011 and the disease remained active until April 2012. Three major mortality events were detected. Rather than being a propagating epizootic, it appeared that most oysters were infected from the same environmental source. The distribution of OsHV-1 associated mortalities was spatially clustered, highly variable and clearly dependent on the age of oysters and their position in the water column. Non-random distribution of mortalities at macro scale (sites several km apart) and micro scale (within rearing trays), and vertical clustering patterns in the water column are discussed in relation to mechanisms of transmission in water. We hypothesise that OsHV-1 may be carried through water by particles, possibly plankton. Key words: Crassostrea gigas, Ostreid herpesvirus 1, summer mortalities, spatial distribution, plankton, disease transmissionFunded by the Fisheries Research and Development Corporation, the University of Sydney and the Sydney Metropolitan Catchment Management Authorit

A Search for Distant Galactic Cepheids Toward l=60

We present results of a survey of a 6-square-degree region near l=60, b=0 to search for distant Milky Way Cepheids. Few MW Cepheids are known at distances >~ R_0, limiting large-scale MW disk models derived from Cepheid kinematics; this work was designed to find a sample of distant Cepheids for use in such models. The survey was conducted in the V and I bands over 8 epochs, to a limiting I~=18, with a total of ~ 5 million photometric observations of ~ 1 million stars. We present a catalog of 578 high-amplitude variables discovered in this field. Cepheid candidates were selected from this catalog on the basis of variability and color change, and observed again the following season. We confirm 10 of these candidates as Cepheids with periods from 4 to 8 days, most at distances > 3 kpc. Many of the Cepheids are heavily reddened by intervening dust, some with implied extinction A_V > 10 mag. With a future addition of infrared photometry and radial velocities, these stars alone can provide a constraint on R_0 to 8%, and in conjunction with other known Cepheids should provide good estimates of the global disk potential ellipticity.Comment: 18 pages, 4 tables, 13 figures (LaTeX / AASTeX

Experimental infections of Pacific oyster Crassostrea gigas using the Australian OsHV-1 µVar strain

In Australia the spread of the Ostreid herpesvirus OsHV-1 microvariant (OsHV-1 µVar) threatens the Pacific oyster industry. There was an urgent need to develop an experimental infection model in order to study the pathogenesis of the virus in controlled laboratory conditions. The present study constitutes the first attempt to use archived frozen oysters as a source of inoculum, based on the Australian OsHV-1 µVar strain. Experiments were conducted to test (i) the virus infectivity, (ii) the dose-response relationship for OsHV-1, and (iii) the best storage conditions to keep infective viral inoculum. Intramuscular injection of a viral inoculum consistently led to an onset of mortality 48h post injection and a final cumulative mortality exceeding 90%, in association with high viral loads (1 x 105 – 3 x 107 copies of virus mg-1) in dead individuals. For the first time an infective inoculum was produced from frozen oysters (tissues stored at -80°C for 6 months). Storage of purified viral inoculum at +4°C for 3 months provided similar results to use of fresh inoculum whereas storage at -20°C, -80°C and room temperature was detrimental to infectivity. A dose-response relationship for OsHV-1 was identified but further research is recommended to determine the most appropriate viral concentration for development of infection models that would be used for different purposes. Overall, this work highlights the best practices and potential issues that may occur in the development of a reproducible and transferable infection model for studying the pathogenicity of the Australian OsHV-1 strain in C. gigas under experimental conditions. Keywords: Ostreid herpesvirus 1; Crassostrea gigas; experimental infection model; dose-response; inoculum preparation; storage condition

Descriptive epidemiology of mass mortality due to Ostreid herpesvirus-1 (OsHV-1) in commercially farmed Pacific oysters (Crassostrea gigas) in the Hawkesbury River estuary, Australia

postprintMortality of farmed triploid Pacific oysters (Crassostrea gigas) associated with Ostreid herpesvirus-1 (OsHV-1) was first recorded in Australia in the Georges River/Botany Bay estuary (New South Wales) in late 2010. Two years later, the first sign of possible inter-estuarine spread was observed when commercial triploid Pacific oysters in the Hawkesbury River estuary, located 50 km north of Botany Bay, were affected by mass mortality. The aim of this study was to describe the epidemiological features of the Hawkesbury outbreak via a formal investigation which was conducted in real time and comprised: an assessment of stock levels, past oyster acquisitions and a trace forward investigation to identify stock at greatest risk due to transfers of oysters; passive surveillance of the spread of mortalities in the estuary; active surveillance using PCR to identify the distribution of OsHV-1 infection on farms in the estuary and mortality estimates to identify age and size classes affected; identification of the time of first infection using data from sentinel oysters; and assessment of environmental risk factors. Mortalities were recorded in all age classes but were greater in spat and juveniles than in adults. The incubation period for mass mortality was < 4 days, however subclinical OsHV-1 infection was detected three months prior to the first signs of mortality in the index case (first location affected), which suggests that low viral loads of OsHV-1 are insufficient to induce the disease. While inefficient oyster-to-oyster transmission occurred at two locations, a synchronous infection arising from a common environmental source was required to explain the mass mortalities at the index case (Mullet Creek). Estuarine hydrodynamics then assisted rapid dispersal of viral particles throughout the estuary. Seawater temperatures were consistently above 24°C during the month preceding mass mortalities with variations (± 3°C) being observed over a few days during this period; however this did not necessarily lead to mortality events when the virus was present. There was no evidence of prior movement of potentially infected oysters or farming equipment into the Hawkesbury River estuary to explain the outbreak. Key words: Disease outbreak investigation; Pacific oysters; Ostreid herpesvirus-1; disease transmission; incubation period; water temperature; epidemiologyFunded by the Fisheries Research Development and Corporation (FRDC), The University of Sydney and the Sydney Metropolitan Catchment Management Authorit