5 results
Environment
PROJECT NUMBER • 2013-004
PROJECT STATUS:
COMPLETED

Aquatic Animal Health Subprogram: the Neptune Project- a comprehensive database of Australian aquatic animal pathogens and diseases

Aquatic animal health experts from the Queensland Museum (QM) have been completing work on a parasite and disease database called Neptune. Work on Neptune has taken place at QM in Brisbane since May 2013, resulting in the completion of major improvements to the database. These will allow Neptune to...
ORGANISATION:
Queensland Museum
TAGS
Software
Policy
Laboratory Procedures
Disease
Data

Aquatic Animal Health Subprogram: identification of host interactions in the life-cycle of QX disease

Project number: 2006-062
Project Status:
Completed
Budget expenditure: $88,874.24
Principal Investigator: Rob D. Adlard
Organisation: Queensland Museum
Project start/end date: 30 Aug 2006 - 30 Jun 2008
Contact:
FRDC
TAGS
Supply Chain
Safety
Food Safety
Consumer
Analytics
SPECIES
Sydney Rock Oyster

Need

There have been some significant advances in our knowledge of QX disease of Sydney rock oysters in recent years. The pathogen has been isolated from many farming areas without being accompanied by patent disease and the influence of host fitness together with environmental effectors are now being implicated as disease precursors. Nonetheless, the devastating oyster mortalities in the Hawkesbury River this year (2005) highlight our problems in devising intelligent management strategies to minimise the impact of this disease.
A key obstacle to developing knowledge on parameters that control whether a disease outbreak will occur or whether the pathogen remains in estuaries at or beneath detectable levels is the lack of an experimental model of infection. In turn, the development of such a model is predicated on identifying the alternate (i.e. intermediate) host in the lifecycle of QX disease, a stage which is required for the pathogen to viably cycle repeatedly through an estuary. Furthermore, if an experimental model could be identified and later developed, obvious benefit would flow to strategic programs of selective breeding for disease resistant oysters. For example, an experimental model of infection would then provide a consistent and quantifiable challenge to assess the level of resistance in selected stock. Equally, the interactions of oyster immuno-competence and environment could then be assessed in a controlled system without the risk of spatial and temporal variation in QX disease prevalence and intensity that occur in natural estuarine systems.
A successful outcome of this research would have major benefit to our understanding of the biology of QX disease and have direct application to parallel projects aimed at benefitting the industry in both Queensland and New South Wales.

Objectives

1. To determine what members of the macrofauna contribute as intermediate hosts in the life-cycle of Marteilia sydneyi, agent of QX disease.
2. To identify and characterise previously unknown stages of Marteilia sydneyi through in-situ DNA probe hybridisation and histological examination.

Final report

Author: Robert Adlard
Final Report • 2017-09-29 • 5.52 MB
2006-062-DLD.pdf

Aquatic Animal Health Subprogram: validation of DNA-based (PCR) diagnostic tests suitable for use in surveillance programs for marteiliosis of rock oysters in Australia

Project number: 2001-630
Project Status:
Completed
Budget expenditure: $58,279.00
Principal Investigator: Rob D. Adlard
Organisation: Queensland Museum
Project start/end date: 30 Jan 2002 - 30 Jun 2005
Contact:
FRDC
TAGS
Training
Trade
Fisheries Management
Disease
Biosecurity
SPECIES
Sydney Rock Oyster

Need

Marteiliosis (QX disease, aetiological agent the protozoan parasite marteilia sydneyi) typically causes serious, seasonally recurrent mortalities in farmed and wild rock oysters in eastern Australia. The disease is listed as notifiable by the OIE and is included on the Australian National List of Reportable Disease of Aquatic Animals.

The OIE has recently adopted the concept of zoning to facilitate trade and to prevent spread of disease within a country. In turn, Australia has recognised the value of zoning in its aquaculture industries with the adoption and endorsement of Zoning Policy Guidelines by Standing Committee on Fisheries and Aquaculture.

The establishment of scientifically defensible zoning and translocation policies, particularly in relation to QX disease control, is critical to the long term development of the rock oyster aquaculture industry. NSW Fisheries currently prohibits movement of oysters from known QX infected estuaries to those thought to be free of infection. However, given the many millions of rock oysters translocated annually between NSW estuaries of undetermined disease status, there is an urgent need to accurately identify free an infected zones. This, in turn, depends upon the availability of standardised, validated diagnostic tests.

Histopathology is currently viewed as the 'gold standard' for QX disease diagnosis, while preliminary comparative data (Callinan and Wesche, unpublished data) suggest that an alternative cytological method, stained tissue imprints of oyster digestive gland, has a sensitivity of 60% and specifically of 100%. Recently, however, there have been major advances in development of PCR tests for marteiliosis (Berthe et al. 2000; Kleeman and Adlard 2000). It is possible that PCR can be used to confirm presumptive/inconclusive diagnoses obtained by histopathology or cytology. PCR may also have potential as a cheap and reliable mass screening diagnostic test. In either event, however, rigorous standardisation and validation will be necessary before a PCR test can be accepted for use in zoning-related QX disease surveillance.

Objectives

1. Production of a fully validated, standard PCR diagnostic test for the presence of marteilia sydneyi in oyster tissue capable of identifying marteilia sydneyi to species level and with a high level of sensitivity.
2. Assessment of comparative cost/benefit of histological, cytological and PCR diagnostic methods for identification of marteilia sydneyi.
3. Production of an Australian and New Zealand Standard Diagnostic Procedure (ANZSDP) for marteiliosis.

Final report

ISBN: 0-9751116-1-2
Author: Robert Adlard
Final Report • 2003-11-18 • 350.59 KB
2001-630-DLD.pdf

Aquatic Animal Health Subprogram: development of a disease zoning policy for marteiliosis to support sustainable production, health certification and trade in the Sydney rock oyster

Project number: 2001-214
Project Status:
Completed
Budget expenditure: $281,226.02
Principal Investigator: Rob D. Adlard
Organisation: Queensland Museum
Project start/end date: 6 Jun 2001 - 15 Jul 2005
Contact:
FRDC
TAGS
Trade
Sustainability
Disease
Diagnostic
Aquaculture
SPECIES
Sydney Rock Oyster

Need

The rock oyster industry in Australia is currently valued at around $28 million annually. The current output is about half of the industry peak in the late 1970’s. For the industry to survive in the long-term requires the ability to service what may become a premium domestic market demanding a high quality product. The expansion of the industry is likely to be available only from international export, which in turn requires compliance with international regulations on oyster health with a transparent health audit trail. The rock oyster is potentially positioned for re-emerging export success, being a unique product with an extended shelf-life relative to other oyster species (e.g. the Pacific oyster, Crassostrea gigas) and this is an opportunity that should be exploited by the industry.

The techniques of surveillance and diagnosis for molluscan pathogens required by the OIE for imported oyster products are not only stringent and accepted as the worldwide standard, but are also applicable to domestic requirements within Australia. In essence, the regulations state that appropriate diagnostic tests are applied for detecting the presence of pathogens of molluscs (microscopic identification techniques with the potential for specific molecular identification using monoclonal antibodies or DNA probes) which have been collected as part of a surveillance program within delimited coastal zones. The sample size, period and frequency are determined with reference to the cycle of infection of the particular pathogen and its prepatent period. There is an initial 2 year period of surveillance before a zone can be granted a disease-free status, with ongoing surveillance required for this status to be maintained.

The development of a zoning policy framework for marteiliosis will provide a valuable opportunity to implement and field-test Australia’s zoning policy guidelines in a practical context to assist with the development of further zoning policies for diseases of aquatic animals. Considerable interest has already been expressed in the case study by State authorities and it will be discussed at an Aquatic Animal Disease Zoning Workshop in Canberra on 23 January 2001, hosted by the National Offices of Animal and Plant Health. Furthermore, the development of the zoning policy will be of direct benefit to the oyster industry by facilitating domestic and international market access, and through identifying and protecting the remaining disease-free production areas

Objectives

1. 1. The primary objective is to implement and field-test the zoning policy framework developed under Aquaplan in a practical context and to facilitate the development of further zoning policies for other significant diseases of aquatic animals. This will be conducted using marteiliosis as a case study to develop an effective zoning policy that is consistent with internationally recognised (OIE) standards. The zoning policy will aim to:* Reduce the risk of introducing this pathogen into the remaining disease-free production areas
and* Facilitate domestic and international market access for the industry.
2. 2. The sub-objectives necessary to achieve this are to:* Identify through sampling and appropriate diagnosis marteiliosis-free and marteiliosis-endemic estuaries within oyster culture areas
* Determine the specific identity of Marteilia sp. from positive samples through ultra-structural and molecular diagnostics
* Develop a rational and effective program of surveillance for marteiliosis, based on occurrence and an assessment of risk for each oyster producing estuary
* In consultation with fisheries managers and industry, develop a coastal zoning plan for marteiliosis.

Final report

ISBN: 0-9751116-3-9
Author: Robert Adlard
Final Report • 2006-02-01 • 975.12 KB
2001-214-DLD.pdf

Summary

The edible oyster industry in Australia is currently valued at around $62.5 million annually of which rock oyster production accounts for approx 56%. For the industry to survive in the long-term requires the ability to service what may become a premium domestic market demanding a high quality product.  The expansion of the industry is likely to be available only from international export, which in turn requires compliance with international regulations on oyster health with a transparent health audit trail.  The rock oyster is potentially positioned for re-emerging export success, being a unique product with an extended shelf-life relative to other oyster species (e.g. the Pacific oyster, Crassostrea gigas) and this is an opportunity that should be exploited by the industry.
 
Within Australia, the Sydney Rock Oyster industry is subjected to periodic epizootics of disease induced by a range of parasitic organisms that produce significant mortality and morbidity of commercial oyster stocks.  The most significant of these is the agent responsible for ‘QX disease’ (caused by the protistan parasite Marteilia sydneyi) affecting the Sydney rock oyster Saccostrea glomerata.  Management of this disease has been based on quarantine of affected estuaries enforced through limitation on the movement of potentially infected stock.  In this context, it was obvious that the oyster industry required a disease zoning policy based on scientifically defensible data to allow domestic best practice in oyster farming and to maximise market accessibility for the industry.  This host/parasite system then formed the basis for a test of the zoning policy framework developed under the federal government’s ‘AQUAPLAN’.
 
A number of key issues related to zoning and surveillance for specific diseases were addressed through this project.  Initially the design of field collection and the appropriate test to use for diagnosis were assessed to maximise, and allow quantification of, disease detection limits in the surveillance program.
 
1. The design of field sampling to identify disease infected oysters was critical in order to reach a statistically robust probability of disease detection.  Global animal health standards (Office Internationale des Epizooties) recommend random sampling from a zone to detect a prevalence of 2% or greater disease in a population.  This was fulfilled using a computer generated random selection of geographic co-ordinates under which individual oysters were sampled (Angus Cameron, AusVet).
 
2. The appropriate method for diagnosis of disease, another critical issue in disease surveillance programs, was assessed by comparing the sensitivity and specificity of: tissue imprints (cytology); or tissue sections (histology); or the presence of specific parasite DNA (by polymerase chain reaction - PCR).  Our analysis showed clearly that PCR was the most sensitive diagnostic test followed by cytology then histology.  PCR also detected the presence of sub-clinical infections which could not be unambiguously identified using either histology or cytology.  Confirmatory diagnosis (following PCR) at sub-clinical levels was undertaken using DNA in situ hybridisation tests designed to stain the QX organism specifically in tissue section.
 
Combined surveillance results from 2001 (NSW estuaries only), 2002-03 (NSW and Queensland estuaries) and 2004 (Queensland estuaries only) demonstrated some significant departures from the geographic distribution expected for QX disease.  In 2001 diagnosis was undertaken using cytology and no unexpected occurrences of the disease were observed, with positives recorded only from the Clarence River (1.5% of sample infected), Georges River (47% of sample infected).  In 2002 the distribution of disease was significantly different to that expected.  Initially using cytology for diagnosis there were no apparent unusual infections with Southern Moreton Bay (0.8% of sample infected), Richmond River (40.8% of sample infected), Clarence River (22% of sample infected) and Georges River (16% of sample infected) recording oysters positive for the disease.  However, when PCR techniques were used for diagnosis in estuaries that had never recorded the presence of the disease agent it became obvious that the organism was more widespread than indicated by previous diagnostic testing or previous occurrences of disease outbreaks.  In total 142 unexpected positives for Marteilia sydneyi were found in oysters scored as negative by cytological examination during surveillance in this project.  Of these, 61 were identified in oysters sampled from estuaries with no prior record of Marteilia sydneyi.  These represent oysters from Hastings River, Wallis Lake, Port Stephens, Bateman’s Bay, Tuross Lake, Narooma and Merimbula.
 
Further testing of these infections confirmed the identity of the QX organism and found it to be present in the oyster tissues at a sub-clinical level i.e. prior to reaching the oyster’s digestive gland where the parasite would normally produce spores.  At this stage of development, pathology in the oyster is reduced and the condition factor of oysters is not seriously compromised.
 
In 2003 surveillance and diagnosis using PCR techniques showed a reduced impact of QX disease with Southern Moreton Bay (0.67% of sample infected), Brunswick River (1.3% of sample infected), Richmond River (13.3% of sample infected), Clarence River (6% of sample infected) and Georges River
(0.67% of sample infected).
 
This project has had a significant impact on our understanding of QX disease in rock oysters as it applies to management.  Rather than the disease agent being limited geographically to those estuaries that experience periodic outbreaks, the agent has been identified in most rock oyster growing areas on the east coast of Australia.  As such there is the potential for outbreaks of QX disease in all commercial growing areas (indeed such an outbreak occurred in 2004, with seasonal re-occurrence in 2005, in the Hawkesbury River) and that disease is likely to be regulated through a combination of the dynamics of the parasite lifecycle and the level of oyster fitness.  Furthermore, in any aquatic system the environment will play an equally significant role in the outcomes of host/parasite interactions both through direct impact on stages (spores, infective stages) in the lifecycle of the parasite and indirectly through its impact on host fitness.
 
In the light of our new understanding of the distribution of the QX disease agent it could be argued that management through quarantine of identified QX-endemic estuaries is no longer appropriate.  However, the biology of Marteilia sydneyi (dynamics of the life cycle of the parasite, interactions with alternate hosts) and its interaction with the host oyster’s immune system are incompletely understood and the precautionary principle should be upheld especially in the case of such a serious disease.
 
While estuaries which undergo periodic outbreak should remain closed to export of oysters for relaying live in water elsewhere, local management will focus on disease seasonality and stock rotation to avoid the high risk periods in mid to late summer.  These periods should be identified with accuracy to maximise available growth periods in disease endemic areas of estuaries.  The ongoing projects to develop QX disease resistant oysters (NSW DPI and collaboration with Macquarie University) should run parallel with a program of incremental addition to the biological knowledge of this pathogen.  Specifically, an absence of our ability to maintain a laboratory based infection model hampers research on identifying those factors (pathogen-specific, oyster-specific and environment-specific) which promote disease.
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