4 results

Diagnostic detection of aquatic pathogens using real-time next generation sequencing

Project number: 2018-147
Project Status:
Current
Budget expenditure: $216,000.00
Principal Investigator: David Cummins
Organisation: CSIRO Australian Animal Health Laboratory
Project start/end date: 30 Jun 2019 - 28 Oct 2021
Contact:
FRDC

Need

Current diagnostic programs generally rely on highly -specific assays for pathogen detection. While these techniques are invaluable, they are one dimensional and do not provide detailed information critical to a disease investigation. These gaps include the inability to detect unknown pathogens and potential variants of know pathogens and provide no additional genomic or transcriptomic data. Moreover, samples must be shipped to trained personnel in a laboratory, further delaying the time to diagnosis. The MinION, on the other hand, can theoretically detect any pathogen and can potentially be deployed to the field. Moreover, the MinION can rapidly generate full-length genomes, allowing for epidemiological tracking of viral or bacterial strains in near real-time. Such rapid data, which cannot be obtained as quickly using existing methods, are vital if the intention is to intervene in an outbreak and reduce impacts on the productivity and profitability of aquaculture facilities. For example, a rapid, early diagnosis may allow mitigating actions to be taken on-farm, such as the diversion of intake water, movement restrictions of stock and the isolation of infected ponds.
These qualities make the MinION an attractive complimentary platform to fill several gaps in the data obtained during disease outbreak investigations, or routine diagnostics, and potentially for use in the field. However, results from the misuse or lack of understanding of the technology could also have adverse regulatory implications for aquaculture industries. For example, without appropriate guidelines, an inexperienced diagnostician may misinterpret a distant DNA match in a pathogen database as a significant result, this may create unwanted attention to industry and potential stock destruction or changes to disease status that are unjustified. Thus, it is critical that the MinION is evaluated at the Australian Animal Health Laboratory, and guidelines and procedures are developed for accurate diagnostic evaluations. The activities detailed in this application will establish the feasibility of using the MinION for diagnostic applications, and ensure that the data is reliably generated and interpreted appropriately.

Objectives

1. Evaluate if MinION data meets or exceeds the data obtained using established laboratory-based NGS platforms. Objectives (1) and (2) align with Methods section (1).The first objective of this project is to demonstrate if the MinION can obtain quality genome assemblies of known pathogens, such as WSSV, AHPND, OsHV-1 and HaHV that have been created using existing NGS technology. Moreover, determine if the MinION is capable of producing a diagnostic result more rapidly and with greater confidence than traditional techniques. STOP/GO POINT: If MinION data does not produce reliable genome assemblies, no improvement in genome quality, or is significantly more laborious to set-up/run or analyse than existing NGS technologies, do not proceed with objective 2.
2. Evaluate the performance of the MinION using existing diagnostic extraction techniques and produce robust methods and protocols for sample preparation, sequencing and data analysis. This objective will optimise MinION protocols for sample pre-processing, optimal sequencing conditions, and data post-processing. We will then evaluate the MinION data produced from a range of aquatic organisms against data produced using traditional techniques from the same samples. STOP/GO POINT: If after these optimisations, the MinION cannot detect pathogens as reliably as traditional techniques, do not proceed with objective 3.
3. Compare the applicability of MinION to standard molecular assays for identification of pathogens in diagnostic samples. Objective (3) is aligned with Methods section (2).In this objective, diagnostic samples will be tested using existing diagnostics tools (qPCR, cPCR) and MinION sequencing. Analysis between the methods will be detailed, including time to result, pathogen identity and genomic information. This objective will not only provide an insight into real-time sequencing for diagnostics, but in addition the feasibility of MinION technology for field application in the future.

Aquatic Animal Health Technical Forum and Training workshops

Project number: 2018-144
Project Status:
Current
Budget expenditure: $78,000.00
Principal Investigator: David Cummins
Organisation: CSIRO Australian Animal Health Laboratory
Project start/end date: 29 Nov 2018 - 29 Nov 2021
Contact:
FRDC

Need

There is a need to continue the workshops as they provide a forum for representatives from research institutes, Government departments and industry to discuss current aquatic animal health issues facing Australia in a friendly and collaborative setting that encourages open and frank interactions leading to improved mutual understanding of issues facing the different sectors. Workshops have been well supported and have included participants from Government and private laboratories and the aquaculture industry. Industry participants have been from a diversity of farms and representative of a wide variety of aquaculture species. The participant numbers have increased at each workshop from 17 to 35- ideally the maximum group size of 35. This maximum number allows the workshop to be conducted at various locations, as it is not too large a group for host facilities to accommodate.
Previous workshops have led to the exchange of information and methods. This has provided ongoing contacts established at the workshops, for participants to discuss issues, and in turn, respond more effectively to disease outbreaks. With aquaculture facilities and the species farmed continuing to expand in Australia, transferring these vital skills and knowledge to a new generation of researches and those involved in aquatic animal health, will be of benefit to both the aquatic animal health sector and industries.
Due to budgetary constraints in both Government and industry sectors, funding is required to assist workshop participants with travel expenses to attend the workshops. Without partial travel subsidy many forum participants would not obtain authorization to participate. This has repeatedly been raised in feedback and would affect attendance.

The exchange of information and pathways for new people involved in aquatic animal health will be lost and need to be re-established if the workshops fail to continue on an annual basis.

Objectives

1. Source workshop venues, in various States and Territories, that have aquatic animal health capability or services and can accommodate the group size.
2. Organise all aspects of conducting the workshops including
advertising through Health Highlights subscription and peer referrals, guest presenter, presentation program, field trips and practical sessions, accommodation and catering.
3. Encourage new and emerging science and production staff to attend the AAHTF and to gain experience in making presentations
4. Continue to update the contact list and email distribution list/group for continued information exchange
5. Reports and financial acquittals prepared according to milestone schedule.
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