- Animals: shellfish
- Pest Management: disease vectors
We propose to develop and commercialize loop-mediated isothermal amplification (LAMP) assays for farm-based detection of six virulence marker genes from four Vibrio species associated with shellfish hatchery failures, along with an easy-to-understand Hatchery Instructions Guide for the assays. This proposal has been developed in response to an increased number of shellfish hatchery failures, which ultimately impact hundreds of oysters farms in the Northeast who depend on hatcheries for seed. Vibriosis, infection of larval or juvenile oysters by pathogenic Vibrio spp. is frequently a cause of these failures, reducing dependability of seed production. From a survey of hatchery managers, 86.7% indicated they would adopt the use of this assay into their hatchery operations to help diagnose production problems or verify that no pathogens are present in the algae they are growing as food for the larvae and seed. LAMP assays will be developed at Bigelow Laboratory for Ocean Sciences in Year 1 of the project, resulting in rapid, inexpensive molecular assays that can be implemented directly by hatchery operators, instead of sending samples out for expensive, long-turnaround PCR assays. The quick return of presence/absence of pathogens will let hatchery operators quickly develop management plans and mitigation strategies to remove the pathogens. The developed assays will be tested in the Mook Sea Farm (MSF) Hatchery in Maine, to verify that the protocols are appropriate for hatchery use. The MSF team will develop clear protocols and supplies lists in the Hatchery Instructions Guide, which will be shared with the Project Advisory Committee and a Hatchery Feedback Group to ensure the guide is user-friendly for those without molecular biology backgrounds. MSF and Bigelow Laboratory will seek commercialization opportunities for the custom LAMP assays, including but not limited to ‘kit production’, to ensure that high-quality LAMP assays are readily available to end users in a convenient, reliable, and contamination-free ‘easy to use’ format. Lessons learned through the development of these hatchery-focused LAMP assays will simplify the steps needed to develop LAMP assays for additional pathogens that are of concern to shellfish hatcheries and grow-out operations.
Performance targets from proposal:
We propose to develop and commercialize loop-mediated isothermal amplification (LAMP) assays for farm-based detection of six virulence marker genes from four Vibrio species associated with shellfish hatchery failures, along with an easy-to-understand Hatchery Instructions Guide. The LAMP method allows for rapid, inexpensive, colorimetric detection of pathogens with minimal sample preparation, giving shellfish hatcheries straightforward diagnostic tools to manage hatchery production. These tools can be used to detect infections and determine management strategies, as well as to check shellfish seed viability prior to shipping to farms from the hatcheries, giving confidence to those purchasing the seed for their farms.