FY07-09 proposal 200729400
Jump to Reviews and Recommendations
Section 1. Administrative
| Proposal title | Control of BKD by Inactivation of the Renibacterium salmoninarum Sortase Enzyme as an Alternative to Antibiotics |
| Proposal ID | 200729400 |
| Organization | Northwest Fisheries Science Center |
| Short description | This proposal describes research to determine if the sortase gene product of Renibacterium salmoninarum can be a target for therapeutic drugs against Bacterial Kidney Disease of salmon. |
| Information transfer | Salient findings of the project will be presented at national and international symposia and workshops and communicated to various stakeholders of the project. Results also will be made available to the public by publishing in peer reviewed international journals. If a suitable drug with anti-BKD activity is identified during this study, further detailed investigation of the drug will be carried out for generating sufficient data required for getting the drug approved by FDA for BKD treatment. |
| Proposal contact person or principal investigator | |
Contacts
| Contact | Organization | |
|---|---|---|
| Form submitter | ||
| Mark Strom | Northwest Fisheries Science Center/NOAA Fisheries | mark.strom@noaa.gov |
| All assigned contacts | ||
| Mark Strom | Northwest Fisheries Science Center/NOAA Fisheries | mark.strom@noaa.gov |
Section 2. Locations
Province / subbasin: Mainstem/Systemwide / Systemwide
| Latitude | Longitude | Waterbody | Description |
|---|---|---|---|
| 47/39 | 122/18 | Northwest Fisheries Science Center, Montlake Laboratory | |
| 47/34 | 122/33 | Northwest Fisheries Science Center, Manchester Marine Laboratory |
Section 3. Focal species
primary: All Anadromous Salmonidsprimary: Chinook All Populations
primary: Sockeye All Populations
secondary: Coho Unspecified Population
secondary: Steelhead All Populations
Section 4. Past accomplishments
| Year | Accomplishments |
|---|
Section 5. Relationships to other projects
| Funding source | Related ID | Related title | Relationship |
|---|---|---|---|
| BPA | 199204000 | Redfish Lake Sockeye Broodstoc | prevention of BKD |
| BPA | 199606700 | Manchester Spring Chinook Capt | prevention of BKD |
| BPA | 199305600 | Demonstration of Captive Salmo | Carried out research within this project to test antibiotic therapies and vaccination strategies to control BKD |
| [Funding Source left blank] | [no entry] | Genome sequencing of the vertically-transmitted fish pathogen Renibacterium salmoninarum | USDA/NSF funded program (PI Strom is the PI of this program) to sequence the R. salmoninarum genome. Discoveries from the genome project are directly related to the current proposal. |
Section 6. Biological objectives
| Biological objectives | Full description | Associated subbasin plan | Strategy |
|---|---|---|---|
| Determine if anti-sortase treatment prevents BKD | To investigate the pathogenicity of virulence attenuated R. salmoninarum (Rs) in Chinook salmon and the suitability of using vinyl sulfone drugs as treatment options for BKD. | None | Determine virulence of Rs sortase mutants in Chinook salmon; determine toxicity of vinyl sulfones to CHSE-214 cells and Chinook salmon; determine efficacy of vinyl sulfone treatment of BKD in Chinook salmon |
| Determine if Rs sortase is involved in virulence | To investigate if deletion of the sortase gene of Renibacterium salmoninarum (Rs), or inhibition of sortase activity by vinyl sulfones, prevents cell surface display of important virulence-associated proteins and to assess the ability of these bacteria to adhere and grow in fish cells. | None | Construction of sortase mutants and analysis of ability of bacteria to adhere to eukaryotic (host) cell fibronectin and cells; determine if vinyl sulfones can inhibit Rs sortase activity; determine if sortase inhibition prevents intracellular growth of Rs |
Section 7. Work elements (coming back to this)
| Work element name | Work element title | Description | Start date | End date | Est budget |
|---|---|---|---|---|---|
| Collect/Generate/Validate Field and Lab Data | Construction of sortase mutants. | Construct mutants of R. salmoninarum deficient in expression of the sortase gene. | 6/1/2007 | 1/1/2008 | $60,094 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Analyze/Interpret Data | Analysis of R. salmoninarum sortase mutants and confirmation of gene inactivation. | Verify sortase gene mutation and assess ability of wild type and mutant R. salmoninarum to bind to fibronectin. | 9/1/2007 | 1/1/2008 | $50,829 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Collect/Generate/Validate Field and Lab Data | Vinyl sulfone inhibition of sortase activity. | Test the ability of a number of vinyl sulfones to inhibit R. salmoninarum sortase activity. | 10/1/2007 | 3/1/2008 | $48,075 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Analyze/Interpret Data | Selection of drug with maximum sortase inhibitory activity. | Perform statistical analysis on sortase inhibitory activity to select those with most potent inhibitory activity. | 10/1/2007 | 3/1/2008 | $40,663 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Collect/Generate/Validate Field and Lab Data | Characterization and identification of sortase-mediated cell surface anchored proteins. | Carry out a differential proteomic analysis of membrane proteins between srtA+ and strA- mutants or bacteria treated with vinyl sulfones to identify sortase mediated surface anchored proteins. | 3/1/2008 | 8/1/2008 | $12,019 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Analyze/Interpret Data | Analysis of proteomic data and identification of surface-bound proteins. | Carry out LC MS/MS analysis of peptides to identify surface-bound proteins localized through the sortase. | 3/1/2008 | 8/1/2008 | $10,166 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Collect/Generate/Validate Field and Lab Data | Ability of of srtA- mutants to adhere, grow and survive in CHSE-214 cells compared to wild type R. salmoninarum. | Determine the role of sortase and sortase mediated surface proteins in the growth and survival of R. salmoninarum in Chinook salmon cells. | 8/1/2008 | 1/1/2009 | $58,280 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
||||
| Analyze/Interpret Data | Analysis of cell adherence and intracellular growth data. | Perform statistical analysis of the data to determine the role of sortase in the ability of R. salmoninarum to adhere to fish cells and grow intracellularly. | 8/1/2008 | 1/1/2009 | $43,484 |
|
Biological objectives Determine if Rs sortase is involved in virulence |
Metrics Focal Area: Systemwide Secondary R, M, and E Type: Uncertainties Research |
||||
| Collect/Generate/Validate Field and Lab Data | Virulence of sortase mutants to Chinook salmon. | Demonstrate that the deletion of the sortase gene and the resultant attenuation of virulence is manifested as reduced severity of BKD and reduced mortality in Chinook salmon challenge models. | 1/1/2009 | 6/1/2009 | $58,280 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Analyze/Interpret Data | Analysis of challenge data and determination of virulence of sortase mutants. | Perform statistical analysis of fish challenge data comparing wild type to the sortase mutant of R.salmoninarum. | 1/1/2009 | 6/1/2009 | $43,484 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
||||
| Collect/Generate/Validate Field and Lab Data | Toxicity of vinyl sulfones to CHSE-214 cells and Chinook salmon. | Determine toxicity of vinyl sylfones to CHSE-214 cells and Chinook salmon to assess their suitability as antivirulence chemotherapeutics in fish. | 6/1/2009 | 11/1/2009 | $61,766 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
||||
| Analyze/Interpret Data | Analysis of drug toxicity data. | Perform statistical analysis to assess toxicity of vinyl sulfones to fish cells and chinook salmon. | 6/1/2009 | 11/1/2009 | $35,098 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
||||
| Collect/Generate/Validate Field and Lab Data | Effect of vinyl sulfones in attenuating the virulence of R. salmoninarum in Chinook salmon. | Test most effective vinyl sulfone(s) on Chinook salmon challenged with low and high doses of R. salmoninarum. | 11/1/2009 | 5/31/2010 | $61,766 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
||||
| Analyze/Interpret Data | Analysis of data on the therapeutic effect of drugs on fish and fish cells. | Perform statistical analysis to determine the therapeutic effect of vinyl sulfones on R. salmoninarum challenged chinook salmon. | 11/1/2009 | 5/31/2010 | $35,098 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics Focal Area: Systemwide Primary R, M, and E Type: Uncertainties Research |
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| Produce Hatchery Fish | Culture Chinook salmon for challenge, toxicity, and drug treatment experiments. | George Adams stock fall Chinook salmon will be obtained and cultured for all challenge, toxicity, and drug treatment experiments. Fishes of ~10 – 50 g size will be used for all experiments. Fish will be reared at the University of Washington’s Big Beef Creek facility on well water (10°C), at the Manchester Research Station on filtered, UV-treated seawater, or at the Mukilteo Research Station on filtered, UV-treated seawater. All experimentally infected fish will be autoclaved prior to disposal. | 1/1/2009 | 5/31/2010 | $89,257 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD |
Metrics * Rearing: # juveniles (presmolt) into program: fish ponded |
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| Coordination | Project planning and coordination and communication of results. | The elements under Objectives 1 and 2 will be coordinated by the Principal Investigator. Salient findings of the project will be presented at national and international symposia/ workshops and communicated to various stakeholders of the project. Results also will be made available to the public by publishing in peer reviewed international journals. | 6/1/2007 | 5/31/2010 | $5,569 |
|
Biological objectives Determine if anti-sortase treatment prevents BKD Determine if Rs sortase is involved in virulence |
Metrics |
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Section 8. Budgets
Itemized estimated budget
| Item | Note | FY07 | FY08 | FY09 |
|---|---|---|---|---|
| Personnel | [blank] | $100,194 | $107,783 | $112,961 |
| Fringe Benefits | [blank] | $44,670 | $51,241 | $53,113 |
| Supplies | [blank] | $22,000 | $19,500 | $22,800 |
| Travel | [blank] | $2,904 | $2,904 | $2,904 |
| Capital Equipment | [blank] | $0 | $0 | $0 |
| Overhead | [blank] | $45,995 | $49,516 | $51,650 |
| Other | water charges for fish rearing, rents and utilities for fish rearing | $7,931 | $7,931 | $7,931 |
| Totals | $223,694 | $238,875 | $251,359 | |
Total estimated FY 2007-2009 budgets
| Total itemized budget: | $713,928 |
| Total work element budget: | $713,928 |
Cost sharing
| Funding source/org | Item or service provided | FY 07 est value ($) | FY 08 est value ($) | FY 09 est value ($) | Cash or in-kind? | Status |
|---|---|---|---|---|---|---|
| Totals | $0 | $0 | $0 | |||
Section 9. Project future
|
FY 2010 estimated budget: $0 FY 2011 estimated budget: $0 |
Comments: [Outyear comment field left blank] |
Future O&M costs: There are no future operations and maintenance costs. If promising results are obtained showing that BKD can be treated by small molecule sulfones to inhibit virulence, It is anticipated that a new proposal would be submitted in FY 2010 to move the treatment into wide spread clinical trials.
Termination date: 5/31/2010
Comments: It is expected that the objectives outlined in the proposal will be achieved by the end of the project, 5/31/2010. Work in future years may include carrying out pharmacokinetics of vinyl sulfones in fish, and applications to other Gram-positive pathogens of fish, such as Streptococcus iniae.
Final deliverables: Results and data will be made available through meeting presentations, contract reports, and publication of manuscripts in peer-reviewed journals. If vinyl sulfones are efficaceous in treatment of BKD, the technology and methodology will be readily available to various stakeholders (hatchery managers, fish health professionals, etc.).
Section 10. Narrative and other documents
Reviews and recommendations
| FY07 budget | FY08 budget | FY09 budget | Total budget | Type | Category | Recommendation |
|---|---|---|---|---|---|---|
| NPCC FINAL FUNDING RECOMMENDATIONS (Oct 23, 2006) [full Council recs] | ||||||
| $0 | $0 | $0 | $0 | Expense | Basinwide | Do Not Fund |
| NPCC DRAFT FUNDING RECOMMENDATIONS (Sep 15, 2006) [full Council recs] | ||||||
| $0 | $0 | $0 | $0 | Basinwide | ||
ISRP PRELIMINARY REVIEW (Jun 2, 2006)
Recommendation: Fundable
NPCC comments: This is an excellent proposal, with excellent technical and scientific background. Potential benefits to fish are clear. In reality, many hatchery operations (and the populations they impact) could be affected positively. Project proponents have a good track record of peer reviewed publications and participation in fisheries meetings. Results of the work could have significant and lasting benefits to the artificial production programs, including reducing the impact and risks of these programs to natural fish populations.
ISRP FINAL REVIEW (Aug 31, 2006)
Recommendation: Fundable
NPCC comments: This is an excellent proposal, with excellent technical and scientific background. Potential benefits to fish are clear. In reality, many hatchery operations (and the populations they impact) could be affected positively. Project proponents have a good track record of peer reviewed publications and participation in fisheries meetings. Results of the work could have significant and lasting benefits to the artificial production programs, including reducing the impact and risks of these programs to natural fish populations.