FOCUS AREA RFP. Project Objective

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1 This project is being funded through the Focus Area Program, which enables WRF to solve broadly relevant subscriber issues and challenges with a targeted, sustained research effort. The program is developed around research Focus Areas: a topic area that is of high interest and priority to WRF subscribers because of a challenge or opportunity that is present, emerging, or anticipated, and for which research will help subscribers manage and address the challenge or optimize the opportunity. A focus area includes a discrete challenge or opportunity statement, measurable objectives, and one or more projects that will lead to applied solutions and benefits for WRF subscribers within a specified, relevant time frame. FOCUS AREA RFP This project is funded under the Focus Area titled, Waterborne Pathogens in Distribution and Plumbing Systems and is intended to support the Focus Area objective(s): To understand the factors governing the proliferation of waterborne pathogens. Method(s) for Detecting and Differentiating Opportunistic Premise Plumbing Pathogens (OPPPs) to Determine Efficacy of Control and Treatment Technologies (RFP #4721) Project Objective The objective of this project is to develop and/or optimize method(s) for detecting and differentiating multiple opportunistic premise plumbing pathogens and to demonstrate method applicability, suitability, and effective use following OPPP treatment and control options. Budget Proposals may request WRF funds in the range of $225,000 - $250,000. WRF funds requested and total project value will be a criterion considered in the proposal selection process. Background The presence of opportunistic premise plumbing pathogens (OPPPs) (e.g., Legionella pneumophila, nontuberculous mycobacteria, Acanthamoeba, Naegleria fowleri, Pseudomonas aeruginosa, to name a few) in drinking water distribution and plumbing systems has continued to grow in significance as a public health concern. However, the ability to provide reliable, sensitive and quantitative detection and differentiation of live and infectious pathogens remains a significant challenge for OPPPs. Currently, there is a lack of standardized methodologies or protocols for detecting and differentiating the variety of OPPPs potentially in potable water. The primary advantage of culture-based methods for detection of OPPPs remains their ability to confirm the presence of live pathogens. However, published data suggest that certain bacterial pathogens, such as Pseudomonas and Legionella, can under certain environmental conditions (e.g., presence of specific metal ions), survive in a viable but non-culturable (VBNC) state. There is also evidence that some strains of specific OPPPs present a larger risk to public health, especially to susceptible populations, but no methodologies to discriminate between pathogenicity currently exist. Free-living (trophozoic) protozoan organisms may play a critical role in biofilms as both primary pathogens and/or hosts for certain bacterial pathogens. They can also survive disinfection in a cystic state. Current detection methods provide limited ability to differentiate trophozoites from cysts, and even less ability to determine the infectivity of cysts.

2 Some molecular-based methodologies have been designed to distinguish non-viable cells from those in a dormant state. But these molecular methods have not been rigorously tested. While, molecular-based methods provide some distinct advantages including specificity and sensitivity, and the potential to produce high-throughput results, they are limited by the lack of both standardization and studies which directly compare their performance to that of other detection methods. Detailed information on OPPPs detection methodologies can be found in WRF report 4379 [1]. There is a need to develop reliable, sensitive, and quantitative method(s) for detecting and differentiating multiple opportunistic premise plumbing pathogens in order to demonstrate the effectiveness of OPPP treatment and control options. Several disinfection strategies are used for the control of a wide range of pathogens in drinking water distribution systems, including premise plumbing. Some studies have shown that the use of chloramine seems to reduce the frequency of L. pneumophila, but other studies have shown that this leads to an increase in the numbers of mycobacteria. Baron et al. (2014) observed a shift in the microbial ecology of a hospital s hot water system, following chloramination, using the Illumina 16S rrna sequencing population analysis [2]. However, the effectiveness of current control strategies is still not well documented because the methods for OPPPs have not been fully developed or validated. The deliverables of this project include the development of OPPP detection method(s) that are applicable and suitable tools for use in determining control and treatment effectiveness. In addition, the research team will evaluate the methods performance using at least one control option. Research Approach With a focus on known opportunistic pathogens in drinking water systems, select at least 3 OPPPs, and provide compelling reasons to support the selection. Complete a literature review to support the research direction. Propose the most appropriate fit-for-purpose method(s) for detecting and differentiating the opportunistic premise plumbing pathogens selected above. Demonstrate performance in a range of drinking water and premise plumbing (bulk water/biofilm) matrices. Because OPPPs can exist in several forms in nature (e.g., cyst, trophozoite, vegetative, viable but non-culturable, etc.), the proposed method(s) need to quantitatively differentiate between actively growing, dormant, and/or dead cells, etc. Appropriate positive controls and internal controls must be included. The developed method(s) should be evaluated for determining the effectiveness of OPPP treatments and control options. To do so, choose at least one control method/treatment/operation (e.g., chlorination, chloramination, heat treatment, copper-silver ionization UV, plumbing operation, etc.) to demonstrate the applicability of the detection method in documenting the effectiveness of the control option in either a bench or pilot test format. Prepare a report detailing the methodology, sensitivity, specificity, speed, accuracy, cost, and training required for the developed method(s). Consider holding a workshop or webinar to disseminate the methodology and provide training to water utility professionals.

3 Prepare a publishable report demonstrating the effectiveness of at least one control option, using the method(s) developed for OPPPs. References 1. A. Pruden, M. Edwards, and J. Falkinham (2013), State of the Science and Research Needs for Opportunistic Pathogens in Premise Plumbing, Water Research Foundation, Denver CO. 2. J. Baron, A. Vikram, S. Duda, J. Stout, and K. Bibby (2014), Shift in the Microbial Ecology of a Hospital Hot Water System following the Introduction of an On-Site Monochloramine Disinfection System, PLOS ONE, Vol. 9, Issue 7, e ~e Proposal Preparation Instructions Proposals submitted in response to this RFP must be prepared in accordance with the Water Research Foundation document Guidelines for Focus Area Program Proposals. The most current version of these guidelines is available at The guidelines contain instructions for the technical aspects, financial statements and administrative requirements that the applicant must follow when preparing a proposal. Eligibility to Submit Proposals This RFP solicits proposals from all technically qualified U.S. based or non-u.s. based applicants, including educational institutions, research organizations, federal or state agencies, local municipalities, and consultants or other for-profit entities. WRF s Board of Trustees has established a Timeliness Policy that addresses researcher adherence to project schedule. The policy can be reviewed at Researchers who are late on any ongoing WRF-sponsored studies without an approved no-cost extension are not eligible to be a named participant in any proposal. If you have any questions about your eligibility for WRF projects, please contact the WRF Research Manager listed at the bottom of the RFP. Administrative, Cost and Audit Standards WRF s standards for administrative, cost and audit compliance are based upon and comply with Office of Management and Budget (OMB) Uniform Grants Guidance (UGG), 2 CFR Part 200 Uniform Administrative Requirements, Cost Principles, and Audit Requirements for FederalAwards, and 48 CFR 31.2 Contracts with Commercial Organizations. These standards are referenced in the WRF s Guidelines for Focus Area Program Proposals and include specific guidelines outlining the requirements for Indirect Cost Negotiation Agreements, Financial Statements and the Statement of Direct Labor, Fringe Benefits and General Overhead. Inclusion of indirect costs must be substantiated by a negotiated agreement or appropriate Statement of Direct Labor, Fringe Benefits and General Overhead. Well in advance of preparing the proposal, your financial staff should review the detailed instructions included in WRF s annually released Guidelines for Focus Area Program Proposals.

4 Budget and Funding Information The funding available from WRF for this project is in the range of $225,000 - $250,000. A minimum 25 percent of the total project value must be contributed by the applicant (i.e. the applicant s minimum contribution must equal one-third of WRF funds requested). Acceptable forms of applicant contribution include cost-share, applicant in-kind or third-party in-kind that comply with 2 CFR Part Cost sharing or matching. The applicant may elect to contribute more than 25 percent to the project but the maximum WRF funding available remains fixed at $250,000. Proposals that do not meet the minimum 25 percent of the total project value will not be accepted. Period of Performance The proposed project schedule should be realistic, allowing ample time for the preparation of final reports and for review of project results. It is WRF s policy to negotiate a reasonable schedule for each research project. Once this schedule is established, WRF and its sub-recipients have a contractual obligation to adhere to the agreed-upon schedule. Under WRF s No-Cost Extension Policy, a project schedule cannot be extended more than nine months beyond the original contracted schedule, regardless of the number of extensions granted. The policy can be reviewed at Utility and Organization Participation WRF is especially interested in receiving proposals which include both participation and contribution of resources from water utilities and organizations in the research effort. Information on utilities and/or organizations that have indicated an interest in participating in this research project is attached. While WRF makes utility and organization participation volunteers known to applicants, it is the applicant s responsibility to negotiate utility and organization participation in their particular proposal, and the utilities and/or organizations are under no obligation to participate. Application Procedure and Deadline Proposals are now being accepted exclusively online in PDF only format and must be fully submitted before June 14, 2017, 5pm Mountain Time. All the forms and components of the proposal are available online in the Proposal Component Packet zip file. A login is required to download this packet and use the proposal website. This information is available at The online proposal system allows submission of your documents until the date and time stated in the RFP. To avoid the risk of the system closing before you press the submit button, do not wait until the last minute to complete your submission. Questions to clarify the intent of this Request for Proposals and WRF s administrative, cost and financial requirements may be addressed to the Research Manager, Grace Jang, at (303) or by at hjang@waterrf.org.

5 UTILITY AND ORGANIZATION PARTICIPANTS The following utilities have indicated an interest in possible participation in this research. This information is updated within 24 business hours when a utility submits a volunteer form and this RFP will be re-posted with the new information. (Depending upon your settings, you may need to click refresh on your browser to load the latest file.) Sarah Wright Environmental Laboratories Senior Specialist Association of Public Health Laboratories 8515 Georgia Avenue Suite 700 Silver Spring, MD (240) sarah.wright@aphl.org Aspa Capetanakis Technical Assistant NYC DEP Horace Harding Expressway Corons, NY (718) (718) acapetanakis@dep.nyc.gov Jaime Fleming Laboratory Manager City of Wyoming Water Treatment Plant New Holland St Holland, MI (616) flemingj@wyomingmi.gov