Dreissena Mussel Early Detection Monitoring Methods and Quality Assurance Workshops:

1. QZAP 2010 Project Updates CRB Working Group 6 June 2012 Dreissena Mussel Early Detection Monitoring Methods and Quality Assurance Workshops: Two Workshops Addressing Critical Impediments to the Expansion of Western Early Detection Monitoring Programs Pilot Laboratory Testing Program For the Early Detection of Zebra and Quagga Mussels in Western U.S. Waters Kevin L Kelly – US Bureau of Reclamation Sandra A. Nierzwicki-Bauer – Darrin Fresh Water Institute, Rensselaer Marc E Frischer – Skidaway Institute of Oceanography

2. 2012 Dreissena Mussel Early Detection Monitoring Methods & Quality Assurance WorkshopsFort Worth, Texas 7-10 February 2012 Texas Christian University Workshops attracted 41 participants from 25 different Agencies and Institutions. Executive Summary available at: http://www.musselmonitoring.com/2012Workshop/ExecSummary.pdf Ranked research and action priorities http://www.musselmonitoring.com/2012Workshop/Ranked_RRII_recommendations_%28FINAL%29.pdf The final workshop report is still in preparation

3. Dreissena Early Detection Best Practices Technical Workshop Primary Goal (Deliverable) Develop Recommendations for Best Practice Protocols For Dreissena spp. Presence/Absence Detection & Quantification By CPLM, IFC & PCR SOPs can be written for each method. Will be based on equivalency to defined detection limits – 10-50 m-3 (0.01-0.05 l-1) Key priorities for improving early detection methods and procedures were identified

4. Dreissena Early Detection Laboratory Standards Technical Workshop Primary Goal (Deliverable) Establish a consensus regarding the need for a laboratory accreditation program and the scope of such a program. Develop a roadmap for the development and implementation of such a program

5. One recommendation was to develop a taxonomy certification program to help identify personnel that are qualified to identify Dreissena spp. larvae in plankton tow samples microscopically Partner with the Society For Freshwater Science (SFS) To develop a certification exam for Dreissena spp. Process is currently stalled. We need to identify a group and a mechanism to develop a consensus among our microscopy experts that will develop a list of taxa and life history stages for a certification exam This group, working with the SFS taxonomy certification committee will develop and administer Dreissena spp. identification certification. Can we make this happen?

6. Pilot Laboratory Testing Program For the Early Detection of Zebra and Quagga Mussels in Western U.S. Waters Three Project Components: Sample Preservation Study: What is the best way to store plankton tow samples for eventual examination for Dreissena spp. larvae using microscopic and PCR-based approaches? Optimal sampling strategies? To detect rare veligers is it better to collect multiple samples at a single high risk location or to randomly sample over a larger geographic area? Develop a pilot scale laboratory testing program that would allow laboratories to obtain blind samples that could be used for internal quality control, assay development and competence training.

7. Sample Preservation Study 1 1 1 Unpreserved samples initially frozen

10. pH ~ 7.8 pH ~ 9.5

11. Preliminary Evaluation (Microscopy) No fixation results in significant sample decay (not surprising) All the concentrations of EtOH used seem to be effective (a bit surprising) pH adjustment doesn't necessarily hurt but makes sample examination difficult and in these samples at least appears to be unnecessary Freezing at either -20 or -60/-80 results in sample decay (surprising)

12. Transect #2 Fish Hatchery – Transect #1 Marina – Transect #3 Transect #4 Sampling Strategy Question arose at 2009 workshops: What is the best sampling strategy for detecting rare veligers? Multiple locations of plankton tows vs. multiple plankton tows from a single (high risk) location?

13. Preliminary Conclusions At least in a fast flowing river environment there does not seem to be a difference in the detection of rare veligers whether samples are collected from a single location (in this case a transect across the river at a single location) versus collecting the same number of samples along a river section (in this case a 10 km river segment) Caveats: Larvae are likely to be more evenly distributed in a fast flowing river than in a lake (However, we’ve observed patchy larval distribution in Hudson River) Larval concentrations in Colorado River at Willow Beach site were quite high when we sampled. Would like to repeat these studies in lakes and at lower larval concentrations. we might get a chance to do this later this summer in NY.

14. Laboratory Testing Program For Early Detection of Zebra & Quagga Mussels Program Components: Provide “certified” larval samples to: • Help advance method development research • Provide “standards” for labs to independently assess their results • Provide “blind samples” to labs to help identify weaknesses in existing methods

15. Reference Materials Distribution of Samples • Distribution of “certified” dreissenid larval samples to: • 1. Analytical laboratories 2. Agencies contracting analytical services 3. Monitoring programs • Request form will be available online • Independently validated “reference” samples containing: • D. polymorpha, D. bugensis (or both) with known number of veligers spiked into plankton matrix (provided by requestor or supplier) • Blind spiked samples: • Spiked with larvae over a range of concentrations (0-25 larvae/25 ml) • Prepared in either plankton matrix provided by requestor or supplier (Dreissena-free prior to spiking)

16. Laboratory Testing Program – Status & Timeline • Collection of materials to be used for preparation of Reference materials • Quagga Veligers (February 2011) • Zebra Veligers from Lake Champlain, Glen Lake (August 2011) • Plankton material (with no zebra or quagga veligers) (November 2011) • All materials are archived at Darrin Fresh Water Institute • Materials are stored at 4ºC • Draft of “request of materials” web form. Final version June 15, 2012 • Request of materials on-line June 30, 2012 • Begin sample distribution upon request July ,2012

17. Action Items • Final project reports completed by Sept 2012 or earlier • Need to act on workshop recommendations • Method SOPs need to be written for CPLM, IFC and PCR • Efforts to continue methods research (methods cook off) • Pilot laboratory testing program implemented and run for at • least one year (no additional cost to this round of QZAP funding) • Develop microscopy certification program with SFS