Dreissena Mussel Early Detection Monitoring Methods andQuality Assurance Workshops:

Two Workshops Addressing Critical Impediments to the Expansion ofWestern Early Detection Monitoring Programs

Pilot Laboratory Testing ProgramFor the Early Detection of Zebra and Quagga Mussels

in Western U.S. Waters

QZAP 2010 Project UpdatesCRB Working Group

6 June 2012

Kevin L Kelly – US Bureau of Reclamation

Sandra A. Nierzwicki-Bauer – Darrin Fresh Water Institute, Rensselaer

Marc E Frischer – Skidaway Institute of Oceanography

2012 Dreissena Mussel Early Detection Monitoring Methods & Quality Assurance Workshops

Fort Worth, Texas 7-10 February 2012

Texas Christian University

http://www.musselmonitoring.com/2012Workshop/Ranked_RRII_recommendations_%28FINAL%29.pdf

http://www.musselmonitoring.com/2012Workshop/ExecSummary.pdf

Workshops attracted 41 participants from 25 differentAgencies and Institutions.

Executive Summary available at:

Ranked research and action priorities

The final workshop report is still in preparation

Dreissena Early Detection Best Practices Technical Workshop

Primary Goal (Deliverable)Develop Recommendations for Best PracticeProtocols For Dreissena spp. Presence/AbsenceDetection & Quantification By CPLM, IFC & PCR

SOPs can be written for each method. Will be basedon equivalency to defined detection limits – 10-50 m-3 (0.01-0.05 l-1)

Key priorities for improving early detection methodsand procedures were identified

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 andimplementation of such a program

One recommendation was to develop a taxonomy certificationprogram to help identify personnel that are qualified to identifyDreissena 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 identifya group and a mechanism to develop a consensusamong our microscopy experts that will develop a listof taxa and life history stages for a certification exam

This group, working with the SFS taxonomy certificationcommittee will develop and administer Dreissena spp.identification certification.

Can we make this happen?

Sample Preservation Study: What is the best way to store planktontow samples for eventual examination for Dreissena spp. larvae usingmicroscopic and PCR-based approaches?

Pilot Laboratory Testing ProgramFor the Early Detection of Zebra and Quagga Mussels

in Western U.S. Waters

Three Project Components:

Optimal sampling strategies? To detect rare veligers is it better tocollect multiple samples at a single high risk location or to randomlysample over a larger geographic area?

Develop a pilot scale laboratory testing program that would allowlaboratories to obtain blind samples that could be used for internalquality control, assay development and competence training.

Sample Preservation Study

Unpreserved samples initially frozen

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pH ~ 9.5pH ~ 7.8

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 examinationdifficult and in these samples at least appears to be unnecessary

Freezing at either -20 or -60/-80 results in sample decay (surprising)

Sampling Strategy

Question arose at 2009 workshops: What is the best sampling strategy fordetecting rare veligers?

Multiple locations of plankton tows vs. multipleplankton tows from a single (high risk) location?

Fish Hatchery – Transect #1

Marina – Transect #3

Transect #2

Transect #4

At least in a fast flowing river environment there does not seem to be adifference in the detection of rare veligers whether samples are collectedfrom 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 riverthan in a lake (However, we’ve observed patchy larval distribution inHudson River)

Larval concentrations in Colorado River at Willow Beach site werequite 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.

Preliminary Conclusions

Laboratory Testing Program For Early Detection of Zebra & Quagga Mussels

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

Program Components:

Reference Materials

1. 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)

2. 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)

Distribution of Samples

Distribution of “certified” dreissenid larval samples to:1. Analytical laboratories2. Agencies contracting analytical services3. Monitoring programs• Request form will be available online

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

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