CWRS Advisory Board Meeting March 1, 2007 Gretchen Grover – Chemist Christal Wade - Microbiologist Rick Fowler – Environmental Biotechnologist.

CWRS Advisory Board MeetingCWRS Advisory Board MeetingMarch 1, 2007March 1, 2007

Gretchen Grover – ChemistGretchen Grover – ChemistChristal Wade - MicrobiologistChristal Wade - Microbiologist

Rick Fowler – Environmental BiotechnologistRick Fowler – Environmental Biotechnologist

WATERS EmployeesWATERS Employees Director: Director: Jana FatticJana Fattic Lab Analysts:Lab Analysts:

Microbiology Department: Rick Fowler, Christal Microbiology Department: Rick Fowler, Christal WadeWade

Chemistry Department: Gretchen Grover, Chemistry Department: Gretchen Grover, Dong LiDong Li

Office Associate: Office Associate: Kristy MayesKristy Mayes Students: Students: Samantha Kramer, Rebekkah Lively, Samantha Kramer, Rebekkah Lively,

Jennifer DeanJennifer Dean

CustomersCustomers Private

Total Coliform/E. Coli, Metals, Ions, AtrazineTotal Coliform/E. Coli, Metals, Ions, Atrazine Wastewater

BOD, pH, TRC, DO, TSS, Fecal Coliform, Ammonia BOD, pH, TRC, DO, TSS, Fecal Coliform, Ammonia NitrogenNitrogen

Storm water COD, Oil and Grease, TDS, TSSCOD, Oil and Grease, TDS, TSS

Drinking water Total Coliform/E.ColiTotal Coliform/E.Coli

Oil Wells pH, TDS, Specific Gravity, Chloride pH, TDS, Specific Gravity, Chloride

Research Projects TOC, Atrazine, Metals, Ions, DNATOC, Atrazine, Metals, Ions, DNA

CertificationsCertifications Drinking water CertificationDrinking water Certification

January AuditJanuary Audit Inorganic CertificationInorganic Certification

June AuditJune Audit Cryptosporidium Cryptosporidium

Long Term 2 Enhanced Long Term 2 Enhanced Surface Water Treatment Surface Water Treatment Rule (LT2)Rule (LT2)

Accept outside samples under Accept outside samples under LT2 and provides field LT2 and provides field filtration apparatusfiltration apparatus

www.epa.gov/safewater/disinfection/www.epa.gov/safewater/disinfection/LT2/lab_aprvlabs.htmlLT2/lab_aprvlabs.html

Chemical Hygiene PlanChemical Hygiene Plan Chemical Hygiene OfficerChemical Hygiene Officer

ProjectsProjects Mammoth CaveMammoth Cave

Flood Pulse EventFlood Pulse Event City of Bowling GreenCity of Bowling Green

River & Cave System monitoringRiver & Cave System monitoring Watershed WatchWatershed Watch

Upper Green River watershed monitoring (E.coli)Upper Green River watershed monitoring (E.coli) TMDLTMDL

Analyzing samples for this projectAnalyzing samples for this project University ResearchUniversity Research

Support WKU faculty as well as other university Support WKU faculty as well as other university projectsprojects

Quality AssuranceQuality Assurance Continued revision and updating of Continued revision and updating of

SOP to the 21SOP to the 21stst Edition Standard Edition Standard Methods Methods

QA/QC ImprovementsQA/QC Improvements New StandardsNew Standards

PT/QC (quarterly)PT/QC (quarterly) USGSUSGS APGAPG

Important Important PurchasesPurchases

ChemistryChemistry Accumet excel XL60Accumet excel XL60

TKN DigestorTKN Digestor KonelabKonelab

MicrobiologyMicrobiology Bench top water bath

FastGel electrophoresis apparatus PCR Workstation

Qubit fluorometer Microcentrifuge

Where do we go from here?Where do we go from here? Current CapabilitiesCurrent Capabilities

CryptosporidiumCryptosporidium Konelab AnalysesKonelab Analyses Drinking water, Wastewater, Storm water analysesDrinking water, Wastewater, Storm water analyses Bacteriological/DNA analysesBacteriological/DNA analyses

Future CapabilitiesFuture Capabilities Unregulated Contaminant Monitoring Rule (UCMR2)Unregulated Contaminant Monitoring Rule (UCMR2) Organics (Work toward a GC MS/MS and HPLC)Organics (Work toward a GC MS/MS and HPLC) RadionuclidesRadionuclides Direct a Chemical Laboratory Certification Program Direct a Chemical Laboratory Certification Program

for Kentuckyfor Kentucky Voluntary Wastewater CertificationVoluntary Wastewater Certification

Where do we go from here?Where do we go from here?

New customers bring more cash flowNew customers bring more cash flow 11 new customers 11 new customers Networking; customer surveysNetworking; customer surveys Pick up servicePick up service Consultation servicesConsultation services Sample collection servicesSample collection services

Microbiology Microbiology ProgramsPrograms

Christal WadeChristal Wade

Drinking Water MicrobiologyDrinking Water Microbiology

Sample Type Analyzed Number Analyzed/year

Drinking Water (Public) 2080

Drinking Water (Private) 25

Wastewater 0

Surface (raw) 745

Total 2850

Laboratory Sample Load

Completed latest On-Site Audit in January 2007

EPA Approval for CryptoEPA Approval for Crypto WATERS Lab received approval from WATERS Lab received approval from

EPA for the analysis of Cryptosporidium EPA for the analysis of Cryptosporidium on October 18, 2006.on October 18, 2006.

Currently have a cooperative Currently have a cooperative agreement with Appalachian States agreement with Appalachian States Analytical, L.L.C.Analytical, L.L.C. Total of 20 surface water clientsTotal of 20 surface water clients Will be starting monitoring in JuneWill be starting monitoring in June

4 Contracts with other PWSs4 Contracts with other PWSs From both Kentucky and West VirginiaFrom both Kentucky and West Virginia Several quotes outstandingSeveral quotes outstanding

Latest PT ResultsLatest PT ResultsFrom September 2006From September 2006

Oocysts Reported

Oocysts Spiked

% RecoveredCysts

ReportedCysts

Spiked%

Recovered

03604 152 195 78 100 201 50

03605 139 195 71 91 201 45

03606 138 180 77 0 0 N/A

75 48

5 7RSD of % Recovery

EPA Protozoan Proficiency Testing Laboratory Results Summary

Lab ID: KY00002Round: 0906 Filter: Envirochek HV

Sample ID

Cryptosporidium Giardia

Mean % Recovery

Completed another set of PT samples in February 2007

Mean (%) Median (%) RSD (%) Mean (%) Median (%) RSD (%)

Envirochek 1 35 35 31 41 41 21

Envirochek HV 45 45 43 42 46 48 27

Filta-Max 15 47 46 35 39 43 44

Filta-Max Xpress 4 29 27 34 29 26 41

Direct Centrifugation

1 61 61 1 49 49 25

Continuous Flow Centrifugation

1 69 68 5 62 62 5

CrypTest 1 59 65 23 5 5 141

All Versions 68 45 44 41 43 46 35

WATERS Lab 75 5 48 7

OPT Recovery Statistics for Each Method Variation

Giardia RecoveriesCryptosprodium RecoveriesMethod Version # Labs

Cryptosporidium OPR

0.0

0.2

0.4

0.6

0.8

1.006

247-

02

0625

1-02

0626

4-02

0627

8-02

0629

2-02

0630

6-02

0632

0-02

0633

4-02

0634

8-02

0636

2-02

0701

1-02

0702

5-02

0703

8-02

Sample Identification

Per

cen

t R

eco

very

Ongoing Precision and Recovery

Mean Recovery 52.4%

Giardia OPR

0.00.10.20.30.40.50.60.70.80.91.0

06247-0

2

06251-0

2

06264-0

2

06278-0

2

06292-0

2

06306-0

2

06320-0

2

06334-0

2

06348-0

2

06362-0

2

07011-0

2

07025-0

2

07038-0

2

Sample Identification

Per

cen

t R

eco

very

Ongoing Precision and Recovery

Mean Recovery 41.1%

Education & OutreachEducation & Outreach 2 Hour Crypto Presentation2 Hour Crypto Presentation

Approved for Drinking Water Operator Approved for Drinking Water Operator CEUCEU

Presented at KRWA Training SessionsPresented at KRWA Training Sessions Will Present at KWWOA Annual Will Present at KWWOA Annual

ConferenceConference 1 Hour 1 Hour E. coliE. coli Presentation Presentation

Approved for Waste Water Operator CEUApproved for Waste Water Operator CEU Presented at KRWA Training SessionsPresented at KRWA Training Sessions Will Present for KWWOAWill Present for KWWOA

Environmental Environmental BiotechnologyBiotechnology

RickRick FowlerFowler

DNA Analysis ServicesDNA Analysis Services DNA extraction from water, soil, membrane DNA extraction from water, soil, membrane

filters, pure cultures, swabs, etc.filters, pure cultures, swabs, etc. Yield of total DNA measured with Qubit Yield of total DNA measured with Qubit

fluorometer, sensitivity 0.2 x 10fluorometer, sensitivity 0.2 x 10-12 -12 gg Quantitative Real-Time PCRQuantitative Real-Time PCR

Determines concentration of DNA from specific Determines concentration of DNA from specific organisms in a mixture, sensitivity <1 pptorganisms in a mixture, sensitivity <1 ppt

DNA cloning and sequencingDNA cloning and sequencing Cloning permits segregation of individual Cloning permits segregation of individual

species comprising environmental microbial species comprising environmental microbial communities communities

DNA sequencing yields unique gene sequences DNA sequencing yields unique gene sequences for identification of organismsfor identification of organisms

Bacteria

Parasites Algae

Fungi Amoebae

Zooplankton

Limestone

Ca13CO3

Field work in Mammoth Field work in Mammoth CaveCave

Examples of clients Examples of clients supported by WATERS DNA supported by WATERS DNA

servicesservices Louisville Water CompanyLouisville Water Company Ammonia Oxidizing Bacteria in distribution systemAmmonia Oxidizing Bacteria in distribution system AcanthamoebaAcanthamoeba in eyewash stations in eyewash stations

KSU Aquaculture Research CenterKSU Aquaculture Research Center DNA analysis of bacteria, algae, and fungi used to DNA analysis of bacteria, algae, and fungi used to

evaluate conditions for optimal shrimp productionevaluate conditions for optimal shrimp production Fungi in some tanks identified by cloning and DNA Fungi in some tanks identified by cloning and DNA

sequencing to assess potential risk or benefitsequencing to assess potential risk or benefit National Cave and Karst Research InstituteNational Cave and Karst Research Institute

Develop a universal monitoring scheme applicable to Develop a universal monitoring scheme applicable to caves nationwide and catalog any new caves nationwide and catalog any new microorganisms indigenous to cave climatesmicroorganisms indigenous to cave climates

Examples, continuedExamples, continued Mammoth Cave National ParkMammoth Cave National Park

Ongoing monitoring of bacterial communities for Ongoing monitoring of bacterial communities for better understanding of karst aquifers and as better understanding of karst aquifers and as indicators of environmental changesindicators of environmental changes

Monitor Monitor Chlorella Chlorella DNA in surface waters to assist DNA in surface waters to assist with protection of endangered mussellswith protection of endangered mussells

Oregon Caves National MonumentOregon Caves National Monument Cloning and sequencing of bacterial and fungal Cloning and sequencing of bacterial and fungal

communities on- and off-tourist trails to assess communities on- and off-tourist trails to assess environmental impacts of tourismenvironmental impacts of tourism

Abundance of iron-sulfur bacteria indicate growth Abundance of iron-sulfur bacteria indicate growth in absence of light is derived from chemical energy in absence of light is derived from chemical energy

Scientific Outreach, Scientific Outreach, Presentations, and Presentations, and

WorkshopsWorkshops National Speleological SocietyNational Speleological Society

2001, 20062001, 2006 National Cave and Karst Research National Cave and Karst Research

InstituteInstitute 2001, 20052001, 2005

Geological Society of AmericaGeological Society of America 2001, 2004 2001, 2004

National Park Service advisory panelsNational Park Service advisory panels 2001, 2002, 2003, 2004, 2005, 20062001, 2002, 2003, 2004, 2005, 2006

ISEISISEIS 20062006