DNADEQ

DNA Fingerprinting of Bacteria Sources in the Tualatin Sub-basin: 

DNA Fingerprinting of Bacteria Sources in the Tualatin Sub-basin Jan Miller Clean Water Services 2005

Background: 

Background 2001 Tualatin Sub-basin Total Maximum Daily Load (TMDL) established Wasteload Allocations (WLA) for bacteria in stormwater Clean Water Services needs to develop Best Management Practices (BMPs) to reduce bacteria in stormwater to meet its Wasteload Allocations

Bacteria TMDL: 

Bacteria TMDL Addresses Public Health Issues Water Contact Recreation E. coli is the indicator organism It is not a pathogen Indicates possible presence of pathogens Bacteria such as Salmonella and Shigella Protozoa such as Giaradia or Cryptosporidia Viruses

Wasteload Allocations: 

Wasteload Allocations Summer Levels for Stormwater 3,000 to 12,000 E. coli per 100 mLs depending on the subbasin Winter Levels for Stormwater 700 to 11,000 E. coli per 100 mLs depending on the subbasin All other times in summer and winter 406 E. coli per 100 mLs

Bacteria Loads Compared to WLASpatially Averaged by PLOAD: 

Bacteria Loads Compared to WLA Spatially Averaged by PLOAD

E.coli levels in Stormwater for 2000 to 2005: 

E.coli levels in Stormwater for 2000 to 2005 Wasteload Allocations

E. coli values from 2000 and 2005Stream Study Sites: 

E. coli values from 2000 and 2005 Stream Study Sites

Bacteria BMPs: 

Bacteria BMPs Connect septic systems to sanitary systems Actively manage pet waste Minimize the impact of hobby farms Limit duck and goose waste Clean up illegal dump sites

Need Change in Public Behavior: 

Need Change in Public Behavior Many of the BMPs require behavior changes by the public Public needs proof their activities are causing problems Thanks to court cases and CSI-style programs - DNA is accepted as “proof” by the public

Microbial Source Tracking (MST): 

Microbial Source Tracking (MST) Institute for Environmental Health (IEH) Formerly University of Washington Ribosomal RNA typing method Cost for typing DEQ 319h Grant $41,723 in 2002 Clean Water Services $40,000 Cost for sampling and E. coli isolation Clean Water Services $11,000 Urban area of Tualatin Sub-basin

Sampling Methodology: 

Sampling Methodology Sites 5 Stream Sites representing areas of potential concern 3 Stormwater Sites 2 Duplicates (one stream and one stormwater) Weather Conditions Summer Dry period Summer Storm Winter Dry Period Winter Storm

Goal 100 Isolates for Statistical Reliability: 

Goal 100 Isolates for Statistical Reliability 6 Stormwater or 7 stream events sampled 5 Samples collected per event 3 E. coli isolates tested per sample 90 to 105 isolates per site 6 x 5 x 3 = 90 7 x 5 x 3 = 105

Sites Sampled: 

Sites Sampled Stream Sites Residential Area – Septic Tank Area Residential Area – High Dog Population Residential Area – Older Urban Down stream of a pond – Duck Feeding Residential area – Mixed Urban Stormwater Commercial Parking Lot New Residential Area Water Quality Facility Effluent (Vegetated Pond)

Distribution and Grouping of Sources: 

Distribution and Grouping of Sources Source ID Example Using Canine: Dog means dog Coyote means coyote Canine means the DNA type is shared between dog, coyote, wolf, fox etc. Additional Sources Feline 13 Unknown 85 Total Isolates 924

All Sites All Events(No Duplicates): 

All Sites All Events (No Duplicates) 924 Isolates

Individual Sites: 

Individual Sites

Stream - Septic Tank Area: 

Stream - Septic Tank Area 134 Isolates

Stream - High Dog Population: 

Stream - High Dog Population 125 Isolates

Stream - Older Urban: 

Stream - Older Urban 131 Isolates

Stream - Duck Feeding : 

Stream - Duck Feeding 133 Isolates

Stream - Mixed Urban: 

Stream - Mixed Urban 130 Isolates

Stream – Duck FeedingDuplicates: 

Stream – Duck Feeding Duplicates Sample Duplicate

Stormwater – Commercial Parking Lot: 

Stormwater – Commercial Parking Lot 79 Isolates

Stormwater – New Residential Area : 

Stormwater – New Residential Area 96 Isolates

Stormwater – Water Quality Facility Effluent : 

Stormwater – Water Quality Facility Effluent 96 Isolates

Stormwater – New Residential Area Duplicates: 

Stormwater – New Residential Area Duplicates Sample Duplicate

Compare Percentages for each Site with by Key Sources: 

Compare Percentages for each Site with by Key Sources

No Statistical Difference Between Sites for Source: 

No Statistical Difference Between Sites for Source Statistical test for outliers Calculate the difference between a data point and the nearest value as a fraction of the range from the largest to the smallest value in the data set Compare result with a statistical table 5% chance of incorrect conclusion

Evaluate Stream Samples Only: 

Evaluate Stream Samples Only

Stream Bacteria Levels During Study: 

Stream Bacteria Levels During Study

Stream Bacteria Sources – All Sites: 

Stream Bacteria Sources – All Sites 653 Isolates

Stream Bacteria Primary SourcesPercentage By Weather Condition: 

Stream Bacteria Primary Sources Percentage By Weather Condition

Evaluate Stormwater Samples Only : 

Evaluate Stormwater Samples Only

Stormwater Runoff Bacteria Levels During Study: 

Stormwater Runoff Bacteria Levels During Study

Stormwater Bacteria Sources – All Sites: 

Stormwater Bacteria Sources – All Sites 271 Isolates

Stormwater Bacteria from Primary SourcesPercentage by Weather Condition: 

Stormwater Bacteria from Primary Sources Percentage by Weather Condition

Compare Results with Other Studies: 

Compare Results with Other Studies

Comparison to Other StudiesPercentage by Type: 

Comparison to Other Studies Percentage by Type

What Does It All Mean: 

What Does It All Mean Diverse Set of Sites Similar Distribution of Sources Different Event Types Similar Distribution of Sources Different Studies Similar Distribution of Sources

Relationship to Wasteload Allocations: 

Relationship to Wasteload Allocations Provides evidence that humans are not a dominant source of bacteria in streams This is a policy issue Provides evidence that human activities are resulting in increased bacteria in streams No obvious solution indicated Many individual actions Too much variability to predict percent reduction with combinations of actions

Public Education: 

Public Education Some things will be a difficult “sell” The biggest problem – Birds May get some support for signs that ask people not to feed the ducks and geese May get some support for changes in landscaping to discourage ducks and geese But The public likes ducks and geese They like other types of birds even more!

Public Education Creativity: 

Public Education Creativity First Effort using this study City of Tigard Carla Staedter

Holy Dog Poop Batman!!!! : 

Holy Dog Poop Batman!!!! How much dog poop is “natural” Given that the wolf is the ancestor of dogs How many wolves can the Tualatin Sub-basin support? 70 wolves There are 80,000 dogs in the Tualatin Sub-basin That leaves 79,930 dogs to pick up after!

Prototype of Sign : 

Prototype of Sign

What’s Next: 

What’s Next Reality check on what the Bacteria TMDLs are trying to accomplish Reality check on where we should focus our efforts Human sources YES Human activities Provide the public with knowledge and tools to help the environment

The End: 

The End