microbial source tracking - great lakes 2012

7/30/2019 Microbial Source Tracking - Great Lakes 2012

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Microbial Source Tracking:In Search of Human, Gull, and

Dog

Mauricio Larenas

12th Annual Great Lakes BeachAssociation Conference

October 16-18, 2012


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Fecal Indicators

Used to determine the presence of pathogenic organisms

Typical Fecal IndicatorsE. ColiEnteroccocus

Fecal ColiformBacteroidales (EPA Method B)

Useful for the detection of general fecal pollution

Provide no insight into the potential sources of fecal

pollution

Microbial Source TrackingIn Search ofHuman, Gull, and Dog

Fecal pollution problem

Microbial sourcetracking plan

Submitting and testingsamples

Project results andoutcomes

Conclusion


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Pathways for Contamination

Industrial PlantsFarm wasteUrban runoff SewageSeptic LeaksWildlife




Submitting and testingsamples


Conclusion


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ProblemState of Delaware wasobserving high levels of Enterococcus at an

estuarine inlandrecreational beach




Submitting and testing

samples


Conclusion

Project Case Study


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Project Case Study

GoalDetermine if Human , Dog or Seagull fecalsources contributed to the high Enterococcuslevels





samples


Conclusion


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FundingFunding source for Delaware project:

EPA Region 3 BEACH Act grantDelaware Environmental Laboratory

Michigan funding:Nonpoint Source Program Grants

DESCRIPTION: Michigan's Department of Environmental Qualityadministers a Nonpoint Source (NPS) Program that assists state,federal, and local partners to restore water bodies impaired byNPS pollution and protect high quality waters from impairmentscaused by NPS pollution.

Michigan Sea GrantDESCRIPTION: Michigan Sea Grant, a cooperative program of theUniversity of Michigan and Michigan State University, is part of theNational Sea Grant College Program. It aims to promote betterunderstanding, conservation and use of Michigans coastalresources. Michigan Sea Grant funds integrated research projectsdesigned to foster science-based decisions about the use andconservation of Great Lakes resources.





samples


Conclusion


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Validate the sensitivity of two biomarkers

Human Polyomavirus- USFRaw untreated wastewater from 3 wastewater treatmentfacilities in Sussex County, DE

ResultsSewage tested positive for HPyVs

Gull Biomarker - USF/EPA37 fecal samples from herring and/or laughing gulls testedindividually

Results84% of the samples tested positive for the gull associatedmarker

Validation of local fecal samplesMicrobial Source TrackingIn Search ofHuman, Gull, and Dog




samples


Conclusion


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PlanUse 1 Seagull, 1 Dog, and 2 Human tests todetermine which host(s) was contributing tofecal contamination49 Samples taken through the summer monthsand sent to Source Molecular for analysis

Equal number of samples analyzed by USFSamples taken in knee deep water 3 times a week at

3 locations





samples


Conclusion

Project Case Study


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PlanDNREC would test for general fecal indicator

Enterococcus concentrations - Enterolert





samples


Conclusion

Project Case Study


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Submitting SamplesMicrobial Source TrackingIn Search ofHuman, Gull, and Dog




samples


Conclusion

Samples where filtered at the Delaware DNREClaboratory and submitted frozen on dry ice

Samples typically submitted as 500mL water samplesovernight

Sterile bottles and packing material provided in mostcases

Serves as convenience and to maintain QA/QC


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Microbial Source Tracking

Host gut conditions and natural selection create uniquegenetics between host microbial communities.

Cow and Chicken microbial communities are not the same

Genetic differences are identified and used as the basis forfecal host discrimination

Host specific genes are detected using qPCRExponentially copies the sequence of interest.Monitors and calculates the starting quantity





samples


Conclusion


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What hosts can MST detect?Microbial Source TrackingIn Search ofHuman, Gull, and Dog




samples


Conclusion


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Presence/Absence MST ResultsMicrobial Source TrackingIn Search ofHuman, Gull, and Dog




samples


Conclusion

Turnaround time: 5-10 working daysDetermines +/- of host specific fecal indicators

Multiple tests targeting the same host (e.g. Human) allow confirmationof results

Valuable for screening multiple hosts prior to quantification

Less expensive than quantification


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Quantification MST Results

Turnaround time: 5-10 working days

Reports the genetic copiesfrom host specific fecalindicators

These values are used tointerpret the hosts relativecontribution to fecalcontamination

Results reported as:NegativeTrace

Minor ContributorMajor Contributor

A general fecal indicator canalso be quantified todetermine the generalcontamination





samples


Conclusion


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ResultsDog

4% of the samples were positiveQuantification values showed few dog specific genetic

copiesHuman Bacteroidetes (HF183)

8% of the samples were positiveQuantification values showed few human specificgenetic copies

Seagull98% of the samples were positive

Quantification values showed abundant seagull specificgenetic copies





samples


Conclusion

Project Case Study


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Project conclusions and outcomesSeagull was likely a major contributor the fecalpollution detected at the beach

Human was a possible contributor but occurringsporadically

Dog was not a likely fecal pollution contributor

The purpose of the MST study was to confirm the

visual observations that seagulls contributed to theelevated levels of Enterococcus (Delaware s waterquality indicator) at this beach. Based on MST results,there will be no change to management actions atthe site.





samples


Conclusion

Project Case Study


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Project Considerations

Are markers available for the potential fecal sources in thewater system?

Are the markers properly validated?Cross reactivity and sensitivity for those markers

What is the scale and budget for the project?

Microbes used for MST markers may not have a directcorrelation to traditional fecal indicators





samples


Conclusion


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Microbial Source TrackingMicrobial Source TrackingIn Search ofHuman, Gull, and Dog




samples


Conclusion


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About Source Molecular

Established in 2002 to fill the void between research andimplementation

Active in the research and development of newapproaches

Strict focus on Microbial Source Tracking

Detection of human, cattle, swine, bird, gull, goose, chicken,dog, deer, ruminant and horse possible

Pending ISO 17025 accreditation





samples


Conclusion


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4985 SW 74th CourtMiami, Florida 33155 USATelephone: (786) 220-0379

Mauricio [email protected]

Case Study: Microbial Source Tracking at Tower Road Bayside Beach, Rehoboth Bay, Sussex County, Delaware2011 (Dr. Humphries, DE DNREC; Dr. Harwood, USF; and Mr. Larenas, Source Molecular)

microbial source tracking - great lakes 2012

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