Pathogenicity Potential in Selected Locations of Rapid Creek Dr. - - PowerPoint PPT Presentation

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Pathogenicity Potential in Selected Locations of Rapid Creek Dr. - - PowerPoint PPT Presentation

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Pathogenicity Potential in Selected Locations of Rapid Creek Dr. Kelsey E. Murray Dr. Lisa A. Kunza Dr. Linda C. DeVeaux Curr rrent EP EPA A Water Quality ty T Testing Metric f for r Fecal Conta tamination of R Recreational Water

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Pathogenicity Potential in Selected Locations

  • f Rapid Creek
  • Dr. Kelsey E. Murray
  • Dr. Lisa A. Kunza
  • Dr. Linda C. DeVeaux
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SLIDE 2

Curr rrent EP EPA A Water Quality ty T Testing Metric f for r Fecal Conta tamination of R Recreational Water

  • Total fecal

coliforms

  • E. coli
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SLIDE 3

Escherichia c coli: A Geneti tically Diverse Species of B Bacteri ria

3

Environmental and naturalized Commensal to mammalian intestinal tracts; Probiotics Highly adapted strains that allow for a broad spectrum of human disease

  • Routine testing provides

a snapshot microbial abundance and content, but not pathogenic profile of the bacteria

  • The difference lies

within the genes (DNA)

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SLIDE 4

The Promiscuity ty of B Bacteri ria

4

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Healt lth Ris isks A Associa iated wit ith E. c coli

  • STEC = Shiga-toxigenic E. coli
  • Shiga toxins: cytotoxins that inhibit protein

synthesis in host cells

  • Mild to severe diarrheal disease
  • Low infectious dose
  • An array of other genes can contribute to

pathogenicity

  • More genes = more virulent

5 Cellfie of the Day: Bacterial toxin in the intestine of a child. Image credit: S. Schuller, Wellcome Images

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Case S Study dy: Germany

  • May-July 2011
  • 53 people died
  • Thousands sickened
  • New combination of genes

resulted in never-before-seen hybrid STEC

6

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Report rted S STEC I Infections in S South Dakota

7

South Dakota Department of Health

Five Year Median: 42 cases/year (+90%) Region Cases Rate Sioux Falls MSA 16 6.4 Rapid City MSA 7 5.2 Northeast 23 13.3 Southeast 17 15.1 Central 10 10.7 West 7 7.6 South Dakota 80 9.3 STEC Cases by Month of Onset

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SLIDE 8

Surface Water

Agricultural Input Urban Input Wildlife Input

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Research R h Rationa nale

  • South Dakota waterways are affected by fecal contamination
  • Study areas: Big Sioux River/Skunk Creek & Rapid Creek
  • Most E. coli are harmless commensal organisms
  • Lateral gene transfer can transform benign bacteria into pathogens
  • Aquatic environments harbor of Shiga toxin genes
  • E. coli levels provide an incomplete picture in relation to human

health and safety

9

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Path-STREAM Pathogenicity P Profiling: Shiga Toxins ns a and Relate ted

  • E. c

coli Attributes i identification Met ethod

  • Improved bacterial water quality

monitoring metric

  • Pathogenicity potential of ALL bacteria

present in a given water sample

  • Variety of virulence genes
  • Microbiological and molecular techniques
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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

stx1: 0% Health Implications: mild to moderate diarrhea

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

stx2: 55% Health Implications: moderate diarrhea

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

eae: 22% Health Implications: intestinal lesions

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

exhA: 20% Health Implications: bloody diarrhea

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

espP: 17% Health Implications: bloody diarrhea, kidney failure

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

eaeA: 3% Health Implications: intestinal lesions

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Rapid Creek

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

einV: 2.5% Health Implications: watery diarrhea, dysentery

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Rapid Creek

Temporal Distribution of Pathogenicity Genes

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  • 5

5 10 15 20 25 DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP November 2016 December 2016 January 2017 February 2017 March 2017 April 2017 May 2017 June 2017 July 2017 August 2017 September 2017 October 2017 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek

Temporal Distribution of Pathogenicity Genes

  • Avg. Temp 3.5°C

108 Gene Detections

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  • 5

5 10 15 20 25 DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP November 2016 December 2016 January 2017 February 2017 March 2017 April 2017 May 2017 June 2017 July 2017 August 2017 September 2017 October 2017 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek

Temporal Distribution of Pathogenicity Genes

  • Avg. Temp 3.5°C

108 Gene Detections

  • Avg. Temp 14.9°C

79 Gene Detections

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SLIDE 22
  • 5

5 10 15 20 25 DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP DC CL SP TP November 2016 December 2016 January 2017 February 2017 March 2017 April 2017 May 2017 June 2017 July 2017 August 2017 September 2017 October 2017 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek: : Ph Phase II

Temporal Distribution of Pathogenicity Genes

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Rapid Creek: : Ph Phase II

Dark Canyon Canyon Lake

  • E. St. Patrick St.

Wastewater Treatment Plant

43 44 44 69 Spatial Distribution of Pathogenicity Genes

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Rapid C Creek Summary

Genes Health Implications stx1, stx2 Mild to moderate diarrhea eae, eaeA Intestinal lesions einV Watery diarrhea, dysentery espP Bloody diarrhea exhA Bloody diarrhea

  • stx2 most commonly detected gene

(64%)

  • Appearance/disappearance of

particular genes may be related to temperature

  • Highest gene detection after the

wastewater treatment plant

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Health Risk

  • The combination of stx1, stx2, and eaeA are found in the most

pathogenic EHEC strains (including O157:H7)!

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One Organism: Rapid Creek

  • 255 Isolates
  • 22 Dark Canyon
  • 61 Canyon Lake
  • 86 E. St. Patrick St.
  • 86 Wastewater Treatment Plant

30% 1.5% 4%

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One Organism: Rapid Creek

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One Organism: Rapid Creek

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One Organism: Rapid Creek

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

Freshwater bacteria Opportunistic Pathogen: diarrhea, wound infections

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

Human commensal gut microbe Opportunistic Pathogen: diarrhea, UTI, sepsis

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

Nosocomial pathogens Bacteremia, respiratory infection, skin infection

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One One Or Organism sm: Rapi apid C Creek k Phylog

  • gen

enetic T Tree: ee: 16S Sequen encing

Gut microbe in mammals Opportunistic pathogen: wound infection, sepsis, UTI

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One Organism Summary

  • Multiple highly related enteric organisms harbor E. coli pathogenicity

genes

  • All organisms have been reported as stx carriers
  • Certain gene combinations were carried by multiple species of

bacteria; Lateral gene transfer

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Conclusions

stx1 stx2 eaeA exhA eae eaeγ1 einV espP toxB CNF1 Rapid Creek Phase 1     Phase 2      

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Conclusions

  • Virulence gene profiling of the bacterial community within surface

water provides a foundation for risk assessment associated with contacting fecally contaminated waterways

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Future Work

  • Whole genome sequencing of isolates
  • Functional genes?
  • May help identify the source,

donor/recipient relationship in LGT

  • Comparison to clinical isolates to

determine a link between disease and the environment

  • Expansion of Path-STREAM to include

more gene targets including those for antibiotic resistance and source tracking

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Future Work

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Acknowledgements

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Acknowledgements

Principal Investigators

  • Linda C. DeVeaux, Ph.D.
  • Lisa A. Kunza, Ph.D.
  • Kelsey Murray, Ph.D.

Undergraduate Research Associates

  • Sydney Stanley
  • Molly Erickson
  • Morgan Schaefer
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SLIDE 43

Questions?