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

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 • Total fecal • E. coli coliforms

Escherichia c coli : A Geneti tically Diverse Species of B Bacteri ria • Routine testing provides a snapshot microbial abundance and content, but not pathogenic Commensal to Highly adapted profile of the bacteria Environmental and mammalian strains that allow for naturalized intestinal tracts; a broad spectrum of • The difference lies Probiotics human disease within the genes (DNA) 3

The Promiscuity ty of B Bacteri ria 4

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

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

Report rted S STEC I Infections in S South Dakota STEC Cases by Month of Onset 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 Five Year Median: 42 cases/year (+90%) South Dakota Department of Health 7

Agricultural Input Urban Input Surface Water Wildlife Input

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

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

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon Wastewater Treatment Plant

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon stx1 : 0% Wastewater Health Implications: mild to Treatment Plant moderate diarrhea

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon stx2 : 55% Wastewater Health Implications: Treatment Plant moderate diarrhea

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon eae : 22% Wastewater Health Implications: Treatment Plant intestinal lesions

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon exhA : 20% Wastewater Health Implications: bloody Treatment Plant diarrhea

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon espP : 17% Wastewater Health Implications: bloody Treatment Plant diarrhea, kidney failure

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon eaeA: 3% Wastewater Health Implications: Treatment Plant intestinal lesions

Rapid Creek E. St. Patrick St. Canyon Lake Dark Canyon einV : 2.5% Wastewater Health Implications: watery Treatment Plant diarrhea, dysentery

Rapid Creek Temporal Distribution of Pathogenicity Genes

Rapid Creek 25 Temporal Distribution of Pathogenicity Genes 20 15 Avg. Temp 3.5°C 10 108 Gene Detections 5 0 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 December January February March April May June July August September October 2016 2016 2017 2017 2017 2017 2017 2017 2017 2017 2017 2017 -5 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek 25 Temporal Distribution of Pathogenicity Genes 20 15 Avg. Temp 3.5°C Avg. Temp 14.9°C 10 108 Gene 79 Gene Detections Detections 5 0 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 December January February March April May June July August September October 2016 2016 2017 2017 2017 2017 2017 2017 2017 2017 2017 2017 -5 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek: : Ph Phase II 25 Temporal Distribution of Pathogenicity Genes 20 15 10 5 0 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 December January February March April May June July August September October 2016 2016 2017 2017 2017 2017 2017 2017 2017 2017 2017 2017 -5 stx1 stx2 eae eaeA einV espP exhA Temp (°C)

Rapid Creek: : Ph Phase II Spatial Distribution of Pathogenicity Genes 44 43 E. St. Patrick St. Canyon Lake 44 Dark Canyon 69 Wastewater Treatment Plant

Rapid C Creek Summary • stx2 most commonly detected gene Genes Health Implications (64%) stx1 , stx2 Mild to moderate diarrhea • Appearance/disappearance of eae , eaeA Intestinal lesions particular genes may be related to einV Watery diarrhea, dysentery temperature espP Bloody diarrhea • Highest gene detection after the exhA Bloody diarrhea wastewater treatment plant

Health Risk • The combination of stx1 , stx2 , and eaeA are found in the most pathogenic EHEC strains (including O157:H7)!

One Organism: Rapid Creek • 255 Isolates 30% • 22 Dark Canyon • 61 Canyon Lake • 86 E. St. Patrick St. • 86 Wastewater Treatment Plant 4% 1.5%

One Organism: Rapid Creek

One Organism: Rapid Creek

One Organism: Rapid Creek

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing Freshwater bacteria Opportunistic Pathogen: diarrhea, wound infections

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing Human commensal gut microbe Opportunistic Pathogen: diarrhea, UTI, sepsis

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing Nosocomial pathogens Bacteremia, respiratory infection, skin infection

One One Or Organism sm: Rapi apid C Creek k Phylog ogen enetic T Tree: ee: 16S Sequen encing Gut microbe in mammals Opportunistic pathogen: wound infection, sepsis, UTI

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

Conclusions stx 1 stx 2 eaeA exhA eae eaeγ1 einV espP toxB CNF1 Rapid Creek     Phase 1       Phase 2

Conclusions • Virulence gene profiling of the bacterial community within surface water provides a foundation for risk assessment associated with contacting fecally contaminated waterways

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

Future Work

Acknowledgements

Acknowledgements Principal Investigators Undergraduate Research Associates • Linda C. DeVeaux, Ph.D. • Sydney Stanley • Lisa A. Kunza, Ph.D. • Molly Erickson • Morgan Schaefer • Kelsey Murray, Ph.D.

Questions?