1. Ground Water Risks & Microbes Joan B. Rose 517-432-4412 rosejo@msu.edu

2. Groundwater Use in Michigan 2001 Total 513.36 MGD

4. Water Quality and Fecal Indicators Criteria and Standards / 100 mL FOR RECREATIONAL WATERS E.coli - 235 CFU US EPA - 300 CFU MDEQ Enterococci - 61 CFU US EPA Clostridium - 50 CFU Hawaii FOR DRINKING AND GROUND WATER <1 Total coliform bacteria and E.coli; Enterococci or coliphage No detectable pathogens (viruses and parasites) Parasite detection in ground water is viewed as “ground water under the influence of surface waters” and should be filtered for drinking purposes.

5. E. coli Standard in Michigan Found in the gastrointestinal tract of mammals Member of Fecal Coliform group Possess the enzyme β-glucuronidase Presence indicates fecal contamination

6. Enterococci Sub group of streptococci Used in testing water quality EPA recommended and greater association with recreational disease Some can readily colonize skin and mucous membranes

7. Clostridium perfringens Anaerobe Spore former Produces toxins

8. Coliphage Virus Needs bacteria’s machinary (E.coli) to replicate Indicates fecal contamination

9. Types of Waterborne Pathogens Viruses Bacteria Parasites THE DISEASES: diarrhea, respiratory illness, liver damage, kidney failure, heart disease, cancer, nervous system disorders; birth defects, death.

10. Chief Waterborne Microorganisms Viruses –rotavirus –coxsackievirus –echovirus –calicivirus –norovirus –Hepatitis A and E Bacteria –E.coli –Salmonella spp. –Shigella spp. –Aeromonas hydrophila –Campylobacter jejuni Protozoa –Cryptosporidium parvum –Giardia lamblia

11. OVER 100 Viruses found in sewage:Adenoviruses, respiratory and enteric, higher numbers, greater resistance to uv disinfection. Coxsackie viruses, myocarditis and viral meningitis. Identification of Cancer causing viruses Polyomaviruses in Wastewater. SMALL BIO- NANO PARTICLES LONG-TERM SURVIVAL HIGHLY INFECTIOUS

13. Figure 3

14. Figure 6

18. WATERBORNE OUTBREAKS IN THE U.S. Polluted groundwater a major risk Small systems Unknown and emerging contaminants

19. Waterborne Disease in the U.S. 1993-1994: 14 outbreaks, 37% groundwater 1995-1996: 22 outbreaks, 50% groundwater 1997-1998: 17 outbreaks, 88% groundwater Viral contamination detected in 40% of groundwater sources tested

20. Groundwater Monitoring Study transport and survival of pathogenic microbes in groundwater Monitor for pathogens using culture and PCR (molecular methods)

21. Polymerase Chain Reaction (PCR) Developed in 1985 by Kary Mullis Dr. Mullis received the Nobel Prize in Chemistry in 1993 PCR is considered as one of the most important discoveries in molecular biology

22. What is PCR? Enzymatic reaction that makes many copies of DNA from single molecule 2 n copies of DNA from single molecule where n = No. of cycles So, 35 cycles of PCR would yield 2 35 copies of DNA

24. Source Tracking at MSU Recently developed method to track a Human marker in Enterococci. 107/109 samples from human sewage and septic tanks were positive. 0/80 samples from cattle, swine, bird, fecal samples and lagoons were positive.

25. Human Pollution Marker Validation Sourceesp marker + Sewage67/67 Septic tank14/16 Poultry0/8 Swine0/12 Cattle0/35 Canada Geese0/12 Seagull0/28 Pelican0/7 Wild Birds0/10 Scott et al., 2005. Env. Sci. Technol., 39: 283-287

27. Enteric viruses In a National Survey approximately 8% of ground waters in U.S. were positive for cultivatible viruses while 30% were positive by Molecular methods (PCR). PCR is being used to detect of risk for Viruses which can not be easily grown in cell culture.

28. Sampling locations + for esp Marker Pathogen and Indicator Sampling in Michigan Waters + for Enteric Viruses

29. esp human marker in Great Lakes recreational beach waters Silver Beach, MI Samplin g time Number of swimmers Mean turbidity (NTU) [SD] positive for esp marker Morning 17 3.2 [3.8] 5/168 3% (n=276) Afternoo n 805 3.7 [3.7] 4/88 4.5% (n=138) Total 8229/256

31. Studies in Wisconsin have shown statistical Association between diarrhea in children and Virus infection and Septic tank density. No evidence of well contamination based on coliforms. Virus infections associated in a 640-acre section. Bacterial infections in a 40-acre section. Unknown infections associated with Enterococci contamination. Borchardt et al. May 2003 Envir. Hlth Perspec vol 111

32. Groundwater Risks Lessons Learned Walkerton, Ontario Outbreak (occured In small community Using Ground water). Source: Application of Animal Waste/Manure Monitoring and Disinfection not addressed. 2300 CASES 7 DEATHS 27 CASES of HUS 5 years later community still suffering.

33. E.coli 0157:H7 Enteropathogenic strain of E. coli Serious waterborne outbreaks 243 cases, 32 hospitalizations, 4 deaths Water main repair with sewage overflow contamination, 1987 Groundwater supplying Fairgrounds in NY 750 cases, 2 deaths, 1999 Walkerton, groundwater, ~1000 cases, 6 deaths, 2000

34. E.coli 0157:H7 Health Effects Children and the elderly at greatest risk Severe bloody diarrhea Hemolytic uremic syndrome Kidney failure Death

35.  The WHO has classified H. pylori as a Class I carcinogen because of the association of H. pylori and gastric malignancies.  German group,Rolle-Kampczyk et al. ( 2004) found a significant correlation between well water contaminated with H. pylori detection by PCR and colonization status in humans.  Water supplies contaminated with fecal material may be a potential source of H. pylori transmission (Hulten et al., 1996).

37. Analysis of VacA gene of H. pylori Vs Samples No. of H. pylori / 50 ml samples Raw water from waste water treatment plant at different period of time

38. Campy and new emerging bacteria associated with ground water Campylobacter jejuni is a major cause of human bacterial enteritis. Source of transmission of C. jejuni to humans occurs via contaminated water, poultry, shellfish and milk. Arcobacter spp have been associated with cases of human enteritis and abortion in livestock. Because of their phylogenetic proximity, transmission mechanisms that have been described for C. jejuni may be applicable to Helicobacter and Archobacter spp (Wesley V.I.,1997 )

39. Campy and new emerging bacteria associated with ground water Campylobacter jejuni is a major cause of human bacterial enteritis. Source of transmission of C. jejuni to humans occurs via contaminated water, poultry, shellfish and milk. H. pylori is the most common chronic bacterial infection to occur in humans which leads to gastric cancer. Arcobacter spp have been associated with cases of human enteritis and abortion in livestock. Because of their phylogenetic proximity, transmission mechanisms that have been described for C. jejuni may be applicable to Helicobacter and Archobacter spp (Wesley V.I.,1997 )

40. “DON’T DRINK THE WATER— DRINK THE WINE” THE SOUTH BASS ISLAND OUTBREAK of 2004

41. Washington-area hotel closes for cleaning after norovirus sickens dozens of guests The Associated Press Published: March 2, 2007 ARLINGTON, Virginia: A hotel near a Washington, D.C., airport was closed for cleaning after as many as 150 guests were sickened by the highly contagious norovirus, hotel and county health officials said. FAIRFAX COUNTY Senior Community Hit by Possible Norovirus By Leef SmithLeef Smith Washington Post Staff Writer Saturday, March 10, 2007; Page B02 By kgw.com Staff In 2006, 145 outbreaks sickened 5000 people in Michigan nursing homes, restaurants, college campuses, a prison, and summer camp

42. Ohio blames groundwater for Lake Erie island outbreak Tuesday, February 22, 2005 ASSOCIATED PRESS TOLEDO, Ohio -- Widespread groundwater contamination on a Lake Erie resort island was the likely source of illnesses that sickened hundreds last summer, the Ohio health department said Tuesday. Several sources, including septic tanks, have tainted the South Bass Island's groundwater over a long period, and the contamination may have been worsened last summer because of a season of heavy rains, a health department report said. The outbreak of gastrointestinal illness sickened about 1,400 tourists and residents, ending the tourist season early for many businesses. MSU assisted with the investigation Identified virus contamination and potentially a new and emerging bacteria

44. South Bass Island Lake Erie Sources septic Tanks & sewage discharges Massive Ground Water and SurfaceWater Contamination

45. Outcomes of Outbreak Investigation On Feb 23, 2005, OH Health Dept. announced that the outbreak was caused by consumption of contaminated water heavy rains in May 2004 (the fifth wettest May in OH history) may have worsened the contamination the island's porous soil makes it easier for contaminants such as bird droppings and motor oil to wash into the bedrock especially during heavy rains rising and falling lake levels shallow wells, min state requirement- 25 ft

46. Needed Ground Water Studies Building Data bases: more monitoring, new indicators, virus testing. To Date we have examined 10 wells (30% positive for Coliphage) Tracer studies: Impact of Septic Tanks Understanding Ground water and surface water interactions

47. Tracer Studies In viral tracer studies, a nonpathogenic bacterial virus is seeded into a septic tank or injection well at high concentration, and surface or groundwaters monitored for viral appearance as a function of time.

48. TRACER STUDIES Used Bacteriophage Vibrio Phage PRD1 Salmonella Phage MS2, E.coli Phage Bacteriophage can be grown to high titer in the laboratory (10 million). A single plaque forming unit can be detected in 10 to 100ml using specific bacterial host and the agar overlay method

51. What we have learned from viral tracer Studies 1.Both septic tanks and injection wells Rapidly contaminated surface water (often In as little as 3 hr). 2. Rates of migration of wastewater ranged From 150 m/h 3. These results have shaped future waste Disposal policy in the Keys

52. MICHIGAN NEEDS A PATHOGEN AND SOURCE TRACKING DATA BASE MICHIGAN NEEDS BETTER APPROACH FOR HEALTH REPORTING MICHIGAN NEEDS TO BETTER UNDERSTAND THE RELATIONSHIP BETWEEN LANDUSE, CLIMATE (RAIN) AND POLLUTION MICHIGAN NEEDS TO RANK THE IMPACTS IN THE BEACHSHEDS AND WATER QUALITY AT THE BEACH. MICHIGAN NEEDS TO ADDRESS WASTEWATER INFRASTRUCTURE IMPROVEMENT