1. C R E A T I N G A N D D E L I V E R I N G B E T T E R S O L U T I O N S E B A E N G I N E E R I N G C O N S U L T A N T S L T D. Presented by Gilles Wendling, Ph.D., P.Eng. Water Microbiology

2. Outline ! Why should we be concerned? -Brief history -What is the problem -Status in BC ! Outbreaks and pathogenes ! Bacteria and viruses -Definition -Characteristics ! Other waterborne -Algae and fungi ! Symptoms and illnesses ! What can we do about it? -treatment

3. Treatment: started ~100 years ago ! Began in early 1900’s in England (1908 in Chicago and Jersey City) -Continuous application of chlorine ! Waterborne disease such as typhoid fever was prevalent. -Major cities were suffering 100 or more typhoid deaths a year per 100,000 persons. ! 1910 – 1920: thousands of drinking water treatment plants initiated chlorine disinfection -The typhoid death rate fell simultaneously.

4. Source: Chlorine Chemistry Council

5. Treatment (cont’d) ! The number of typhoid fever cases is now insignificant -In developed countries -However, >250 000 children died in Bangladesh in 2000 due to waterborne diseases

6. Table 3. Waterborne Pathogens Legionella Vibrio (cholera) Salmonella (typhoid) Vibrio (noncholera) Yersinia ReovirusShigella Entamoeba histolyticaHepatitis ASalmonella (nontyphoid) Giarda lambliaEntero (poliomyelitis, coxsackie, echo, rotavirus) Escherichia coli Cryptosporidium parvumNorwalk-likeCampylobacter ProtozoaVirusesBacteria

7. Etiology of Waterborne Outbreaks in USA, 1971-1992 EtiologyOutbreaksCasses of IllnessHospitalized CasesDeaths Undetermined34182,4862530 Protozoan12743,952670 Bacterial * 9819,3517206 Viral5814,167850 Chemical ** 604,202456 TOTAL684164,1581,17012

8. Etiologic Agents Most Frequently Identified in Waterborne Outbreaks of Infectious Diseases in USA 1971-1992 Etiologic Agent OutbreaksCases of Illness Giardia lamblia 11826,733 Shigella 579,967 Norwalk-like virus 2410,908 Hepatitis A 29807 Campylobacter 135,257 Salmonella 122,370 Cryptosporidium parvum 717,194 All others * 234,243 TOTAL 28377,479 * Toxigenic E. coli, Yersinia, rotavirus, S. typhi, V. cholera and others

9. Causes of Waterborne Outbreaks in USA, 1971-92 Causes of OutbreakPercent of Outbreaks Community Water SystemsNon-Community Water Systems Contamination of distribution system29%7% Inadequate disinfection of unfiltered surface water 24%8% Inadequate disinfection of groundwater 14%30% Untreated groundwater 11%42% Inadequate filtration of surface water 11%1% Miscellaneous; unknown causes 5%6% Inadequate chemical feed 3%1% Untreated surface water 2%5% Inadequate filtration of groundwater 1%0% TOTAL 100%

13. Outline ! Why should we be concerned? ! Outbreaks and pathogenes ! Bacteria and viruses -Definition -Characteristics

14. How big are they?

15. http://micro.magnet.fsu.edu/cells/index.html Silt Sand Gravel Pore Size

16. Virus Fungi Pore Bacteria

17. Classification Until 1800s, two classes of living organisms ! Plants or animals (many people still have this perception) In 1866, Ernst Haeckel proposed a third kingdom, Protista, which included all microorganisms, such as bacteria, protozoans, and certain fungi and algae.

18. Classification Five-kingdom classification of living organisms ! H.R. Whittaker of Cornell University (1969) Alternative seven- and eight-kingdom classifications ! Recently ! Due to many factors (electron microscopy and gene sequencing, etc.) Now, the five-kingdom system is commonly used

19. Living Organisms: 5 groups

20. Bacteria A fundamental biological distinction is made between bacteria and all other organisms. ! genetic material (DNA and RNA) is NOT contained in a membrane-enclosed nucleus.

21. Nitrifying bacteria cross section Photo credit: Mary Ann Bruns, Center for Microbial Ecology, Michigan State

22. Bacteria Single-celled, with each cell carrying out all of the cell functions. Bacteria have three basic shapes: ! spherical, termed cocci (singular: coccus); ! rod shaped, termed bacilli (singular:bacillus); and ! spiral shaped. Bacteria can group themselves in distinctive ways. For example, cocci that form grape-like clusters are termed staphylococci and those that form chains are called streptococci Streptococcus pyogenes

23. Waterborne Bacteria Potential pathogens include Vibrio cholerae, Campylobacter jejuni, Salmonella species, and Escherichia coli (E. coli). Why pathogenes? ! secrete toxins that cause inflammation of the stomach and intestinal linings.

24. Color-enhanced scanning electron micrograph of Salmonella typhimurium (red) invading cultured human cells. Photo Source: Photo by Rocky Mountain Laboratories, NIAID, NIH with information from The National Institute of Allergy and Infectious Diseases

25. E-coli Universally used in standard tests as indicators of fecal contamination. E. coli strains vary widely in their pathogenicity. People tend to have a degree of immunity to local strains brought about by exposure (not visitors...! - tourista) More dangerous strains: i.e., E. coli 0157:H7, can cause hemorrhagic colitis that results in thousands of life- threatening illnesses and hundreds of deaths a year. (Although most are associated with food contamination, not contaminated drinking water)

26. http://www.microscopyconsulting.com/ E. coli O157:H7

27. Living Organisms

28. Protozoans ! Genetic material enclosed in a distinct membrane- covered nucleus. ! Protozoans have a flexible outer covering, rather than a semi-rigid cell wall like bacteria.

29. Protozans Play key roles in the environment: ! decay of organic matter to constituting a large portion of plankton ! Protozoans become dessicated easily and require damp or aquatic environments. They are generally much larger than bacteria ! some overlap between the largest bacteria and the smallest protozoans.

30. Protozoans – Important Feature Capable of producing cysts under adverse conditions: ! lack of moisture, food, or oxygen, or in the presence of toxic chemicals. ! In the case of parasitic protozoans, the cyst allows the organism to survive outside a host (days to weeks), which is crucial for those protozoans whose life cycle requires multiple hosts. Cysts present some challenges for water treatment because they are resistant to disinfection.

31. Protozoans and Cyst cycle giardia troph

32. Protozoa: Giardia Giardia lamblia ! possibly the most common cause of waterborne diarrheal disease in the north America ! commonly endemic in day-care centers ! more than 5% of adults and children are carriers who shed cysts in their feces, yet may have no symptoms. ! Infection is usually through cyst-contaminated water.

33. Giardia Cyst in Stool Photo Source: www-medlib.med.utah.edu/parasitology/glambim.htmlwww-medlib.med.utah.edu/parasitology/glambim.html

34. Protozoa: Cryptosporidium Cryptosporidium ! widespread in the natural environment ! probably of greater concern than Giardia -because its smaller size makes it harder to filter -it is more resistant to disinfection.

35. Other Protozoae Cyclospora and Microsporidia ! recently emerging protozoal pathogens. ! Cyclospora: -cysts are bigger than those of Cryptosporidium, must be mature before they are infectious. So, if cysts are shed from the body before maturation, the host can avoid infections. ! Microsporidia: -less known and it is likely that cases may be underreported.

36. Viruses: Definition ! Viruses fall into an entirely different category. ! In fact, they may not qualify to be called organisms.

37. Viruses: Definition ! All living organisms contain both DNA and RNA. Viruses may have DNA or RNA, but not both. ! Unlike true living organisms, their nucleic acids are inactive outside of host cells. -once they enter host cells, their nucleic acids become active, and they are able to replicate.

38. Photo Caption: This is a slide of a type B viral Hepatitis as shown in its chronic stage. Photo Source: Transplant Pathology at the University of Pittsburgh, tpis.upmc.edu/tpis.upmc.edu/ Photo Caption: This is a slide of a type B viral Hepatitis as shown in its chronic stage. Photo Source: Transplant Pathology at the University of Pittsburgh, tpis.upmc.edu/tpis.upmc.edu/

39. Viruses: Characteristics ! Viruses in general are much smaller than bacteria. -can be viewed only by electron microcroscopes. -there is some overlap in size between the largest viruses and the smallest bacteria. ! Their small size and resistance to environmental stress creates water treatment challenges. -viruses are less easily filtered by soil than bacteria, -viruses have been detected in groundwater that was absent of fecal indicator bacteria.

40. Viruses: Types ! Enteric viruses that have been associated with waterborne disease outbreaks -Hepatitis A virus, Hepatitis E virus, Norwalk and Norwalk-type viruses, Rotaviruses, and Enteroviruses. ( large group that includes Polioviruses).

41. Viruses: Testing and Treatment ! Difficult to test for the presence and concentration of viruses ! Water systems that use surface water or groundwater that is directly under the influence of surface water are required to use a combination of treatment options and disinfection that will result in a four-log (99.99 percent) reduction or inactivation of viruses.

42. Outline ! Why should we be concerned? ! Outbreaks and pathogenes ! Bacteria and viruses ! Other waterborne -Algae and fungi

43. Fungi ! have eucaryotic cells (genetic material in membrane-enclosed nucleus) ! no chlorophyll ! obtain nutrition by absorbing soluble substances across their cell walls Fungi that are multi-cellular fall into the Fungi kindgom, which includes mushrooms and most molds.

44. Living Organisms

45. Algae ! The term “algae” has no officially recognized biological meaning. ! Referred to any simple aquatic plant. Most biologists now use algae (singular: alga) to refer to organisms that have no true roots, stems, or leaves ! Have chlorophyll and are capable of photosynthesis.

46. Mixed bloom of Dinophysis acuta and D. norvegica co-occurring with a bloom of Ceratium furca

47. Living Organisms

48. Algae ! Largely viewed mainly as the cause of taste, odor, and color problems ! Some algae (cyanobacteria Microcystis, Cylindrospermopsis, and Anabaena) have recently come under more scrutiny in the U.S. for their health effects (linked to nerve and liver damage).

49. Algae ! Algae and their toxins are currently unregulated by the Safe Drinking Water Act. However, they are included on EPA’s Drinking Water Candidate Contaminant List (CCL). ! Australia has a regulatory program in place for cyanotoxins and the World Health Organization has a 1.0 microgram per liter guideline for microcystin.

50. Outline ! Why should we be concerned? ! Outbreaks and pathogenes ! Bacteria and viruses ! Other waterborne ! Symptoms and illnesses

51. Illnesses Associated with Waterborne Pathogens Bacteria and protozoa generally induce gastrointestinal disorders with a wide range of severity. Bacteria also cause life-threatening diseases such as typhoid and cholera. Viruses cause serious diseases such as aseptic meningitis, encephalitis, poliomyelitis, hepatitis, myocarditis and diabetes.

52. Illnesses Associated with Waterborne Pathogens ! Severity of illness ranges from mild gastrointestinal upset, fever and vomiting, and intermittent diarrhea to chronic diarrhea, dehydration, liver damage, acute respiratory illness, adverse neurological effects, depressed immune systems and death. ! Most healthy individuals in the general population usually experience only mild gastroenteritis that is easily controlled and of short duration. ! Certain segments of the population are especially vulnerable to acute illness (morbidity) and can exhibit high death (mortality) rates. -pregnant women, infants, the elderly and those whose immune systems are compromised by cancer, AIDS

53. Outline ! Why should we be concerned? ! Outbreaks and pathogenes ! Bacteria and viruses ! Other waterborne ! Symptoms and illnesses ! What can we do about it? -treatment

54. Drinking Water Treatment: Goals ! 99% removal of Cryptosporidium oocysts (2 log) ! 99,9% removal of Giardia cysts (3 log) ! 99,99% removal of viruses (4 log) But these goals are intimately related to the credit given to conventional treatments (otherwise an economical crisis would be created) The log removal credits are: ! Specific to a technology or process ! Specific to each organism

55. Drinking Water Treatment : Methods Two major types of treatments: ! Filtration ! Disinfection Filtration and UV disinfection are more efficient for protozoa Chemical disinfections are more efficient for viruses

56. Drinking Water Treatment: Filtration ! Turbidity is an indicator of the overall water quality ! Turbidity is use to make sure disinfection will be efficient ! Turbidity may also be used to follow treatment performances: -Chemically-assisted filtration (conventional treatment or direct filtration), slow sand filtration and diatomaceous earth filtration are given general process log removal credits based on the turbidity of the filtered water ! Variation of treated water turbidity is more important than its actual value

57. Drinking Water Treatment: Disinfection ! Many disinfectant can be used and their efficiency varies a lot depending on the target organism ! The most commonly used are: -Free chlorine -Chlorine dioxide -Ozone -Chloramines (monochloramines) ! Depends on residual concentration, contact time, temperature and pH

58. CT values at 5ºC for 1 log removal: Drinking Water Treatment : Contact Time (mg*min/L)E. coliGiardiaCrypto.Viruses Free chlorine (0,5 mg/L, pH 6-7) 0,02 – 0,03 32 - 47-0,01 – 2,0 Chlorine dioxide 0,4 – 0,88,74240,1 – 3,4 Monochloramines (pH 8-9) 95 - 180735-350 - 3500 Ozone 0,020,6321,20,003 – 0,3

59. Drinking Water Treatment: membrane filtration and UV UV disinfection: Example of UV dose required: No residual after the reactor: another disinfectant must be used for distribution If an alarm triggers, water must be stopped because there is no residual (mJ/cm²)E. coliGiardiaCrypto.Viruses UV dose (1 log) 1,5 – 4,42,12,54,1 - 30 UV dose (3 log) 4,1 – 7,3111214 - 90

60. Final note…Our knowledge is very relative Number of generation since Christ: 100 The earth is flat…till Galileo: only 25 generations ago! First dissections on cadavres ~ Da Vinci (1452- 1519)..25 generations ago Discovery of bacteria: ! Related to discovery of microscope ! Recent big step with use of electronic microscope

61. Final note…Our knowledge is very relative ……..We still have a lot to discover ! Nov 2004: discovery of bacteria 100x larger that bacteria known to date!!!

62. Recommendations Groundwater can be a very safe resource ! Proper installation and sealing of well ! High wellhead standard ! Source protection

63. Credits http://c3.org/chlorine_knowledge_center/whitepapercl.html#Water http://www.whoi.edu/redtide/rtphotos/rtphotos.html http://micro.magnet.fsu.edu/cells/index.html Hughes, J.M. 1993. Infectious Diseases Transmitted by Drinking Water in the United States: Perspectives of the Centers for Disease Control and Prevention. Pp. 11-16. Safety of Water Disinfection: Balancing Chemical & Microbial Risks. ILSI. Craun, G.F., Ed. and multiple contributors. 1993. Safety of Water Disinfection: Balancing Chemical & Microbial Risks. ILSI. Craun, G.F., and multiple contributors. 1996. Water Quality in Latin America: Balancing the Microbial and Chemical Risks in Drinking Water Disinfection. Proceedings of the Regional Symposium on Water Quality, sponsored by International Life Sciences Institute, Argentina. Craun, G.F., Bull, R.J., Clark, R.M., Doull, J., Grabow, W., Marsh, G.M., Okun, D.A., Regli, S., Sobsey, M.D., and Symons, J.M. 1994. Balancing Chemical and Microbial Risks of Drinking Water Disinfection, Part I. Benefits and Potential Risks. Pp. 192-199. J Water SRTÑAgua. 43:4. Craun, G.F., Bull, R.J., Clark, R.M., Doull, J., Grabow, W., Marsh, G.M., Okun, D.A., Regli, S., Sobsey, M.D., and Symons, J.M. 1994. Balancing Chemical and Microbial Risks of Drinking Water Disinfection, Part II. Managing the Risks. Pp. 207-218. J Water SRTÑAgua. 43:5. Craun, G.F. 1994. Waterborne Disease in the United States. 1996. Pp. 55-77. Water Quality in Latin America. Pan American Health Organization. Chain-Wen Wang and Craig Mains

64. C R E A T I N G A N D D E L I V E R I N G B E T T E R S O L U T I O N S E B A E N G I N E E R I N G C O N S U L T A N T S L T D. Thank You