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Giardia Cyst and Cryptosporidium Oocyst Survival in Watersheds and Factors Affecting Inactivation [Project #151]

Ordering Information:
ORDER NUMBER: 90761
DATE AVAILABLE: Summer 1999

Printed Report
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PRINCIPAL INVESTIGATORS:
S.A. Sattar, C. Chauret, V.S. Springthorpe, D. Battigelli, Morteza Abbaszadegan, and Mark LeChevallier

OBJECTIVES:
The overall objective of this study was to understand the environmental factors that influence the survival or inactivation of protozoan parasite cysts and oocysts, and their susceptibility to subsequent disinfection. This refers to the physical, chemical and biological factors prevailing in natural or managed water environments, and to the stressors that may be encountered or applied during in-plant treatment.

BACKGROUND:
Cysts/oocysts are resistant forms of protozoa shed from infected hosts, and subsequently discharged into receiving waters through various point (e.g., sewage effluent) and nonpoint (e.g., land run-off) sources. The resistance of these forms to many stressors, including the disinfectants used in water treatment, makes them a potential hazard for water consumers and a challenge for the water industry. Understanding the factors that affect inactivation of such pathogens in watersheds may encourage better management practices.

HIGHLIGHTS:
The physical factors that were most influential in parasite decline were water temperature, and sunlight or blacklight. Within the confines of the study, the chemical composition of water did not play a large role in oocyst decline. The greatest decline observed was attributable to biological components, between
0.2 and 5 μm in diameter, present in the natural waters tested. However, results varied between watersheds and seasons. Generally, Giardia muris appeared more sensitive to stressors than did Cryptosporidium. Aged or stressed oocysts were no less resistant to disinfection than those freshly shed.

APPROACH:
Parasite survival experiments were performed in standard hard water as well as in natural water samples from different watersheds in Canada and the U.S. Relevant controls were included in all experiments. Most experiments were conducted in vitro in small polypropylene tubes, but some experiments were conducted in situ using flow-through tanks and dialysis cassettes. Physicochemical environmental stressors were applied separately and in various combinations. Early indications that biological components in water can affect parasite inactivation were followed by more detailed investigations of this phenomenon. Parasite survival was assessed as a function of time by excystation assays and counts of total cyst or oocyst numbers. Aged or stressed oocysts were used, in comparison with fresh ones, for disinfection experiments.

RESULTS/FINDINGS:

Physicochemical Stressors
In general, Giardia muris cysts were very sensitive to stressors at ambient temperatures. Therefore, most experiments used only Cryptosporidium. Increased temperature had the most influence on parasite decline in standard hard water. Sunlight or long wavelength UV (blacklight) was also detrimental. Water hardness, pH, and redox had little effect in the ranges tested. Sand shearing and freezing/thawing were more damaging. The anion and metal content of the water played no role in oocyst decline in vitro. However, the influence of chemicals in situ in the aquatic environment may be more prominent and would require further investigation.

Biological Factors
While biological activity emerged as perhaps the most prominent stressor in some water sources, the effects were not observed across the board. This may be attributable to different microbial populations between water sources and seasons. Concentration of the indigenous biota, between 0.2 and 5 μm in diameter, can serve to pinpoint the importance of microbial antagonistic activity in any watershed. Tests with individual bacterial isolates or groups of isolates failed to demonstrate significant oocyst inactivation. This suggests that the organism(s) responsible may not be cultivable or may act in complex consortia.

Disinfection
Aged or stressed oocysts were no less resistant to disinfection than were those freshly shed.

IMPACT:
This study is a building block upon which models, essential to justify good watershed management, could be designed to predict the inactivation of Cryptosporidium oocysts and Giardia cysts. Findings of this study show that Cryptosporidium oocysts survive well and thus travel far from the pollution source. This emphasizes the importance of applying best management practices throughout the watershed.

Aged/stressed oocysts were as resistant to disinfection as freshly excreted ones. Thus, it is important for utilities to implement effective removal and disinfection practices. Accordingly, the data generated in previous studies on disinfection kinetics (CT values, etc.) of fresh oocysts must be regarded as accurate for evaluating CT values of environmental isolates.

PARTICIPATING UTILITIES:

  • American Water Works Services Company, Inc., East St. Louis, Ill.
  • Regional Municipality of Ottawa-Carleton, Ottawa, Ont., Canada
  • City of Cornwall, Cornwall, Ont., Canada
  • Regional Municipality of Waterloo, Kitchener-Waterloo, Ont., Canada


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