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Applied and Environmental Microbiology, June 2005, p. 3163-3170, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.3163-3170.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Validity of the Indicator Organism Paradigm for Pathogen Reduction in Reclaimed Water and Public Health Protection{dagger}

Valerie J. Harwood,1* Audrey D. Levine,2 Troy M. Scott,3 Vasanta Chivukula,1 Jerzy Lukasik,3 Samuel R. Farrah,4 and Joan B. Rose5

Department of Biology, SCA 110, University of South Florida, 4202 E. Fowler Ave., Tampa, Florida 33620,1 Department of Civil and Environmental Engineering, ENB 118, University of South Florida, 4202 E. Fowler Ave., Tampa, Florida 33620,2 Biological Consulting Services of N. Florida, Inc., 4641 N.W. 6th Street, Suite A, Gainesville, Florida 32609,3 Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida 32611,4 Department of Fisheries and Wildlife and Crop and Soil Sciences, 13 Natural Resources Building, Michigan State University, East Lansing, Michigan 488245

Received 27 September 2004/ Accepted 20 December 2004

The validity of using indicator organisms (total and fecal coliforms, enterococci, Clostridium perfringens, and F-specific coliphages) to predict the presence or absence of pathogens (infectious enteric viruses, Cryptosporidium, and Giardia) was tested at six wastewater reclamation facilities. Multiple samplings conducted at each facility over a 1-year period. Larger sample volumes for indicators (0.2 to 0.4 liters) and pathogens (30 to 100 liters) resulted in more sensitive detection limits than are typical of routine monitoring. Microorganisms were detected in disinfected effluent samples at the following frequencies: total coliforms, 63%; fecal coliforms, 27%; enterococci, 27%; C. perfringens, 61%; F-specific coliphages, ~40%; and enteric viruses, 31%. Cryptosporidium oocysts and Giardia cysts were detected in 70% and 80%, respectively, of reclaimed water samples. Viable Cryptosporidium, based on cell culture infectivity assays, was detected in 20% of the reclaimed water samples. No strong correlation was found for any indicator-pathogen combination. When data for all indicators were tested using discriminant analysis, the presence/absence patterns for Giardia cysts, Cryptosporidium oocysts, infectious Cryptosporidium, and infectious enteric viruses were predicted for over 71% of disinfected effluents. The failure of measurements of single indicator organism to correlate with pathogens suggests that public health is not adequately protected by simple monitoring schemes based on detection of a single indicator, particularly at the detection limits routinely employed. Monitoring a suite of indicator organisms in reclaimed effluent is more likely to be predictive of the presence of certain pathogens, and a need for additional pathogen monitoring in reclaimed water in order to protect public health is suggested by this study.


* Corresponding author. Mailing address: Department of Biology, SCA 110, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620. Phone: (813) 974-1524. Fax: (813) 974-3263. E-mail: vharwood{at}cas.usf.edu.

{dagger} Florida Agricultural Experiment Station Journal Series no. R10718.


Applied and Environmental Microbiology, June 2005, p. 3163-3170, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.3163-3170.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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