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Applied and Environmental Microbiology, February 2006, p. 1604-1612, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1604-1612.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Multitiered Approach Using Quantitative PCR To Track Sources of Fecal Pollution Affecting Santa Monica Bay, California

Rachel T. Noble,1* John F. Griffith,2 A. Denene Blackwood,1 Jed A. Fuhrman,3 Jason B. Gregory,1 Ximena Hernandez,3 Xiaolin Liang,3 Angie A. Bera,4 and Kenneth Schiff2

University of North Carolina—Chapel Hill, Chapel Hill, North Carolina,1 Southern California Coastal Water Research Project, Westminster,2 University of Southern California, Los Angeles,3 Santa Monica Baykeeper, Santa Monica, California4

Received 16 June 2005/ Accepted 16 November 2005

The ubiquity of fecal indicator bacteria such as Escherichia coli and Enterococcus spp. in urban environments makes tracking of fecal contamination extremely challenging. A multitiered approach was used to assess sources of fecal pollution in Ballona Creek, an urban watershed that drains to the Santa Monica Bay (SMB) near Los Angeles, Calif. A mass-based design at six main-stem sites and four major tributaries over a 6-h period was used (i) to assess the flux of Enterococcus spp. and E. coli by using culture-based methods (tier 1); (ii) to assess levels of Enterococcus spp. by using quantitative PCR and to detect and/or quantify additional markers of human fecal contamination, including a human-specific Bacteroides sp. marker and enterovirus, using quantitative reverse transcriptase PCR (tier 2); and (iii) to assess the specific types of enterovirus genomes found via sequence analysis (tier 3). Sources of fecal indicator bacteria were ubiquitous, and concentrations were high, throughout Ballona Creek, with no single tributary dominating fecal inputs. The flux of Enterococcus spp. and E. coli averaged 109 to 1010 cells h–1 and was as high at the head of the watershed as at the mouth prior to discharge into the SMB. In addition, a signal for the human-specific Bacteroides marker was consistently detected: 86% of the samples taken over the extent during the study period tested positive. Enteroviruses were quantifiable in 14 of 36 samples (39%), with the highest concentrations at the site furthest upstream (Cochran). These results indicated the power of using multiple approaches to assess and quantify fecal contamination in freshwater conduits to high-use, high-priority recreational swimming areas.

* Corresponding author. Mailing address: UNC Chapel Hill Institute of Marine Sciences, 3431 Arendell St., Morehead City, NC 28557. Phone: (252) 726-6841, ext. 150. Fax: (252) 726-2426. E-mail: rtnoble{at}email.unc.edu.

Applied and Environmental Microbiology, February 2006, p. 1604-1612, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1604-1612.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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