Microbial Source Tracking at Arroyo Burro Beach Jared Ervin Holden Lab, UCSB Sept. 20 th , 2012
Arroyo Burro Microbial Source Tracking � Part of a larger multi-lab project called the Source Identification Protocol Project (SIPP) � Funded through California Proposition 84 through the Clean Beaches Initiative � SIPP project goal is to identify and eliminate sources of fecal pollution at California beaches � Protect the health of beach users
Arroyo Burro (Hendry’s) Beach � #7 on Heal the Bay’s top ten beach bummer list in 2011, #9 in 2007 � F grade on 4 of the last 6 annual report cards � Grades are based on County of Santa Barbara weekly surf zone water quality monitoring *From Heal the Bay’s 2011 Annual Beach Report Card
Fecal Indicator Bacteria (FIB) � Monitoring required during the AB411 Period: ▫ May 1 st thru October 31 st ▫ Sample taken each week at each beach ▫ 16 beaches monitored in Santa Barbara County � Single sample limits set by the State of California ▫ Total Coliforms: 10,000 “cells” per 100mL ▫ Fecal Coliforms/E. coli: 400 “cells” per 100mL ▫ Enterococcus : 104 “cells” per 100mL ▫ TC>1,000 & FC:TC ratio greater than 0.1
*From: County of Santa Barbara weekly surf zone sampling
Arroyo Burro FIB Example 10-24-11 10-26-11 Limit Total Coliforms 882 122 10,000 Fecal Coliforms 697 31 400 Enterococcus 487 <10 104 TC>1,000 FC/TC Ratio N/A N/A 0.1 Beach Status Warning Open *From: County of Santa Barbara weekly surf zone sampling
Fecal Indicator Bacteria (FIB) *From: Cao et. al. 2011, Microbial Source Tracking
Why measure FIB? *From: Cao et. al. 2011, Microbial Source Tracking
FIB Limitations � Occur naturally in the environment � Not specific to sources of fecal contamination FIB Culturing FIB � Other tools needed to identify and locate sources of fecal pollution
Microbial Source Tracking (MST) 1. What are the source(s) of contamination? ▫ Source Identification ▫ Characterizing the origin of the bacteria (e.g., humans, birds, or livestock) 2. Where is the contamination originating? ▫ Source Tracking ▫ Following bacterial signal back to its source (e.g., a specific storm drain, campground, or leaking sewage pipe)
MST Tools � FIB culturing � DNA based analyses ▫ qPCR targeting specific fecal sources ▫ Whole bacterial community analysis � Other methods ▫ Dye studies and televising of sewers ▫ Detection of caffeine and cotinine
Host-Specific DNA Markers Cow Specific Human Specific Bird Specific Bacterial DNA Bacterial DNA Bacterial DNA
How DNA-based Tools Work: qPCR of Human Bacteroides marker 1 copy 1. Sample the water 2 copies 4 copies 8 copies 20x = 1 million copies 4. Amplify target DNA (qPCR) 5. Quantify human Bacteroides in 3. Extract DNA water sample 2. Capture bacteria
SIPP – Methods Evaluation � 43 novel microbial source tracking methods tested (mostly DNA-based) � Tested at 25 different labs ▫ 18 methods tested at UCSB � 12 sources of fecal contamination ▫ Human, septage, sewage, chicken, cow, deer, dog, goose, gull, horse, pig & pigeon � Most effective methods used in field studies.
SIPP – Field Studies � SCCWRP ▫ Doheny State Beach, Dana Point, CA � Stanford ▫ Cowell Beach, Santa Cruz, CA � UCLA ▫ Topanga State Beach, Malibu, CA � UCSB ▫ Arroyo Burro Beach, Santa Barbara, CA
Arroyo Burro Source Tracking � Analysis of historical data ▫ Sampling results from City & County ▫ Previous work done at Holden Lab & City � Watershed reconnaissance ▫ Investigation of all potential sources � Field sampling ▫ 10 weeks of sampling during summer 2012 ▫ Samples collected at upstream locations, in the lagoon and across the beach ▫ Sand and wrack samples also collected
Potential FIB Sources � At the Beach ▫ Birds & Dogs ▫ Sand & Wrack (i.e. kelp) � In the Lagoon ▫ Birds ▫ Humans � Upstream ▫ Humans ▫ Horses *Human sources may include homeless, leaking sanitary sewers and failing septic systems.
Sampling Design � At the Beach ▫ 5 locations across the surf zone ▫ Sand and Wrack samples � In the Lagoon ▫ 3 locations in the lagoon ▫ Creeks and lagoon outlet � Upstream ▫ 5 locations along Arroyo Burro and Las Positas Creeks
Current Project Status � Summer 2012 Field Sampling completed ▫ 178 water samples collected ▫ 30 sand and 30 wrack samples ▫ All samples filtered and frozen for DNA analyses � FIB and field data are being analyzed � Beach survey, photo and video data are being compiled and analyzed
Source Tracking Methods 1. FIB Culturing ▫ Total Coliforms, E. coli , Enterococcus 2. DNA marker identification by qPCR ▫ 2 human, bird, dog, horse 3. Other analyses depending on DNA results: ▫ Chemical analysis for caffeine & cotinine ▫ Community analysis (TRFLP) ▫ Viral analysis
Moving Forward � Determine if fecal contamination is present � Determine source(s) of contamination � Preliminary results are showing a couple areas where FIB may be originating: 1. Arroyo Burro Lagoon – Natural breaching of the lagoon may impact surf zone water quality 2. Las Positas Creek – Elevated FIB levels seen in sampling locations along this creek
Lagoon Closed
Lagoon Open
Lagoon Open
Camera on Lifeguard Tower
Thank You! � UCSB Holden Lab � Funding for the SIPP project comes from California ▫ Dr. Trish Holden Proposition 84 through the ▫ Laurie Van De Werfhorst Clean Beaches Initiative (CBI) ▫ Ning Jiang ▫ Ngoc Hoang ▫ Corinne Dorais � City Creeks Division ▫ Dr. Jill Murray ▫ James Rumbley � County Environmental Health ▫ Willie Brummett Questions/Comments? Contact: lagoon_study@bren.ucsb.edu
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