Microbial Source Tracking at Arroyo Burro Beach Jared Ervin Holden - - PowerPoint PPT Presentation

Microbial Source Tracking at Arroyo Burro Beach

Jared Ervin Holden Lab, UCSB

• Sept. 20th, 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 1st thru October 31st ▫ 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 FC/TC Ratio N/A N/A TC>1,000 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 Other tools needed to identify and locate sources

• f fecal pollution

FIB Culturing

FIB

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

Bird Specific Bacterial DNA Cow Specific Bacterial DNA Human Specific Bacterial DNA

How DNA-based Tools Work: qPCR of Human Bacteroides marker

• 1. Sample the water
• 2. Capture bacteria
• 3. Extract DNA
• 4. Amplify target

DNA (qPCR)

• 5. Quantify human

Bacteroides in water sample

1 copy 2 copies 4 copies 8 copies 20x = 1 million copies

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

▫ Dr. Trish Holden ▫ Laurie Van De Werfhorst ▫ 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 Funding for the SIPP project comes from California Proposition 84 through the Clean Beaches Initiative (CBI)