1. Overview of pathogens and indicator bacteria of Ambient conditions and monitoring for livestock associated pathogens and indicators in CA concern. waterways 2. Indicator bacteria dynamics in runoff from

E. coli rangeland and irrigated pasture.

3. Correlations between indicator bacteria and pathogens of concern.

C. hild C. parvum Salmonella

U.S. waterborne disease outbreaks causing gastroenteritis 1989 through 1996 Type of Agent No. Drinking Water organism outbreaks water recreation Protozoa Giardia duodenalis 27 18 9 Cryptosporidium 21 8 13 Bacteria E. coli O157:H7 11 3 8 Campylobacter jejuni 3 3 Salmonella 2 1 1

Pathogens and produce: rangeland runoff and irrigation water C. parvum E. coli Salmonella

A decade of produce outbreaks traced back to CA Livestock Pathogens of Waterborne & Year Food Vehicle Pathogen Cases Public Health Concern:

1996 Mesclun lettuce E. coli O157:H7 61 Protozoa: “hard” to eliminate during water treatment, 1996 Unpasteurized apple juice E. coli O157:H7 70 low infectious dose, mild to moderate illness

1996-1998 Alfalfa or clover sprouts E. coli O157:H7 600 ™ Cryptosporidium parvum (6 outbreaks) Salmonella ™ Giardia duodenalis 2000-2001 Raw almonds Salmonella 168 Bacteria: “easy” to eliminate during water treatment, 2002 Romaine lettuce E. coli O157:H7 29 2002-2004 Raw almonds Salmonella 47 higher infectious dose, mild to serious illness

2003 Baby spinach E. coli O157:H7 16 ™ pathogenic E. coli (e.g., Stx 1&2, O157:H7) 2006 Baby spinach E. coli O157:H7 205 ™ Salmonella 2006 Iceberg lettuce E. coli O157:H7 77 ™ Campylobacter 2006 Iceberg lettuce E. coli O157:H7 80

1 Indicator bacteria v. pathogens Ideal world: strong, reliable correlation between indicator bacteria and bovine pathogens in water total coliforms, fecal coliforms, indicator 100 E. coli, Enterococcus 75 Bacteria that when present in water indicate the 50 (oocysts / / L) (oocysts presence of fecal material and pathogens. / L) (oocysts

25 A standard is born C. parvum C. parvum 0 0 50 100 150 200 250 300 350 400 450 500

C. parvum E. Coli O157:H7 Salmonella indicator E. coli (cfu/100ml)

Indicator Bacteria Standards: Surface Waters Indicator Bacteria: One Big Happy Family

Fresh water standards exist for both “indicator” E. coli and fecal coliforms across CA: varies by water board total coliforms

USEPA Recommends E. coli fecal coliforms 1. geometric mean <126 bacteria per 100 ml from 5+ samples in 30 indicator days E. coli

2. single grab samples should not exceed 235 bacteria per 100 ml

E. coli: A malcontent in every family Beef cattle - indicator bacteria

indicator E. coli Fecal coliforms: outbreak pathogenic strain 10,000,000 to 100,000,000 per Shiga-toxin E. coli gm feces – all classes

Indicator E. coli: E. coli O157:H7 1,000,000 to 10,000,000 per gm

feces – all classes.

2 Indicator dynamics in runoff from rangeland and Small watershed scale – annual rangelands irrigated pasture.

Spatial Scale: Pasture to the watershed to the • Experimental watersheds Delta. HREC + + SFREC • HREC (7), SFREC (4) Temporal Scales: Runoff event to the water year. • Grazing and fire treatments Ramifications for monitoring and meeting water • 20 to 300 acres in size quality standards.

SFREC – heavy grazing/rangeland – can observe an order of SFREC – heavy grazing/rangeland – can observe an significant Schubert Watershed Winter Storm Dec. 12-17, 2001 reductionWatershed in 4: concentration 2002 Winter Stormflow from start to end of rainfall season. magnitude300 acre catchmentor greater on UC-SFREC change in Sierra Nevadaduring foothill aoak storm woodlands, event. grazed by beef cattle 300 acre catchment on in Sierra Nevada annual range, winter graze, cow-calf, 500 lb/ac RDM 9 25,000 4 10,000 Streamflow (cfs)

Streamflow (cfs) mL) coli (cfu/100 E. & Fecal Colif.

E.coli (cfu/100ml) Fecal Colif. & coli (cfu/100E. mL) 20,000 8,000 E.coli (cfu/100ml) 3 FC (cfu/100ml) 6 FC (cfu/100ml) 15,000 6,000

2 10,000

4,000 Streamflow (cfs) 3 Streamflow (cfs) Streamflow

1 5,000 2,000

0 0 0 0 3-Apr 9-Mar 2-Nov 0:00 0:00 0:00 0:00 0:00 0:00 28-Apr 17-Jan 11-Feb 27-Nov 22-Dec 12:00 12:00 12:00 12:00 12:00 12:00

Two storms in mid December One rainfall-runoff season

Valley and Foothill Irrigated Pastures HREC – observe variable differences between moderately grazedMean and generic non-grazed E. coli and watersheds, fecal coliform concentrations background from does 4 oak not = 0. woodland watersheds at UC-HREC near Hopland, CA during 2001 WY. Flood Irrigated Pasture Moderately grazed 3000 E. coli fecal coliform 2500

2000 Non-grazed since 1955

1500

1000

Concentration (cfu/100mL) Concentration 500 126 0 ABCD Watershed

3 E. coli Levels in Irrigated Pasture Tailwater E. coli Levels in Irrigated Pasture Tailwater A 7 hour irrigation event 8 Irrigation Events

1.6 80000 200000 Runoff tailwater 1.2 E. coli 60000 irrigation water 150000

0.8 40000 100000 (cfu/100ml) cfu/100ml grazing management E. coli

0.4 20000 E. coli 50000 Tailwater Rate (cfs) Runoff

0.0 0 0 07000900110013001500 Jun. Time of Day Sep.

Two Year Survey of WQ in 24 California rangeland streams Indicator E. coli and FC concentrations were dependent

Mean generic E. coli and fecal coliform concentration measured for 2 yearsupon on 24 rangeland watershed streams hydrologyacross California – (n=947 elevation samples) 2000 & 2001 + + 9,000 + Rainfall-RunoffRainfall-Runoff Snowmelt ~1000 grab samples under storm & Snowmelt and base flow conditions + + + + + 6,000 + + + E. coli + + 3,000 fecal coliform Concentration (cfu/100ml)Concentration

0 BFGHI KNQVXACEPTUDJLMORSW Stream

: Indicator E. coli and FC concentrations were variable Delta Project: Mean monthly indicator bacteria concentrations across 14 coastal and 88 sites monitored once a month for two years acrossfoothill annual the water rangeland year streams on annual WY 2000-2001. range watersheds. Bacterial indicators: 12000 fecal coliforms, E. coli, Enterococcus fecal coliforms 10000 E. coli Bacterial pathogens:

8000 mobilization of in-stream sediment Salmonella, shigatoxin 1&2 E. coli, Campylobacter

6000 irrigated pasture return +

cfu/100 mL environmental growth? 4000

2000

0 Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep

4 SAC Indicator E. coli 6/06-6/07, 12/07-11/08, n=1892 80% 70% of samples < 25 cfu 60%

RIO VISTA 40% Frequency

20% 5% of samples > 235 cfu

0% 0 50 100 150 200 250 STOCKTON STOCKTON E. coli (cfu / 100 ml)

Unknown correlations between indicators and indicator pathogens livestock associated pathogens for CA watersheds. bacteria = in water 100

Ideal Correlation ? 75

50 (oocysts / / L) (oocysts (oocysts / / L) (oocysts

25 C. parvum C. parvum 0 0 50 100 150 200 250 300 350 400 450 500 indicator E. coli (cfu/100ml)

Consider the shedding of pathogens and indicators in Annual Rangeland – SFREC & HREC 8 grazed rangeland watersheds – 2 years CA range and pasture cattle feces. Trace C. Parvum levels in <5% of ~600 stream samples

9 12,000 C. parvum in CA Indicator E. coli in CA range cattle range cattle Streamflow (cfs)

E.coli (cfu/100ml) coli E. 100 6 8,000

cows and calves (cfu/100 mL) calves 100% 5-15% cows 100,000 – 1,000,000 / g

100,000 / g (cfs) Streamflow <1% 3 4,000

% infected 1-100 / g 0 4812+ 4812+ 0 0 cattle age (months) 11/2/01 11/27/01 12/22/01 1/17/02 2/11/02 3/9/02 4/3/02 4/28/02

5 UFRW and Bridgeport Valley: 2007-08 Graze Season (May – Oct) 2007-08 Pathogen monitoring Sample monthly: indicator E. coli and FC, C. parvum, Salmonella, UFRW and Bridgeport Valley E. Coli O157:H7, Campylobacter 102-116 water samples taken, May-Oct Sample a total 16 sites: entering and exiting irrigated agriculture areas indicator E. coli < 235 cfu/100 ml > 235 cfu/100 ml Irrigation, beef cattle grazing, hay production Crypto 8=Yes 5 of 75 (6%) 3 of 27 (11%)

Salmonella 12=Yes 9 of 75 (12%) 3 of 27 (11%)

Campy 0=Yes 0 of 75 (0%) 0 of 27 (0%)

E. coli O157:H7 6=Yes 4 of 95 (4%) 2 of 21 (9%) 1,000 to 20,000 AU 1,500 to 32,000 ac irrigated

SAC 2007-08 Pathogen monitoring SAC UFRW and Bridgeport Valley

above meadow below meadow RIO Crypto 8=Yes 5 3 VISTA

Salmonella 12=Yes 10 2

Campy 0=Yes 0 0

O157:H7 6=Yes 0 6 STOCKTON STOCKTON

Delta pathogen – indicator monitoring Not so Ideal World: indicator E. coli concentrations not well correlated to Salmonella, C. parvum, etc. 955 water samples, 2006-07 1

indicator E. coli Mean Conc. < 235 cfu/100 ml > 235 cfu/100 ml

(cfu/100mL) 0.5 Indicator E. coli 22 cfu 470 cfu

Salmonella 0.30 MPN 0.25 MPN Salmonella 1,829 E. coli isolates from across these 955 samples 0 2 of 1,829 had Stx 1 (0.1%) 0 200 400 600 800 2 of 1,829 had Stx 2 (0.1%) indicator E. coli (cfu/100mL)

6 Possible Correlation at higher indicator levels? Summary 1000

• 800 Significant indicator bacteria associated with direct range and pasture runoff, 600 • Conc. reduction with increased spatial 400 scale: pasture>watershed>delta, 200 Pathogen (no. / 100 ml) 100 (no. / Pathogen Pathogen (no. / 100 ml) 100 (no. / Pathogen • Significant influence of watershed 0 hydrology and livestock management 0 2,000 4,000 6,000 8,000 10,000 on conc., Indicator bacteria (no. / 100 ml) Conc a ithin noff e ents (ho l )

Summary

• We consistently find very low levels of pathogens in these waters,

• Essentially non-existent correlations between pathogens and indicator bacteria in these waters,

• Management opportunities do exist to reduce microbial pollutant loads from these systems next topic

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