Toxic cyanobacterial risk to drinking water in the Willamette Valley
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1 Toxic cyanobacterial risk to drinking water in the Willamette Valley Connie S. Bozarth, Jon Shepardson, Nathan Brown, Theo Dreher Department of Microbiology and Soyoun Ahn, Jeff Morré, Claudia Maier Department of Chemistry Oregon State University, Corvallis, OR
2 Toxic cyanobacterial risk to drinking water in the Willamette Valley
3 Microscopic observation and colony isolation Pick colony, rinse 3x and divide: 25 µl for DNA analysis and 25 µl for toxin analysis
4 Sample preparation Samples are split: (a) Filtered for immediate use or storage in freezer for subsequent DNA extraction (b) (c) Frozen and thawed (to break cells) and stored frozen for toxin analysis Colonies are picked out under the microscope, frozen and thawed, and divided for DNA and MS (toxin) analysis (b) (a) (c) 0.45 micron glass fiber filter for DNA analysis Frozen & thawed water samples for toxin or DNA analysis Lysed colonies for for DNA analysis
5 The Samples Total no. of Samples Dexter Res. Lowell Raw Water Springfield SUB Dorena Res. Cott. Gr. Raw Water Detroit Res. Salem WTP June 28- Nov 30 July 28- Nov 30 State Park Covered bridge Hwy 58 Aug 15- Oct 5 Aug 25- Oct 3 Raw water Filter bed Sep 13- Oct 17 Bake Stewart Sep 13- Oct 11 June 28- Oct 26 Heater Ck Blowout Ck Log boom Aug 25 Filter bed Posted Bloom Aug 19 - Oct 14 Sep 19 - Oct Apr 12- Nov 11 Apr 12- Oct 16 Aug 14- Oct 16 Aug 15- Oct 17 Apr 12- Oct 17 July 25- Oct 17 May 9- Nov 11 June 6- Oct State Park Covered bridge Hwy 58 Mid-lake Raw water Filter bed Bake Stewart Baker Bay Heater Ck Blowout Ck Log boom Filter beds Posted Bloom Aug 13 Nov 16 July 31 Oct 23
6 Dexter Reservoir sampling sites Willamette River outflow Dexter Reservoir State Park Lowell Mid-lake Hwy 58 Covered bridge Raw water intake Upstream Lookout Point Reservoir Springfield WSSFF, 13 miles downstream WSSFF, SUB Willamette Slow Sand Filtration Facility
7 Dorena Reservoir sampling sites Dorena Reservoir Row River outflow Baker Bay Bake Stewart Cottage Grove water utility intake 4.6 miles downstream
8 Detroit Reservoir sampling sites Nth Santiam River outflow Log boom Detroit Reservoir Baker Bay Heater Ck Blowout Ck Bake Stewart Salem water utility intake (Geren Island) 30 miles downstream
9 What cyanobacteria are present? The reservoirs are dominated by Anabaena & Aphanizomenon Morpho logy Picture ID ID Size (µm) Dexter 2011 Dexter 2012 Dorena 2011 Dorena 2012 Detroit x DEXL Anabaena circinalis V: 8 H: 10 A: 12 x x DEXL Anabaena crassa V: 5 x 10 H: 10 A: x DEXL Anabaena lemmermannii V: 4 x 8 H: 6 x 10 A: 8 x 25 40x DORL Anabaena lemmermannii V: 6 x 8 H: 8 A: 8 x x DORL Anabaena planktonica V: 10 x 13 H: 11 A: x DEXL Anabaena flos-aquae V: 6 H: 6 x 8 A: x CR Microcystis aeruginosa V: 5 200x DORL Gloeotrichia echinulata V: 10 x 8 to 3 x 100; H: 8 A: x DEXL Aphanizomenon flos-aquae V: 5 x 5-15 H: 5 x 10 A: ---- Detroit 2012
10 What toxins are present? Analysis of water samples Toxin Assay type Year Dexter Res. Lowell Raw Water Springfield SUB Dorena Res. Cott. Gr. Raw Water Detroit Res. Salem WTP Microcystin Toxin detection mcye / 26 0 / 8 0 / 8 1 / 12 0 / 5 0 / 6 0 / / 19 0 / 5 0 / 10 3 / 28 0 / 13 1 / 13 0 / ± / 26 1± / 8 0 / 8 2± / 12 0 / 5 2 / 13 0 / / 25 0 / 5 2 / 10 1 / 28 0 / 13 4 / 10 1± / 5 Anatoxin-a Toxin detection anac / 6 0 / 3 0 / 6 0 / 3 0 / 2 0 / / 25 0 / 5 0 / 10 0 / 27 0 / 13 0 / 10 0 / / 26 1± / 8 2+2± / 9 0 / 10 0 / 5 1± / 13 0 / / 26 0 / 5 0 / 10 2 / 28 0 / 13 0 / 10 0 / 5 Cylindrospermopsin Toxin detection cyra, cyrj / 25 0 / 5 0 / 10 0 / 27 0 / 13 4 / 11 0 / / 13 0 / 8 0 / 9 5 / 13 0 / / 26 0 / 5 0 / 10 2 / 28 0 / 13 8 / ± / 5 detection only Toxin detection
11 Positive toxin or toxin gene hits: Dexter Date Location Microcystin µg/l MS ELISA MC Anatoxin Ana Cylindr osperm opsin Cyr Aug 19 State Park + Cov-Br nd mcye nd nd 2011 Aug 22 Lowell Raw water nd ± mcye nd ± anac Sep 5 Cov-Br nd ± mcye nd 2012 June 20 June 20 State Park Plankton net Cov-Br Plankton net nd nd nd nd nd cyra* cyrj* nd nd nd nd nd nd cyrj* Sep 7 SUB Raw water nd nd nd ± anac nd Sep 14 SUB Raw water nd nd nd anac nd 2011 Sep 20 SUB Raw water nd nd nd anac nd Sep 27 SUB Raw water nd nd nd ± anac nd Aug 20 SUB Raw water nd nd nd nd anac* nd nd 2012 Sep 21 SUB Raw water nd nd nd nd anac* nd nd *Verified by sequencing product; nd, not detected No measured toxin
12 Positive toxin or toxin gene hits: Dorena Date Location Microcystin (µg/l) MS ELISA MC Anatoxin Ana Sep 27 Bake Stewart YR nq nd nd Cylindr osperm opsin Cyr 2011 Oct 4 Bake Stewart nd ± mcye nd 2012 Oct 11 Bake Stewart nd ± mcye nd June 20 June 20 July 25 Bake Stewart Plankton net Baker Bay Plankton net Bake Stewart nd nd mcye* Nodularin *Verified by sequencing product; nd, not detected; ELISA, LOD = 0.05 µg/l; (Av. of 0.058, nd) Measured toxin near background levels and only at Bake Stewart nd anac* Oscillatoria nd nd nd nd nd nd ± LR Aug 22 Bake Stewart nd Sep 18 Bake Stewart nd nd ± ± nd anac* Oscillatoria nd nd cyra* cyrj* nd nd nd nd nd nd nd nd nd nd nd Sep 24 Baker Bay nd nd nd nd nd cyrj
13 Positive toxin or toxin gene hits: Detroit Date Location Microcystin µg/l MS ELISA MC Anatoxin *Verified by sequencing product; nd, not detected; (1) mcye sequence corresponding to Anabaena (all other mcye Microcystis-like) Ana Cylindr osperm opsin Cyr June 28 Heater, Hoover, Breitenbush & Blowout (each) nd nd cyrj 2011 Aug 16 Heater Ck nd mcye nd nd Aug 20 Heater Ck mcye nd nd nd Aug 24 Heater Ck nd nd nd ± anac nd Oct 26 Log Boom nd nd nd nd cyrj May 9 Heater Ck nd nd mcye* nd nd nd cyra cyrj* May 9 Blowout Ck nd nd nd nd nd cyra cyrj* May 29 Heater Ck nd mcye* nd nd ± cyra* cyrj* June 5 Heater Ck nd mcye* nd nd nd cyra cyrj* 2012 June 5 Blowout Ck nd nd nd June 5 Log Boom nd nd nd nd ± ± cyra cyrj cyra* cyrj* June 12 Log Boom nd nd nd nd ± cyra* cyrj* Aug 14 Heater Ck nd nd mcye* Microcystis nd nd nd nd Nov 6 Heater Ck nd nd nd cyrj
14 Summary: toxin analyses Toxins were only detected in water samples from Detroit Res. and at Bake Stewart in Dorena Res. (MC, Cyl, no ANA). All toxin levels were low. No toxins were detected in Dexter Reservoir. No anatoxin- a was detected in any of the reservoirs. Toxicity was below levels of concern in 2011/12. BUT, toxin genes for MC, ANA and Cyl are present in all lakes, indicakng the potenkal for toxin produckon. They are mostly present at a low level in a minority of samples, except for the cyr genes in Detroit.
15 Colony genotype analysis at the cpcba gene locus reveals three main Anabaena/ Aphanizomenon genotypes Aphan. flos-aquae Anabaena Anabaena Relationship between genotype and morphology is uncertain
16 Deep sequencing analysis at the cpcba gene confirms the prevalence of three genotypes Aphan. flos-aquae c78 Anabaena c169 Anabaena c106 August September Aug Sep September October August September
17 Deep sequencing analysis at the cpcba gene confirms the prevalence of three genotypes Aphan. flos-aquae c78 Anabaena c169 Anabaena c106 August September Aug Sep All 3 genotypes are currently non-toxic. Which toxin genes do they carry? September October August September
18 Different cpcba genotypes are present in Detroit Reservoir genotypes: source of cylindrospermopsin? Aphan. flos-aquae c78 Anabaena c169 Anabaena c106
19 What have we learned? The blooms present in Dexter, Dorena and Detroit Res. are Anabaena/ Aphanizomenon- dominated, with several morphotypes and 3 major cpcba genotypes (c169, c106, c78) across the system. Some addikonal genotypes are present in Detroit Res. Toxins were only detected in water samples from Detroit Res. and at Bake Stewart in Dorena Res. (MC, Cyl, no ANA). All toxin levels were low. No toxins were detected in Dexter Reservoir. Toxicity was below levels of concern in 2011/12. But, toxin genes for MC, ANA and Cyl are present in all lakes, indicakng the potenkal for toxin produckon.
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MEDFORD/JACKSON CO. OR Latitude = 42.37 N WMO No. 72597 Longitude =122.8 W Elevation = 1329 feet Period of Record = 1973 to 1996 Average Pressure = 28.61 inches Hg Design Criteria Data Mean Coincident
Design Criteria Data
MIDLAND TX Latitude = 31.95 N WMO No. 72265 Longitude =2. W Elevation = 2861 feet Period of Record = 1973 to 1996 Average Pressure = 27.3 inches Hg Design Criteria Data Mean Coincident (Average) Values
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Design Criteria Data
ALBUQUERQUE NM Latitude = 35.5 N WMO No. 72365 Longitude =16.6 W Elevation = 5315 feet Period of Record = 1967 to 1996 Average Pressure = 24.72 inches Hg Design Criteria Data Mean Coincident (Average)
Design Criteria Data
DULUTH MN Latitude = 46.83 N WMO No. 72745 Longitude = 92.18 W Elevation = 1417 feet Period of Record = 1973 to 1996 Average Pressure = 28.43 inches Hg Design Criteria Data Mean Coincident (Average) Values
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LAS VEGAS/MCCARRAN NV Latitude = 36.8 N WMO No. 72386 Longitude =115.1 W Elevation = 2179 feet Period of Record = 1973 to 1996 Average Pressure = 27.67 inches Hg Design Criteria Data Mean Coincident (Average)
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PHOENIX/SKY HARBOR AZ Latitude = 33.43 N WMO No. 72278 Longitude =112. W Elevation = 116 feet Period of Record = 1973 to 1996 Average Pressure = 28.72 inches Hg Design Criteria Data Mean Coincident (Average)
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