samples · web viewnitrate-nitrite (dissolved inorganic nitrogen) was measured by cadmium reduction...
TRANSCRIPT
Supplement for
Proliferation of Microalgae and Enterococci in the Lake Okeechobee, St. Lucie, and
Loxahatchee Watersheds
E Kelly1,2,3, M Gidley3,5,6, C Sinigalliano3,5, N Kumar7, L Brand3,4, RJ Harris8, HM Solo-Gabriele1,2,3
1University of Miami Leonard and Jayne Abess Center for Ecosystem Science and Policy, Coral Gables, FL, USA
2University of Miami Department of Civil, Architectural and Environmental Engineering, Coral Gables, FL, USA
3NSF NIEHS Oceans and Human Health Center, Rosenstiel School of Marine and Atmospheric Science, University of Miami,
Miami, FL, USA
4University of Miami Department of Marine Biology and Ecology, Rosenstiel School of Marine and Atmospheric Science
(RSMAS), Miami, FL, USA,
5National Oceanic and Atmospheric Administration (NOAA) Atlantic Oceanographic and Meteorological Laboratory (AOML)
Environmental Microbiology, Miami, USA
6University of Miami Cooperative Institute for Marine and Atmospheric Studies (CIMAS), Miami, USA
7University of Miami Department of Public Health Sciences, Division of Environment & Public Health, Miami, FL, USA
8Loxahatchee River District, Jupiter, FL, USA
For consideration for potential publication in: Water Research
Version Dated: December 20, 2019
Corresponding author: Helena Solo-Gabriele, Ph.D.e-mail address: [email protected]
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Supplementary Table S1. GPS coordinates for sites and water control structures
Site Latitude (decimal degrees) Longitude (decimal degrees)Canal Point 26.866056 -80.631639S308 26.984892 -80.621428S80 27.111633 -80.284981Roosevelt Bridge 27.202444 -80.258SE03 27.202778 -80.259167Dubois Park 26.943389 -80.074556
3536
3738
Site Visit Sample Location Latitude Longitude Microcystin(µ/L) FDEP Algal ID8/27/2018 Leighton 27.17 -80.26 2.9 Dominant taxon: Microcystis aeruginosa8/27/2018 A1A bridge 27.20 -80.21 6.7 Dominant taxon: Microcystis aeruginosa8/27/2018 Roosevelt 27.20 -80.26 27.0 Dominant taxon: Microcystis aeruginosa8/27/2018 North Fork 27.21 -80.28 4.8 Dominant taxon: Microcystis aeruginosa8/23/2018 SLE South Fork 27.20 -80.26 14.0 Dominant taxon: Microcystis aeruginosa8/23/2018 SLE N Fork 27.22 -80.28 2.9 Dominant taxon: Microcystis aeruginosa8/23/2018 C44S80 St. Lucie lock 27.11 -80.28 495.1 Dominant taxon: Microcystis aeruginosa8/22/2018 Bathtub Beach 27.19 -80.16 1.2 Dominant taxon: Microcystis sp.8/20/2018 SF near Rustic 27.13 -80.26 16.0 Dominant taxon: Microcystis aeruginosa8/16/2018 SF near Leighton 27.17 -80.26 2.3 Dominant taxon: Microcystis aeruginosa8/14/2018 Lake Okeechobee L004 26.98 -80.71 11.0 Dominant taxon: Microcystis aeruginosa8/14/2018 Lake Okeechobee 27.05 -80.75 2.6 Dominant taxon: Microcystis aeruginosa8/13/2018 S308C Port Mayaca 26.98 -80.62 1.1 Dominant taxon: Microcystis aeruginosa8/9/2018 C44S80 St. Lucie lock 27.11 -80.29 1.0 Dominant taxon: Microcystis aeruginosa8/6/2018 SF near Meridian 27.18 -80.27 4.5 Dominant taxon: Microcystis aeruginosa8/6/2018 NF near Seagate 27.20 -80.27 10.0 Dominant taxon: Microcystis aeruginosa8/2/2018 St. Lucie Canal @ Martin Highway 27.16 -80.25 30.0 Dominant taxon: Microcystis aeruginosa8/2/2018 SF Canal 27.18 -80.26 5.9 mixed algae; no dominant species in sample8/2/2018 SE near Central Marine 27.22 -80.26 110.0 Dominant taxon: Microcystis aeruginosa8/2/2018 SE Warner Creek 27.22 -80.23 3.1 Dominant taxon: Microcystis sp.7/30/2018 S Fork 27.18 -80.27 1.3 mixed algae; no dominant species in sample7/30/2018 St. Lucie Canal 27.14 -80.26 8.6 mixed algae; no dominant species in sample7/26/2018 Outside Frazier 27.20 -80.26 3.2 Dominant taxon: Microcystis aeruginosa7/26/2018 Warner Creek 27.22 -80.23 2.1 Dominant taxon: Microcystis aeruginosa7/26/2018 Roosevelt 27.20 -80.26 1.2 Dominant taxon: Microcystis aeruginosa7/26/2018 C44S80 St. Lucie Locks 27.11 -80.28 3.1 mixed algae; no dominant species in sample7/23/2018 SLR Canal @ de la Bahia 27.17 -80.25 2.5 mixed algae; no dominant species in sample7/19/2018 SLE near Central Marine 27.22 -80.25 8.3 Dominant taxon: Microcystis aeruginosa7/19/2018 St. Lucie Locks, C44S80 27.11 -80.29 4.7 Dominant taxon: Microcystis aeruginosa7/19/2018 Mouth of Warner Creek 27.22 -80.23 6.9 Dominant taxon: Microcystis aeruginosa7/18/2018 LZ30 - Lake Okeechobee 26.80 -80.86 8.8 Dominant taxon: Microcystis aeruginosa7/17/2018 CLV10A inside Lake Okeechobee 26.92 -80.63 2.4 Dominant taxon: Microcystis aeruginosa7/17/2018 L004 26.98 -80.71 3.1 Dominant taxon: Microcystis aeruginosa7/16/2018 Circle Bay Condos 27.18 -80.26 2.8 Dominant taxon: Microcystis aeruginosa7/16/2018 Middle Estuary North Shore 27.21 -80.25 6.1 Dominant taxon: Microcystis aeruginosa7/16/2018 South Fork @ Rustic Circle 27.13 -80.26 3.0 Dominant taxon: Microcystis aeruginosa7/16/2018 SLE near Leighton 27.17 -80.26 5.0 Dominant taxon: Microcystis aeruginosa7/16/2018 S308C 26.98 -80.62 2.8 Dominant taxon: Microcystis aeruginosa7/12/2018 C44S80 27.11 -80.28 15.0 Dominant taxon: Microcystis aeruginosa7/5/2018 Lake Okeechobee NW of Port Mayaca 26.99 -80.67 4.0 mixed algae; no dominant species in sample7/5/2018 Lake Okeechobee SW of Port Mayaca 26.94 -80.65 6.0 mixed algae; no dominant species in sample7/5/2018 C44S80 St. Lucie Lock 27.11 -80.28 154.4 Dominant taxon: Microcystis aeruginosa7/2/2018 SLE near Central Marine 27.22 -80.25 1.9 Dominant taxon: Microcystis aeruginosa7/2/2018 St. Lucie Estuary Poppleton Creek 27.19 -80.26 1.1 Dominant taxon: Microcystis aeruginosa7/2/2018 Lake Okeechobee S308C lakeside 26.98 -80.62 1.9 Dominant taxon: Microcystis aeruginosa6/27/2018 St. Lucie Locks, C44S80 27.11 -80.29 1.3 Dominant taxon: Microcystis aeruginosa6/25/2018 Port Mayaca 26.98 -80.62 2.2 Dominant taxon: Microcystis aeruginosa6/18/2018 Lake_Okeechobee_S352 26.86 -80.63 5.9 Dominant taxon: Microcystis aeruginosa
Supplementary Table S2. Results of surface water samples collected from Palm Beach and Martin Counties during the 2018 cyanobacteria bloom. Table displays all results over the established laboratory detection limits. Samples were collected by FDEP or SFWMD staff and samples analyzed at FDEP laboratory. Data access https://floridadep.gov/dear/algal-bloom/content/algal-bloom-sampling-results
39404142434445
SUMMARY OF ANALYTICAL METHODS
1. Chlorophyll a and pheophytin
In this study, both spectrophotometry (e.g., monthly field sampling) and the fluorometric
method (e.g., microcosm experiments) were used to measure chlorophyll a and pheophytin. The
measurement of chlorophyll a is widely used to provide a measure of microalgae biomass during
blooms; confirmatory tests were performed by the Florida Department of Environmental
Protection (FDEP) to determine the presence of cyanobacteria. Chlorophyll a values are
generally reported as their corrected value, which is the value after acidification and are reported
as “chlorophyll a” in μg/L herein. Pheophytin is also reported in μg/L.
The spectrophotometry method was based on the Standard Methods 22nd Edition (APHA
2012), as modified by the Loxahatchee River District (LRD). Samples were filtered on Whatman
0.47 μm glass microfiber filters and preserved with 1 mL of 1% saturated magnesium carbonate
solution within 24 hours of collection and stored frozen. Within 28 days of collection samples
were ground (1 minute tissue grinder) and set in 90% aqueous acetone for approximately 24
hours; the mixture was centrifuged at 4000 rpm for 20 min to separate the ground filter from the
supernatant. The supernatant was analyzed spectrophotometrically (Perkin-Elmer Lambda 365
UV/Visible spectrophotometer), in which the sample was exposed to six different wavelengths of
light, within a range of 480-750 nm. The absorption was recorded for each wavelength. The
samples were acidified and re-exposed to the same six wavelengths of light; the absorbance was
recorded. The lowest practical range of determination with the spectrophotometric method used
was 1.0 µg/L. All samples were processed under dark conditions.
In the fluorometric method, 100 mL (or until the filter clogged) of sample water was
filtered on Whatman 47 mm glass microfiber filters. The filters were placed in vials and frozen,
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to be analyzed within 28 days. For analysis, the filters were transferred to a glass tube and 10 mL
dimethyl sulfoxide (DMSO) was added. The filter and DMSO set for 30 minutes under dark
conditions. Fifteen mL of acetone was then added, agitated, and the mixture was stored in the
freezer for 24 hours. Fluorometry (Turner Designs 10-AU Fluorometer with infrared sensitive
photomultiplier tube, calibrated with pure chlorophyll a) was then used to analyze the mixture
before acidification and after (to produce the degradation product pheophytin). The range can be
adjusted through dilution. All chlorophyll samples were processed under dark room conditions.
2. Enterococci
Enterococci was typically measured (n=11) using the membrane filtration (MF) method
(EPA method 1600; EPA 2009). One exception was made where the sample was analyzed using
chromogenic substrate (APHA 2012). For membrane filtration, 10 mL of sampled water was
filtered through a 0.45 μm sterile glass fiber membrane. The membrane was placed on selective
media (mEI) and incubated at 41ºC for 24 hours. The number of colony forming units (CFU)
present on the membrane were counted and multiplied by a dilution factor of 10 to provide units
of CFU/100 mL (EPA 2009). For chromogenic substrate, one pack of Enterolert chromogenic
media (IDEXX) was added to 100 mL of sample (saline sites were diluted 1:10 as stated in the
IDEXX protocol) and shaken to dissolve all chromogenic media. The mixture was then added to
a 51-Well Quanti-Tray, sealed, and incubated at 41ºC for 24 hours. Results were read under an
ultraviolet light and the number of wells that fluoresced were counted as positive. The number of
fluorescent wells were used to determine the Most Probable Number (MPN) from the MPN table
provided by IDEXX based on the 95% confidence interval; this MPN was multiplied by the
dilution factor to provide units of MPN/100 mL.
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3. Nitrate-Nitrite
Nitrate-Nitrite (dissolved inorganic nitrogen) was measured by cadmium reduction (EPA
353.2 and APHA 2012) using a flow injection analyzer (FIA) with autosampler (Lachat
QuikChem 8500 series) on 0.45 μm field-filtered samples. Samples were processed within 48
hours of collection. An ammonium chloride buffer pH adjusted to 8.5 with EDTA added to
reduce metal interference and sulfanilamide color reagent (10% concentrated phosphoric acid,
sulfanilamide and N-(1-naphthyl) ethylenediamine dihydrochloride) were injected as sample
flowed through a copper-treated cadmium column. Nitrate-nitrite was determined through the
intensity (absorbance) of the pink color produced by the sample according to the concentration;
calculated by a known standard calibration curve (R2 ≥ 0.999; n=6). Samples were read via
spectrophotometer flow cell with an absorbance wavelength of 520nm. Nitrate-nitrite was
measured in units of mg/L. The practical range of the determination was from 0.005 mg/L to 0.3
mg/L.
4. Total Kjeldahl Nitrogen (TKN)
The determination of total Kjeldahl nitrogen (TKN) is based upon the conversion of
organic sources of nitrogen (free ammonia, ammonia nitrogen, amino acids, proteins, and
peptides) to ammonium sulfate (EPA Method 351.2 1993 and APHA 2012). Samples were
preserved to pH < 2 with sulfuric acid and processed within 28 days. Samples were boiled at 200
°C for 2 hours to reduce organic nitrogenous components. Samples were then digested at 380 °C
for one hour in a solution of potassium sulfate, sulfuric acid, and cupric sulfate (copper) as the
catalyst. Measurements were then made using FIA with autosampler (Lachat QuikChem 8500
series) spectrophotometer (absorbance of 660nm), and standard calibration curve methods
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mentioned above. TKN is measured in units of mg/L. The practical range for the determination
was from 0.2 mg/L to 5 mg/L.
5. Total Nitrogen
Total nitrogen in this paper was reported as “nitrogen,” in mg/L. It was not measured
directly but was calculated by adding nitrate-nitrite and TKN.
6. Phosphorus and orthophosphate
Samples were analyzed for total phosphorus by persulfate oxidation digestion (121°C for
30 minutes). This process oxidizes all available forms of phosphate to orthophosphorus, thus
providing a measure of total phosphorus (TP). TP was then determined colorimetrically by
molybdate/ascorbic acid colorizing reagent (APHA 2012 4500-P A and standard method 4500-P
E) and analyzed spectrophotometrically (Perkin-Elmer Lambda 365 UV/Visible
spectrophotometer), with a practical range of 0.005 mg/L to 0.50 mg/L. Samples analyzed for
orthophosphate (dissolved reactive phosphorous) were field-filtered through a 0.45 μm
membrane filter and processed within 48 hours by colorimetric analysis without persulfate
oxidation digestion (SM4500-P F). Ammonium molybdate and antimony potassium tartrate
were combined as a color reagent and a 0.33N ascorbic acid buffer was added to create a blue
color change proportionate to orthophosphate concentration. Measurements were made using
FIA with autosampler (Lachat QuikChem 8500 series), spectrophotometer (absorbance of
880nm), and standard calibration curve methods mentioned above. The practical range of
orthophosphate was from 0.005 mg/l to 0.50 mg/l.
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7. Turbidity
Turbidity was measured in the laboratory within 48 hours using a Hach 2100Q turbidity
meter calibrated using 3 formalin standards (APHA 2012 method D1889-88A and EPA 180.1).
Results were reported in nephelometric turbidity units (NTU). The range of analysis was 0.1-
4000 NTU.
ADDITIONAL DETAILS ON LABORATORY QUALITY CONTROL ANALYSES
1. Quality Control for Nitrogen, Phosphorus, Chlorophyll, turbidity, conductivity, turbidity,
dissolved oxygen, and enterococcus by chromogenic substrate (EnterolertTM).
These parameters were analyzed through the Loxahatchee River District (LRD), a
National Environmental Laboratory Accreditation Program (NELAP) certified
laboratory. This laboratory uses EPA’s standard methods and also Standard Methods for
the Examination of Water and Wastewater, 22nd edition. The specific method of each
analyte and quality control criteria are given in Table S2.
The EPA methods and standard methods both state that the determined concentrations
from each method must be within +/-10% of the control sample (i.e., secondary standard
or the quality control sample). The analytical runs used for all of our samples fell within
this criterion.
The exception is chlorophyll a. As there is no published QC criteria for chlorophyll a,
historical data was used to determine QC acceptance criteria (+/-60%, based on the ability
to detect the true value of the control samples).
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Table S3: Table of Quality Control Acceptance Criteria for Loxahatchee River District Laboratory Activities
Parameter MethodBlank(mg/L)
LOQ (mg/L)
# Cal or Initial Stds.
CalibrationR2 or
%recovery2nd Std.
% recovery
Continuing Cal. Std. %
recovery LCS
Dup. Precision
RPD
Spike Accuracy
% recovery Hold TimeMethod of
Preservation
Enterococcus
Enterolert MPN
1 per lot of Quanti-Tray
10 MPN/100
ml
3 control organisms per lot of Enterolert
Within mfg accept criteria
N/A N/A(4) <51 &
Rlog < 0.361N/A 8 hrs
On ice,Refrigerate
< 6°C
TKNBlock dig.
FIAEPA 351.2
1 pre, (4)
1 post.< LOQ
0.2 6 to bracket sample range ≤ 0.995 90 – 110(1) 90 – 110(1) < 20(4) 90 – 110(3,4) 28 d H2SO4
pH < 2
NO2 + NO2Cd reductionFIA (low)
EPA 353.21 pre, (4)
1 post.< LOQ
0.005 6 to bracket sample range ≤ 0.995 90 – 110(1) 90 – 110(1) < 20(4) 90 – 110(3,4) 48 hrs No pres.
Field filtered
NO2 + NO2Cd reductionFIA (high)
EPA 353.21 pre, (4)
1 post.< LOQ
0.02 6 to bracket sample range ≤ 0.995 90 – 110(1) 90 – 110(1) < 20(4) 90 – 110(3,4) 28 d H2SO4
pH < 2
Ortho-Phoscolorimetric
SM4500-P E1 pre, (4)
1 post.< LOQ
0.005 6 to bracket sample range ≤ 0.995 90 – 110(1) 80 – 120(1) < 20(4) 90 – 110(3,4) 48 hrs No pres.
Field filtered
Total Phosphoruscolorimetric
SM4500-P E1 pre, (4)
1 post.< LOQ
0.005 6 to bracket sample range ≤ 0.995 90 – 110(1) 80 – 120(1) < 20(4) 85 – 115
(3,4) 28 d H2SO4pH < 2
Chlorophyll aSpectrophotometer SM10200H
1 pre, (4)
1 post.< LOQ
1 µg/L 1 in duplicate 40 - 160 N/A 40 - 160 < 30 N/A 28 dGlass filter, freeze
Conductivity,Electronic Meter EPA 120.1 1 pre, (4)
< LOQ2
µmhos/cmMinimum 2,Bracket range 95 – 105 95 – 105(1) 95 - 105(2,4)
+ at end ≤ 5 (4) N/A 48 hrsOn ice,Refrigerate
< 6°C
Turbidityturbidimeter EPA 180.1 1 DI H20(9)
< LOQ0.1 NTU 3 formazin,
quarterly 90 – 110
90 - 110(1)
1 10 NTU Formazin
std.
90 – 1101 10 NTU Formizin every
10 samp + end
≤ 10 (4) N/A 48 hrsOn ice,Refrigerate< 6°C
pHelectronic meter EPA 150.1 N/A N/A 2 – 3,
bracket range
95 – 105% eff.
of probe
±0.2 pH units ±0.2 pH units ≤ 5 N/A Within 15
min None
Dissolved Oxygenelectronic EPA 360.1 N/A N/A
1 initial, water saturated air
±0.5 mg/LConsult chart N/A
95 – 1051 at end,Saturated air
N/A N/A Immediately None
Footnotes for Table of Acceptance Criteria1) Secondary standards verify validity of method or calibration performance; Run after calibration or initial standards but before samples. 2) Laboratory Control Standards are fortified blanks that verify continuing method performance and are prepared in duplicate of the same analyte concentration as the spike. Blank used to prepare LCS must be treated same as sample; i.e. acid preserved, pH adjusted, filtered, etc.3) Matrix spike is typically, but not always the same sample as matrix duplicate. Spike concentration should be less than half of calibration or analytical range. 4) Run every 10 samples or matrix batch.5) Must meet 2.0 mg/L minimum DO depletion (initial – final) and 1.0 mg/L residual (final) DO for each test bottle.6) Dried to a constant weight @ 180C; i.e. weight change < 0.0005g.7) Choose sample volume to yield < 200 mg residue.8) Choose sample volume to yield between 2.5 – 200 mg residue and complete filtration within 10 minutes.9) Run every 20 samples or matrix batch.
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2. Quality Control for Enterococci by Membrane Filtration
The reproducibility of enterococci analysis by membrane filtration (EPA method 1600) was
determined by collecting 6 samples and analyzing them in quadruplicate. For each sample,
the mean, standard deviation, and coefficient of variation (COV) (standard deviation divided
by the mean) was determined. The average value of the COV was determined as 81.2%.
This value was then used to determine the expected standard deviation of a particular sample
by multiplying the enterococci value for the sample by the COV of 81.2%.
3. Quality Control for Chlorophyll a and Pheophytin by Fluorometry.
The reproducibility of chlorophyll a and the pheophytin measurements by fluorometry (used
for microcosm experiments only) was determined through the analysis of 6 samples analyzed
in quintuplicate as documented by Brand (2002). Computations were completed in a similar
fashion as for enterococci by membrane filtration. The average value of the COV was
determined to be 83.7%. This value was then used to determine the expected standard
devation of a particular sample by multiplying the chlorophyll a value for the sample by the
COV of 83.7%.
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Sample D_10 D_30 D_60
Uniformity Coefficient
Coefficient of Gradation
Canal Point (incubator) 2.90 4.83 7.35 2.53 1.09Canal Point (algae room) 2.98 5.17 8.19 2.75 1.10Canal Point (microcosm 4) 4.90 6.78 9.77 1.99 0.96Dubois Park (microcosm 4) 0.18 0.28 0.41 2.24 1.00
Supplementary Table S4. Grain size analysis. All of the samples were poorly/uniformly graded, which means that they were distributed within a narrow size range. The three samples from Canal Point can be classified as gravels and the Dubois Park sample can be classified as sand, based on the Unified Soil Classification System (USCS).
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Variable Roosevelt Bridge R2
Dubois Park R2
Rainfall 24 Hrs Before
0.001 0.002
Rainfall 3 Days Before
0.04 0.002
Rainfall Week Before
0.04 0.001
Air Temperature
0.001 NS
Water Temp 0.03 NSSupplementary Table S5. Result of linear regression for temperature and rainfall (quantitative variables) versus enterococci levels at Roosevelt Bridge and Dubois Park, 2000-2018. NS corresponds to correlations that were not significant.
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Parameter Canal Point Roosevelt Bridge Dubois ParkChlorophyll (μg/L) 33.27 9.44 3.06
Pheophytin (μg/L) 28.23 11.85 2.62
ENT (CFU/100 mL) 526 232 84
Nitrate-Nitrite mg/L 0.41 0.19 0.02
TKN mg/L 2.15 1.05 0.36
TN mg/L 2.55 1.21 0.37
Orthophosphorus mg/L 0.10 0.11 0.01
Total Phosphorus mg/L 0.22 0.19 0.03
Sampling Depth (m) 0.24 0.51 1.00
Water Temperature (ºC) 26.29 24.76 25.03
pH 8.54 7.79 7.88
Salinity ppt 0.18 10.24 33.77
DO (mg/L) 7.00 6.46 4.84
Turbidity (ntu) 171.4 31.5 6.4
Air Temperature (F) 78.58 78.17 73.92
Wind (mph) 6.5 7.6 8.3
Humidity (%) 61.6 68.8 76.2
Supplementary Table S6. Monthly sampling means for chlorophyll, enterococci, nutrients, and basic physical-chemical and meteorological data.
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Site Month
Enterococci per 100 mL (CFU) or MPN*
Nitrite-
Nitrate
(mg/L)
TKN (mg-N/L)
TN (mg/L)
Orthophosphorus (mg/L)
Phosphorus (mg/L)
Chlorophyll a ug/L
(corrected)
Pheophytin (ug/L)
Canal Point
September 18 0.39 1.70 2.09 not analyzed 0.209 121.7 5.8
October 60 0.46 1.29 1.75 not analyzed 0.168 8.0 5.6November 336 0.47 2.82 3.29 not analyzed 0.260 17.4 22.5
December 104* 0.70 unavailab
le unavailab
le 0.072 0.254 11.2 11.2
January 30/<10* 0.69 2.60 3.29 not analyzed 0.170 9.6 21.5February 400 0.60 1.70 2.30 0.091 0.277 7.6 11.2March 1000 0.58 4.80 5.38 0.423 0.423 17.3 26.3April 10 0.44 2.30 2.74 0.239 0.239 46.9 51.9May 1650 0.19 1.40 1.59 0.121 0.121 6.3 10.7
June 1390 0.28 2.20 2.48 0.103 0.103 99.7 105.2July 1110 0.06 2.00 2.06 0.100 0.367 35.1 45.5August 200 0.03 3.00 3.65 0.075 0.08 18.4 21.4
Roosevelt Bridge
September 40 0.54 1.40 1.94 not analyzed 0.257 3.6 6.4
October 100 0.35 1.56 1.91 not analyzed 0.249 6.4 5.6November 112 0.39 1.68 2.07 not analyzed 0.192 4.5 4.5
December 97* 0.41 unavailab
le unavailab
le 0.101 0.168 4.4 2.8
January 20 /10* ≤0.02 0.70 0.70 not analyzed 0.118 21.4 31.4February ≤10 ≤0.02 0.50 0.50 0.064 0.089 4.8 6.7March 30 0.01 0.70 0.71 0.095 0.130 6.9 8.3April 10 0.02 0.40 0.42 0.103 0.149 4.1 5.7May 100 0.03 0.40 0.43 0.128 0.138 3.9 5.0June 1020 0.32 1.50 1.82 0.173 0.295 15.2 18.8July 860 0.08 1.50 1.58 0.127 0.253 21 24.0August 390 0.08 1.20 1.28 0.122 0.220 17.1 23.0
Dubois Park
September 5 0.01 0.50 0.51 not analyzed 0.034 4.5 2.4
October 10 0.04 0.55 0.59 not analyzed 0.047 10.1 3.0November 23 0.05 0.71 0.76 not analyzed 0.044 3 1.8
December 52* 0.03 unavailab
le unavailab
le 0.023 0.033 2.7 1.4
January 10 /≤10* 0.03 0.40 0.43 not analyzed 0.038 3 5.3February 40 0.02 ≤0.2 ≤0.2 0.009 0.015 2.2 2.8March 200 ≤0.005 0.50 0.50 ≤0.005 0.014 1.1 1.4April 30 0.01 ≤0.2 ≤0.2 ≤0.005 0.021 2 2.7May 330 0.01 ≤0.2 ≤0.2 ≤0.005 0.007 2.6 3.4June 110 0.01 0.30 0.31 0.013 0.015 1.7 2.1July 170 0.01 0.50 0.51 0.018 0.023 2.5 3.2August 30 0.03 ≤ 0.2 ≤ 0.234 ≤ 0.005 0.010 1.3 1.9
*=Most Probable Number (MPN).
Supplementary Table S7. Monthly sampling data for cyanobacteria, enterococci, and nutrients.
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230231
232233
Site monthTime of
collectionSample
Depth (m)Temperature
(˚C) pHTide or wave
height
Salinity (ppt) Dissolved Oxygen
(mg/L)
Turbidity from lab analysis
(NTU)
Canal Point
September 4:58 PM 0.60 34.6 9.2 >1 foot 0.22 unavailable 49.8 October 5:39 PM 0.32 31.1 8.0 >1 foot 0.19 5.29 62.8November 2:20 PM 0.30 24.5 8.4 2-3 feet 0.18 6.50 304December 12:55 PM 0.26 18.8 8.2 1-2 feet 0.18 8.61 516January 12:42 PM 0.15 13.2 8.1 1 foot 0.13 9.02 254February 12:48 PM 0.21 19.6 8.8 >1 foot 0.17 8.10 149March 1:14 PM 0.17 20.8 8.6 1-2 feet 0.18 8.73 384April 1:06 PM 0.06 30.0 8.9 >1 foot 0.18 8.93 70.5May 12:00 noon 0.09 27.6 8.3 >1 foot 0.19 8.02 70.7June 12:20 PM 0.26 31.7 8.6 1 foot 0.17 7.60 74.4July 12:39 PM 0.20 30.5 8.6 1-2 feet 0.18 6.78 81.4August 11:51 AM 0.21 33.1 8.8 1-2 feet 0.16 6.41 40.7
Roosevelt Bridge
September 2:04 PM 0.51 28.8 7.6 high 2.28 unavailable 47.2 October 4:01 PM 0.58 28.2 7.8 Flood tide 0.20 5.44 91.3November 12:26 PM 0.82 24.2 7.9 Flood tide 0.18 6.00 71.9December 11:02 AM 0.57 18.6 7.7 Low 2.20 7.46 28.7January 10:58 AM 0.36 15.5 8.2 Flood tide 12.40 10.54 15February 11:04 AM 0.34 21.6 8.4 Flood tide 17.55 8.23 8.52March 11:31 AM 0.57 19.7 7.8 Ebb tide 26.46 7.10 9.2April 11:14 AM 0.38 26.8 7.7 Ebb tide 27.29 5.33 7.2May 10:13 AM 0.59 25.4 7.3 Ebb tide 26.26 5.07 7.2June 10:25 AM 0.56 28.1 7.5 Ebb tide 1.89 5.30 50.7July 10:50 AM 0.57 30.4 7.8 Ebb tide 3.26 5.42 18.4August 10:09 AM 0.30 29.8 7.8 Low 2.87 5.22 22.1
Dubois Park
September 12:15 PM 0.51 29.4 8.0 high 35.10 unavailable 14.5 October 2:31 PM 0.97 29.6 7.8 Flood tide 24.30 4.90 4.59November 10:49 AM 1.10 25.0 7.8 Ebb tide 27.60 4.80 8.84December 9:31 AM 0.91 19.8 7.6 Flood tide 32.90 5.49 5.9January 9:22 AM 0.93 16.9 7.8 Flood tide 26.90 7.50 22.4February 9:29 AM 1.03 22.4 8.4 Low 36.86 6.45 4.83March 9:45 AM 1.27 21.6 7.9 Ebb tide 38.32 5.72 4.2April 9:35 AM 0.79 26.1 7.9 Ebb tide 37.65 4.86 3.9May 8:50 AM 1.19 25.7 7.5 high 37.90 5.16 1.8June 8:50 AM 1.38 26.9 7.9 Ebb tide 35.54 4.50 1.7July 9:08 AM 1.14 29.5 7.9 Ebb tide 34.47 4.01 3.6August 8:49 AM 0.8 27.4 8.0 Flood tide 37.67 4.73 0.6
Supplementary Table S8. Monthly sampling means for basic physical-chemical data
234235236237238239240241242243244
245246247248
Supplementary Figure S1. Microcosm experiment setups. CP=Canal Point, RB=Roosevelt Bridge, DP=Dubois Park.
unfiltered
filteredDPRBB
CP
Water only Algae room conditions 19ºC with
alternating light and dark cycles
DPRBB
CP
Microcosm Experiment 1
CP/DI2:1
DP/DI2:1
CP/DP2:1
CP/DI2:1
Water only Incubator conditions 41ºC dark
DP/DI2:1
CP/DP2:1
Microcosm Experiment 2
Water and sediment Algae room conditions 19ºC with
alternating light and dark cycles
CP
DPCP
CP
Microcosm Experiment 4
CPCP
Water and sediment Incubator conditions 41ºC dark
Water and sediment Algae room conditions 19ºC with
alternating light and dark cycles
CPCP
Microcosm Experiment 3
Water only Algae room conditions 19ºC with
alternating light and dark cycles
249250251252253254255
257258259260261262263264265266267268269270271272273274275276277278279
Supplementary Figure S2. Microcosm experiment 1: Water from all sites, chlorophyll and enterococci measured every 12 hours.
0 12 24 360
5
10
15
20
25
30
35
0
0.5
1
1.5
2
2.5
3
3.5
4
Hours
Chlo
roph
yll µ
g/L
Ente
roco
cci (
norm
alize
d)
Unfiltered FilteredSymbol Water Symbol Water
Chlorophyll a Lake Okeechobee Lake Okeechobee
Roosevelt Bridge Roosevelt BridgeDubois Park Dubois Park
Enterococci Lake Okeechobee Lake OkeechobeeRoosevelt Bridge Roosevelt Bridge
Dubois Park Dubois Park
280281282283284285
287288289
290291292293294295
0 12 24 360
2
4
6
8
10
12
14
0
0.2
0.4
0.6
0.8
1
1.2
Hours
Chlo
roph
yll a
µg/
L
Ente
roco
cci (
norm
alize
d)
Incubator Algae Culture RoomSymbol Dilution 1 Dilution 2 Symbol Dilution 1 Dilution 2
Chlorophyll a Lake Okeechobee
Deionized Water (DI)
Lake Okeechobee
DI
Dubois Park DI Dubois Park DI
Lake Okeechobee
Dubois Park Lake Okeechobee
Dubois Park
Enterococci Lake Okeechobee
DI Lake Okeechobee
DI
Not included in analysis
Dubois Park DI Dubois Park DI
Lake Okeechobee
Dubois Park Lake Okeechobee
Dubois Park
Supplementary Figure S3. Microcosm experiment 2: Water from Dubois Park and Lake Okeechobee, chlorophyll a and enterococci measured every 12 hours. Microcosms are 50:50 dilutions of Dilution 1 and Dilution 2 per table legend. Sample size n=1 per treatment.
296
297298299300301
References
American Public Health Association (APHA), American Water Works Association, Water Environment Federation. 2012. Standard Methods for the Examination of Water and Wastewater, 22nd Edition. E.W. Rice, R.B. Baird, A.D. Eaton, L.S. Clesceri, editors. da.
EPA Method 1600: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus Indoxyl-β-D-Glucoside Agar (mEI) December 2009 https://nepis.epa.gov Viewed 1/22/18
EPA Methods for the Determination of Inorganic Substances in Environmental Samples, EPA/600/R-09/100, August 1993 https://nepis.epa.gov Viewed 1/22/18
EPA Methods for the Determination of Inorganic Substances in Environmental Samples, EPA/600/R-09/100, Method 353.2: “Nitrogen, Nitrate-Nitrite” 1993 https://nepis.epa.gov Viewed 1/22/18
Florida Department of Environmental Protection Algal Bloom Sampling Results 2018 https://floridadep.gov/dear/algal-bloom/content/algal-bloom-sampling-results Viewed 3/16/19
IDEXX Laboratories, Inc., 2015, Enterolert and Quanti-Tray Technical Documents https://www.idexx.com/en/water/water-products-services/enterolert/ Viewed 11/13/18
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