Mazzei, Viviana Sullivan, Kristy 20220902 spreadsheet Lake Okeechobee HABs Project Nitrate, Ammonium, Phosphate Mesocosms Orlando, FL U.S. Geological Survey https://doi.org/10.5066/P99ELCEC An interdisciplinary and multiagency study was undertaken to study harmful algal bloom (HAB) dynamics on the Caloosahatchee River, which drains to the west from Lake Okeechobee into the Gulf of Mexico. Algae and cyanobacteria play crucial roles in aquatic ecosystems, but under favorable environmental conditions certain taxa may experience population growth booms resulting in HABs. Four independent mesocosm experiments were conducted in September 2019, June 2020, September 2020, and February 2021 to capture natural temporal variability in water quality and phytoplankton assemblages. The experimental installation consisted of three floating metal cradles each containing four fiberglass chambers filled with native river water. The twelve chambers were assigned to one of four treatments: phosphate (P), nitrate (N), ammonium (A), or control (C) – each with 3 replicates. The P, N, and A treatment chambers were enriched by applying a concentrated dosing solution of either dibasic dodecahydrate sodium phosphate, anhydrous sodium nitrate, or liquid ammonium hydroxide respectively to elevate concentrations above ambient levels. The mesocosms were treated and sampled over a 72-hour period (Time 0, Time 24, Time 48, Time 72). This dataset contains phytoplankton absolute and relative abundance measures at the genus level as natural units (NU/mL), cell densities (cells/mL), and biovolumes (um3/mL). Data were collected to understand the short-term effects of ammonium, nitrate, and phosphate enrichment independently on the phytoplankton assemblage of the Caloosahatchee River. 20190916 20190917 20190918 20190919 20200608 20200609 20200610 20200611 20200914 20200915 20200916 20200917 20210222 20210223 20210224 20210225 ground condition In work Irregular W.P. Franklin Lock and Dam -81.69897 -81.68738 26.72733 26.71732 USGS Thesaurus Algal blooms Eutrophication Field Experiments Phytoplankton USGS Metadata Identifier USGS:623de97ed34e915b67d6ae4b Caloosahatchee River Fort Myers, Florida W.P. Franklin Lock and Dam None Chlorophyta Actinastrum Coelastrum Crucigenia Scenedesmus Ankistrodesmus Monoraphidium Quadrigula Pediastrum Eudorina Pandorina Micractinium Schroederia Tetraedron Sphaerocystis Pyramichlamys Botryococcus Oocystis Dactylococcopsis Tetraselmis Arthrodesmus Cosmarium Staurastrum Closterium Elakatothrix Aphanizomenon Pseudanabaena Phormidium Oscillatoria Planktothrix Romeria Raphidiopsis Aphanocapsa Aphanothece Chroococcus Gomphosphaeria Merismopedia Microcystis Aulacoseira Skeletonema Cocconeis Cymbella Encyonema Gomphonema Navicula Nitzschia Fragilaria Ulnaria Chrysophyta Chrysosphaerella Mallomonas Synura Dinobryon Cryptomonas Rhodomonas Gymnodiniales Peridiniales Haptophyta Euglena Lepocinclis Phacus Strombomonas Trachelomonas Anabaenopsis Ceratium Life Life Kingdom Plantae Subkingdom Viridiplantae Infrakingdom Chlorophyta Division Chlorophyta TSN: 5414 Subdivision Chlorophytina Class Chlorophyceae Order Sphaeropleales Family Scenedesmaceae Genus Actinastrum TSN: 6098 Genus Coelastrum TSN: 6273 Genus Crucigenia TSN: 6225 Genus Scenedesmus TSN: 5414 Family Selenastraceae Genus Ankistrodesmus TSN: 5877 Genus Monoraphidium TSN: 5990 Genus Quadrigula TSN: 5938 Family Hydrodictyaceae Genus Pediastrum TSN: 6031 Order Chlamydomonadales Family Volvocaceae Genus Eudorina TSN: 5597 Genus Pandorina TSN: 5578 Family Micractiniaceae Genus Micractinium TSN: 6321 Family Chlorococcaceae Genus Schroederia TSN: 5656 Genus Tetraedron TSN: 5661 Family Sphaerocystidaceae Genus Sphaerocystis TSN: 9169 Family Chlamydomonadaceae Genus Pyramichlamys TSN: 5520 Class Trebouxiophyceae Order Trebouxiales Family Botryococcaceae Genus Botryococcus TSN: 6306 Order Chlorellales Family Oocystaceae Genus Oocystis TSN: 5827 Class Ulvophyceae Order Ulotrichales Family Ulotrichaceae Genus Dactylococcopsis TSN: 6446 Class Chlorodendrophyceae Order Chlorodendrales Family Chlorodendraceae Genus Tetraselmis TSN: 9560 Infrakingdom Streptophyta Superdivision Charophyta Division Charophyta Class Conjugatophyceae Order Desmidiales Family Desmidiaceae Genus Arthrodesmus TSN: 8276 Genus Cosmarium TSN: 7848 Genus Staurastrum TSN: 7440 Family Closteriaceae Genus Closterium TSN: 7257 Class Klebsormidiophyceae Order Klebsormidiales Family Elakatotrichaceae Genus Elakatothrix TSN: 9412 Kingdom Bacteria Subkingdom Negibacteria Phylum Cyanobacteria Class Cyanophyceae Order Nostocales Family Nostocaceae Genus Aphanizomenon TSN: 1191 Genus Pseudanabaena TSN: 1175 Genus Anabaenopsis TSN: 1182 Family Oscillatoriaceae Genus Phormidium TSN: 992 Genus Oscillatoria TSN: 917 Genus Planktothrix TSN: 189420 Genus Romeria TSN: 1078 Family Rivulariaceae Genus Raphidiopsis TSN: 1290 Order Chroococcales Family Chroococcaceae Genus Aphanocapsa TSN: 625 Genus Aphanothece TSN: 636 Genus Chroococcus TSN: 654 Genus Gomphosphaeria TSN: 714 Genus Merismopedia TSN: 727 Genus Microcystis TSN: 747 Kingdom Chromista Subkingdom Harosa Infrakingdom Halvaria Superdivision Heterokonta Division Ochrophyta Class Bacillariophyceae Subclass Coscinodiscophycidae Order Aulacoseirales Family Aulacoseiraceae Genus Aulacoseira TSN: 590863 Order Thalassiosirales Family Skeletonemataceae Genus Skeletonema TSN: 2401 Subclass Bacillariophycidae Order Achnanthales Family Cocconeidaceae Genus Cocconeis TSN: 3577 Order Cymbellales Family Cymbellaceae Genus Cymbella TSN: 4795 Genus Encyonema TSN: 590838 Family Gomphonemataceae Genus Gomphonema TSN: 4911 Order Naviculales Family Naviculaceae Genus Navicula TSN: 3649 Order Bacillariales Family Bacillariaceae Genus Nitzschia TSN: 5070 Subclass Fragilariophycidae Order Fragilariales Family Fragilariaceae Genus Fragilaria TSN: 2932 Genus Ulnaria TSN: 970000 Subkingdom Chromista Division Chrysophyta TSN: 1447 Class Chrysophyceae Order Ochromonadales Family Synuraceae Genus Chrysosphaerella TSN: 1669 Genus Mallomonas TSN: 1598 Genus Synura TSN: 1655 Family Dinobryaceae Genus Dinobryon TSN: 1515 Division Cryptophycophyta Class Cryptophyceae Order Cryptomonadales Family Cryptomonadaceae Genus Cryptomonas TSN: 10635 Genus Rhodomonas TSN: 10663 Division Pyrrophycophyta Class Dinophyceae Order Gymnodiniales TSN: 9994 Order Peridiniales TSN: 9995 Order Gonyaulacales Family Ceratiaceae Genus Ceratium TSN: 10397 Division Haptophyta TSN: 2134 Kingdom Protozoa Phylum Euglenophycota Class Euglenophyceae Order Euglenales Family Euglenaceae Genus Euglena TSN: 9620 Genus Lepocinclis TSN: 9758 Genus Phacus TSN: 9766 Genus Strombomonas TSN: 9740 Genus Trachelomonas TSN: 9690 none none Viviana Mazzei SOUTHEAST REGION: CARIBBEAN-FLORIDA WSC STUDIES SEC Research Biologist mailing and physical

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Orlando FL 32826 US 407-803-5508 vmazzei@usgs.gov U.S. Army Corps of Engineers, Nova Southeastern University, Florida Gulf Coast University Data were analyzed by a certified aquatic laboratory (PhycoTech, Inc.) that completes it's own quality assurance and quality control procedures. All data were broadly checked for accuracy with no obvious outliers present in the data. No formal logical accuracy tests were conducted Data set is considered complete for the information presented, as described in the abstract. Users are advised to read the rest of the metadata record carefully for additional details. No formal horizontal accuracy tests were conducted No formal vertical accuracy tests were conducted The experimental installation consisted of three floating metal cradles each containing four 556-liter fiberglass chambers filled with native river water using a gas-powered pump. The twelve chambers were assigned to one of four treatments, phosphate (P), nitrate (N), ammonium (A), or control (C) – each with 3 replicates. The P, N, and A treatment chambers were enriched by applying a concentrated dosing solution of either dibasic dodecahydrate sodium phosphate (Na2HPO4·12H2O), anhydrous sodium nitrate (NaNO3), or liquid ammonium hydroxide (NH4OH) respectively to elevate concentrations above ambient levels. Nutrient dosing solutions were prepared as follows: for the A treatment, liquid ammonium hydroxide (NH4OH) was combined with 100 mL of deionized (DI) water to the desired concentration; for the N treatment, anhydrous sodium nitrate (NaNO3) was dissolved in 100 mL of DI water to the desired concentration; and for the P treatment, 98.5% dibasic dodecahydrate sodium phosphate (Na2HPO4·12H2O was dissolved in 100 mL of DI water to the desired concentration. The mesocosms were treated and sampled using a surface water grab at approximately 24-hour intervals over a 72-hour period (Time 0, Time 24, Time 48, Time 72). Actual time of collection was not recorded, but samples were generally collected between 7 am to 4 pm, local time. Upon collection, live and 1 percent preserved glutaraldehyde samples were shipped on ice to Phycotech Inc. for analysis. 2021 The phytoplankton community structure for each sample was characterized using an Imaging FlowCytobot (IFCB, McLane Research Labs) for taxa classification and enumeration. Live samples were shipped overnight and run through the IFCB which captures images of particulates between 2-250 µm as they pass through a flow cell. The particulates were identified by an automated classifier which identifies algal units above 10 µm. The automated algal classifications made by the IFCB were validated by a phycologist (Ann St. Amand, PhD). Results from the IFCB are semi-quantitative and are most informative when describing dominant taxa and functional groups. The images and image counts were used to provide estimates of relative and absolute algal concentration and biovolume for the recorded sample volume that was analyzed. Estimates of algal concentration and biovolumes were procured from the IFCB images and data were delivered to USGS in an excel file/CSV format. This dataset contains phytoplankton absolute and relative abundance measures at the genus level as natural units per milliliter (NU/mL), cell densities per milliliter (cells/mL), and biovolumes in cubic microns per milliliter (um3/mL) with additional functional group classification. 2021 Phycotech Inc. Ann St. Amand President/Owner mailing and physical

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St. Joseph MI 49085 USA 2699833654 2699833653 astamand@phycotech.com 0.00001 0.00001 Decimal degrees Phytoplankton Counts Identification and enumeration of phytoplankton taxa at the genus level Producer-defined SampleUniqueID A unique alphanumeric identifier for each sample Producer-defined An automatically generated internal sample identifier consisting of 16 characters starting with the letter D followed by the 8-digit date (YYYYMMDD) the sample was analyzed and the letter T followed by the 6-digit time (Greenwich Mean Time; 24 hour time) the sample file was generated Treatment The experimental treatment chamber the sample was collected from User-defined C-1 Control treatment, replicate 1 of 3 Producer-defined C-2 Control treatment, replicate 2 of 3 Producer-defined C-3 Control treatment, replicate 3 of 3 Producer-defined P-1 Phosphate treatment, replicate 1 of 3 Producer-defined P-2 Phosphate treatment, replicate 2 of 3 Producer-defined P-3 Phosphate treatment, replicate 3 of 3 Producer-defined N-1 Nitrate treatment, replicate 1 of 3 Producer-defined N-2 Nitrate treatment, replicate 2 of 3 Producer-defined N-3 Nitrate treatment, replicate 3 of 3 Producer-defined A-1 Ammonium treatment, replicate 1 of 3 Producer-defined A-2 Ammonium treatment, replicate 2 of 3 Producer-defined A-3 Ammonium treatment, replicate 3 of 3 Producer-defined Time The collection time relative to approximate hours from the start of each mesocosm experiment, starting at time 0 hours and ending at time 72 hours. Producer-defined T-0 Time 0 - initial sample collection before nutrient dosing Producer-defined T-24 Time 24 - sample collection at 24 hours Producer-defined T-48 Time 48 - sample collection at 48 hours Producer-defined T-72 Time 72 - sample collection at 72 hours Producer-defined DateSampled The date the sample was collected in month/day/year format Producer-defined 9/16/2019 2/25/2021 MM/DD/YY VolumeAnalyzed_mL Volume of sample in milliliters run through the Imaging FlowCytobot for analysis. Producer Defined 0.0681 3.5938 milliliter 0.0001 Preservative Notes whether sample was counted live or preserved with 1 percent glutaraldehyde before being run through the Imaging FlowCytobot. Producer Defined Live Live sample analyzed Producer defined Glutaraldehyde 1% glutaraldehyde sample analyzed Producer defined Taxon Lowest taxonomic group of an identified algae Producer Defined One of the identified algae taxa or groups found in the mesocosm sample. FunctionalGroup The ecological functional group of the taxa Producer-defined BG Cyanobacteria that do not produce toxins or taste and odor compounds Producer-defined TO Taste and Odor producing cyanobacteria Producer-defined CP Cryptophytes and Dinoflagellates Producer-defined DY Chrysophytes, Haptophytes, and Diatoms Producer-defined E Euglenophytes Producer-defined G Chlorophytes Producer-defined HAB Toxin-producing cyanobacteria Producer-defined U Unclassified - Images that the classifier cannot confidently identify. Includes small flagellates entrained in detritus, taxa not yet included in the classifier, partial images and images with multiple taxa. Producer-defined M Miscellaneous - All other groups including small chlorophytes or cyanobacteria less than 9um in diameter Producer-defined CER Dinoflagellate of the genus Ceratium Producer defined NUcount The number of natural units (colonies or filaments) counted in a sample. Data are not rounded to USGS significant figures. Producer-defined 1 1068 Natural units CellCount The number of cells counted in a sample. Data are not rounded to USGS significant figures. Producer-defined 1.0 9760.0 Cells Mean_Cells_Per_NU The mean number of cells per natural unit. Data are not rounded to USGS significant figures. Producer-defined 1.0 253.7 cells per natural unit NU_Density_mL The total number of natural units in 1 milliliter of sample. Data are not rounded to USGS significant figures. Producer-defined 0.3 9632.4 Natural units per milliliter Relative_NU Relative natural units per milliliter of sample. The proportion of natural units of a taxon relative to the total number of natural units of all taxa in 1 milliliter of sample. Data are not rounded to USGS significant figures. Producer-defined 0.0004 0.5494 Cell_Density_mL Absolute cell abundance. The total number of cells in 1 milliliter of sample. Data are not rounded to USGS significant figures. Producer-defined 0.3 49572.9 Cells per milliliter Relative_Cells Relative cell abundance. The proportion of cells of a taxon relative to total number of cells of all taxa in 1 milliliter of sample. Data are not rounded to USGS significant figures. Producer-defined 0.0001 0.7737 IFCB_Biovolume_um3_per_mL Cell volume per milliliter of sample. The volume of cells in cubic microns per 1 milliliter of sample calculated by an iterative algorithm based on over a hundred dimensions for each image taken by the Imaging FLowCytobot (IFCB). Data are not rounded to USGS significant figures. Producer-defined 25 6898212 Cubic micrometer per milliliter Relative_Biovolume Relative Cell Volume (unitless). The ratio of volume occupied by a taxon relative to the volume occupied by all taxa in 1 milliliter of sample. Data are not rounded to USGS significant figures. Producer-defined 0.00002 0.66231 Mean_IFCB_GALD_um Greatest axial linear dimension in micrometers. The longest way you can measure the natural unit. Data are not rounded to USGS significant figures. Producer-defined 7.8 389.0 Micrometer U.S. Geological Survey - ScienceBase mailing and physical

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Denver CO 80225 USA 1-888-275-8747 sciencebase@usgs.gov Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the U.S. Government. Although this information product, for the most part, is in the public domain, it also contains copyrighted materials as noted in the text. Permission to reproduce copyrighted items must be secured from the copyright owner whenever applicable. The data have been approved for release and publication by the USGS. Although the data have been subjected to rigorous review and are substantially complete, the USGS reserves the right to revise the data pursuant to further analysis and review. Furthermore, the data are released on the condition that neither the USGS nor the U.S. Government may be held liable for any damages resulting from authorized or unauthorized use. Although the data have been processed successfully on a computer system at the USGS, no warranty expressed or implied is made regarding the display or utility of the data on any other system, or for general or scientific purposes, nor shall the act of distribution constitute any such warranty. The USGS shall not be held liable for improper or incorrect use of the data described and/or contained herein. Users of the data are advised to read all metadata and associated documentation thoroughly to understand appropriate use and data limitations. Digital Data https://doi.org/10.5066/P99ELCEC None 20220902 Viviana Mazzei USGS Research Biologist mailing and physical

12703 Research Parkway

Orlando Florida 32826 USA 407-803-5508 vmazzei@gmail.com FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata FGDC-STD-001.1-1999