Initial estuarine response to inorganic nutrient inputs from a legacy mining facility adjacent to Tampa Bay, Florida.

Legacy mining facilities pose significant risks to aquatic resources. From March 30th to April 9th, 2021, 814 million liters of phosphate mining wastewater and marine dredge water from the Piney Point facility were released into lower Tampa Bay (Florida, USA). This resulted in an estimated addition of 186 metric tons of total nitrogen, exceeding typical annual external nitrogen load estimates to lower Tampa Bay in a matter of days. An initial phytoplankton bloom (non-harmful diatoms) was first observed in April. Filamentous cyanobacteria blooms (Dapis spp.) peaked in June, followed by a bloom of the red tide organism Karenia brevis. Reported fish kills tracked K. brevis concentrations, prompting cleanup of over 1600 metric tons of dead fish. Seagrasses had minimal changes over the study period. By comparing these results to baseline environmental monitoring data, we demonstrate adverse water quality changes in response to abnormally high and rapidly delivered nitrogen loads.

Effects of Zinc in an Outdoor Freshwater Microcosm System.

A long-term exposure outdoor microcosm study was conducted to evaluate the effects of zinc (Zn) on zooplankton, phytoplankton, and periphyton in a freshwater system. Five Zn treatment concentrations (nominal: 8, 20, 40, 80, and 160 µg/L Zn) and an untreated control with 3 replicates each were used. Various physical and chemical characteristics of the microcosms and biological assessment endpoints (e.g., total abundance, group abundance, species richness, chlorophyll a, etc.) were measured to determine the effects of Zn over time. In general, physical and chemical characteristics (e.g., total dissolved solids, total suspended solids, dissolved oxygen, pH, dissolved organic carbon) of water fluctuated over time, but they were not significantly different within treatments and controls during the study. Zinc significantly affected the population dynamics and community structure of plankton. The effects occurred 7 d after initial treatment exposures began and continued to the end of the treatment phase, especially at the high treatment concentrations. Total and group abundance, species richness, the Shannon index, and chlorophyll a concentrations for high Zn treatment concentrations were significantly lower than the controls during the treatment phase. The no-observed-effect, lowest-observed-effect, and median effect concentrations were generally lower than the literature-reported results from single-species toxicity tests for fish and invertebrates, suggesting that plankton are more sensitive to Zn than planktivores. Although primary producers play an important role in the ecosystem, they have not been consistently incorporated into numerical environmental quality criteria for freshwater organisms, at least in the United States. The results of the present study are useful for development of environmental quality guidelines for freshwater ecosystems and ecological risk assessment. Environ Toxicol Chem 2021;40:2053-2072. © 2021 SETAC.

High-resolution Spatiotemporal Dynamics of Harmful Algae in the Indian River Lagoon (Florida)-A Case Study of Aureoumbra lagunensis, Pyrodinium bahamense, and Pseudo-nitzschia.

The Indian River Lagoon (IRL), located on the east coast of Florida, is a complex estuarine ecosystem that is negatively affected by recurring harmful algal blooms (HABs) from distinct taxonomic/functional groups. Enhanced monitoring was established to facilitate rapid quantification of three recurrent bloom taxa, Aureoumbra lagunensis, Pyrodinium bahamense, and Pseudo-nitzschia spp., and included corroborating techniques to improve the identification of small-celled nanoplankton (<10 µm in diameter). Identification and enumeration of these target taxa were conducted during 2015-2020 using a combination of light microscopy and species-specific approaches, specifically immunofluorescence flow cytometry as well as a newly developed qPCR assay for A. lagunensis presented here for the first time. An annual bloom index (ABI) was established for each taxon based on occurrence and abundance data. Blooms of A. lagunensis (>2×108 cells L-1) were observed in all six years sampled and across multiple seasons. In contrast, abundance of P. bahamense, largely driven by the annual temperature cycle that moderates life cycle transitions and growth, displayed a strong seasonal pattern with blooms (105-107 cells L-1) generally developing in early summer and subsiding in autumn. However, P. bahamense bloom development was delayed and abundance was significantly lower in years and locations with sustained A. lagunensis blooms. Pseudo-nitzschia spp. were broadly distributed with sporadic bloom concentrations (reaching 107 cells L-1), but with minimal concentrations of the toxin domoic acid detected (<0.02 µg L-1). In summer 2020, multiple monitoring tools characterized a novel nano-cyanobacterium bloom (reaching 109 cells L-1) that coincided with a decline in A. lagunensis and persisted into autumn. Statistical and time-series analyses of this spatiotemporally intensive dataset highlight prominent patterns in variability for some taxa, but also identifies challenges of characterizing mechanisms underlying more episodic yet persistent events. Nevertheless, the intersect of temperature and salinity as environmental proxies proved to be informative in delineating niche partitioning, not only in the case of taxa with long-standing data sets but also for seemingly unprecedented blooms of novel nanoplanktonic taxa.

Hurricanes, El Niño and harmful algal blooms in two sub-tropical Florida estuaries: Direct and indirect impacts.

Future increases in the intensity of hurricanes and El Niño periods predicted by climate change models have focused attention on their role in stimulating harmful algal blooms (HABs). A series of hurricanes that recently impacted Florida (USA) provided a unique opportunity to explore the relationships between hurricanes, El Niño and HABs in two Florida estuaries subject to repeated intense ecosystem disruptive HABs, the Indian River Lagoon and the St. Lucie Estuary. The roles that hurricanes and El Niño play in contributing to HAB events are examined in the context of key structural and functional features of each estuary and their watersheds, including morphology, water residence time and hydrology, such as the influence of Lake Okeechobee discharges into the St. Lucie Estuary. The most direct impact was the increase in rainfall associated with hurricanes and El Niño, resulting in enhanced nutrient loads which drive HABs in the Indian River Lagoon and Lake Okeechobee. Major HABs in Lake Okeechobee also present an indirect threat of freshwater HAB blooms in the St. Lucie Estuary via mandated discharges from the lake into the estuary during high rainfall periods. Conversely, during the absence of HABs in Lake Okeechobee, short water residence times produced by discharges into the St. Lucie Estuary can result in lower bloom intensities.

Revealing Biotic and Abiotic Controls of Harmful Algal Blooms in a Shallow Subtropical Lake through Statistical Machine Learning.

Harmful algal blooms are a growing human and environmental health hazard globally. Eco-physiological diversity of the cyanobacteria genera that make up these blooms creates challenges for water managers tasked with controlling the intensity and frequency of blooms, particularly of harmful taxa (e.g., toxin producers, N2 fixers). Compounding these challenges is the ongoing debate over the efficacy of nutrient management strategies (phosphorus-only versus nitrogen and phosphorus), which increases decision-making uncertainty. To improve our understanding of how different cyanobacteria respond to nutrient levels and other biophysical factors, we analyzed a unique 17 year data set comprising monthly observations of cyanobacteria genera and zooplankton abundances, water quality, and flow in a bloom-impacted, subtropical, flow-through lake in Florida (United States). Using the Random Forests machine learning algorithm, an ensemble modeling approach, we characterized and quantified relationships among environmental conditions and five dominant cyanobacteria genera. Results highlighted nonlinear relationships and critical thresholds between cyanobacteria genera and environmental covariates, the potential for hydrology and temperature to limit the efficacy of cyanobacteria bloom management actions, and the importance of a dual nutrient management strategy for reducing bloom risk in the long term.

Investigation of extraction and analysis techniques for Lyngbya wollei derived Paralytic Shellfish Toxins.

Paralytic Shellfish Toxins (PSTs) are highly toxic metabolic by-products of cyanobacteria and dinoflagellates. The filamentous cyanobacterium Lyngbya wollei produces a unique set of PSTs, including L. wollei toxins (LWT) 1-6. The accurate identification and quantification of PSTs from Lyngbya filaments is challenging, but critical for understanding toxin production and associated risk, as well as for providing baseline information regarding the potential for trophic transfer. This study evaluated several approaches for the extraction and analysis of PSTs from field-collected L. wollei dominated algal mats. Extraction of PSTs from lyophilized Lyngbya biomass was assessed utilizing hydrochloric acid and acetic acid at concentrations of 0.001-0.1 M. Toxin profiles were then compared utilizing two analysis techniques: pre-column oxidation (peroxide and periodate) High Performance Liquid Chromatography (HPLC) with Fluorescence (FL) detection and LC coupled with Mass Spectrometry (MS). While both acid approaches efficiently extracted PSTs, hydrochloric acid was found to convert the less toxic LWT into the more toxic decarbamoylgonyautoxins 2&3 (dcGTX2&3) and decarbamoylsaxitoxin (dcSTX). In comparison, extraction with 0.1 M acetic acid preserved the original toxin profile and limited the presence of interfering co-extractants. Although pre-chromatographic oxidation with HPLC/FL was relatively easy to setup and utilize, the method did not resolve the individual constituents of the L. wollei derived PST profile. The LC/MS method allowed characterization of the PSTs derived from L. wollei, but without commercially available LWT 1-6 standards, quantitation was not possible for the LWT. In future work, evaluation of the risk associated with L. wollei derived PSTs will require commercially available standards of LWT 1-6 for accurate determinations of total PST content and potency.

Aquatic biosystems: reactions and actions.

Aquatic biological systems are a critical part of the structure and function of earth's biosphere. While attention of the scientific community is often focused on the reaction of biological systems to changes in the environment, these systems also have profound effects, or actions, on the environment. Throughout the evolutionary history of earth, the rise and/or fall of different aquatic biosystems has impacted the character of the biosphere. At no time have environmental changes been more important to all life on earth than in the modern era, which underscores the need for the new journal, Aquatic Biosystems. We welcome submission of original research manuscripts, reviews, and commentaries to the journal.

Diversity of and selection acting on cylindrospermopsin cyrB gene adenylation domain sequences in Florida.

Aphanizomenon ovalisporum is the only confirmed cylindrospermopsin producer identified in the United States to date. On the other hand, Cylindrospermopsis raciborskii is a prominent feature of many lakes in Florida and other regions of the United States. To see the variation in cylindrospermopsin cyrB gene adenylation domain sequences and possibly discover new cylindrospermopsin producers, we collected water samples for a 3-year period from 17 different systems in Florida. Positive amplicons were cloned and sequenced, revealing that approximately 92% of sequences were A. ovalisporum-like (>99% identity). Interestingly, 6% of sequences were very similar (>99% identity) to cyrB sequences of C. raciborskii from Australia and of Aphanizomenon sp. from Germany. Neutrality tests suggest that A. ovalisporum-like cyrB adenylation domain sequences are under purifying selection, with abundant low-frequency polymorphisms within the population. On the other hand, when compared between species by codon-based methods, amino acids of CyrB also seem to be under purifying selection, in accordance with the one proposed amino acid thought to be activated by the CyrB adenylation domain.

A new attachment device for deployment of monitoring equipment in estuaries and other high-energy environments.

Biomonitoring is an important component of estuarine research and monitoring programs because living organisms integrate biological, chemical, and physical conditions over time. The deployment of biomonitoring devices in ecosystems that are subject to changes in water level and flow can be very challenging. This paper describes a new device, which facilitates such applications such as the deployment of periphytometers. The device is designed to encircle posts, poles, or pilings, such as channel markers common in many waterways. This device has been evaluated and approved for use by the US Coast Guard, needed for attachment to navigational aids. It allows attachment of monitoring devices requiring in situ deployment at fixed water depths in systems with dynamic water levels or velocities while minimizing the potential for shading, damage, theft, or poor long-term performance.

Temporal and spatial variations of copper, cadmium, lead, and zinc in Ten Mile Creek in South Florida, USA.

Lead (Pb), zinc (Zn), copper (Cu), and cadmium (Cd) often seriously deteriorate water quality. Spatial and temporal fluctuations of the metal concentrations in the Ten Mile Creek (Florida) (TMC) were monitored on a weekly basis at 7 sampling sites, from June 2005 to September 2007. River sediment samples were also collected from these sites in April, June, and October 2006 and January 2007, and analyzed for water, Mehlich 1 (M1), and Mehlich 3 (M3)-extractable metals (Mehlich, 1953, 1984), to examine the role of sediments as sources or sinks of the metals. The concentrations of lead, zinc, copper, and cadmium in the water samples were

IMPROVED METHODS FOR THE ISOLATION OF CYANOBACTERIAL DNA FROM ENVIRONMENTAL SAMPLES(1).

DNA isolated from environmental samples often contains enzyme inhibitors disruptive to downstream molecular applications. Most of the existing methods of cyanobacterial DNA isolation do not effectively eliminate these inhibitors from sediment samples or cells collected from freshwater ecosystems. We describe improved methods based on the xanthogenate-SDS nucleic acid isolation (XS) method of Tillett and Neilan (2000). Our improved methods provided high-quality cyanobacterial DNA that could be amplified in PCR and digested with a restriction enzyme. Results were superior to several commercial kits. The DNA yield was also similar to that obtained via the standard XS method. These methods should provide valuable new tools for the expanded application of molecular genetics to limnological and oceanographic research.

Debromoaplysiatoxin in Lyngbya-dominated mats on manatees (Trichechus manatus latirostris) in the Florida King's Bay ecosystem.

Proliferation of the potentially toxic cyanobacterium, Lyngbya, in Florida lakes and rivers has raised concerns about ecosystem and human health. Debromoaplysiatoxin (DAT) was measured in concentrations up to 6.31 microg/g wet weight lyngbyatoxin A equivalents (WWLAE) in Lyngbya-dominated mats collected from natural substrates. DAT was also detected (up to 1.19 microg/g WWLAE) in Lyngbya-dominated mats collected from manatee dorsa. Ulcerative dermatitis found on manatees is associated with, but has not been proven to be caused by DAT.

Temporal and spatial variations of nutrients in the Ten Mile Creek of South Florida, USA and effects on phytoplankton biomass.

Water quality throughout south Florida has been a major concern for many years. Nutrient enrichment in the Indian River Lagoon (IRL) is a major surface water issue and is suggested as a possible cause of symptoms of ecological degradation. In 2005-06, water samples were collected weekly from seven sites along Ten Mile Creek (TMC), which drains into the Indian River Lagoon, to investigate and analyze spatial and temporal fluctuations of nutrients nitrogen (N) and phosphorus (P). The objective of this study was to understand the relationships among chlorophyll a concentration, nutrient enrichment and hydrological parameters in the surface water body. High median concentrations of total P (TP, 0.272 mg L(-1)), PO4-P (0.122 mg L(-1)), and dissolved total P (DTP, 0.179 mg L(-1)); and total N (TN, 0.988 mg L(-1)), NO3(-)-N (0.104 mg L(-1)), NH4+-N (0.103 mg L(-1)), and total Kjeldahl N (TKN, 0.829 mg L(-1)), were measured in TMC. The concentrations of TP, PO4-P, DTP, TN, NO3(-)-N, NH4+-N, and TKN were higher in summer and fall than in winter and spring. However, chlorophyll a and pheophytin concentrations during this period in TMC varied in the range of 0.000-60.7 and 0.000-17.4 microg L(-1), with their median values of 3.54 and 3.02 microg L(-1), respectively. The greatest mean chlorophyll a (10.3 microg L(-1)) and pheophytin (5.71 microg L(-1)) concentrations occurred in spring, while the lowest chlorophyll a (1.49 microg L(-1)) and pheophytin (1.97 mug L(-1)) in fall. High concentrations of PO4-P (>0.16 mg L(-1)), DTP (>0.24 mg L(-1)), NO3(-)-N (>0.15 mg L(-1)), NH4+-N (>0.12 mg L(-1)), and TKN (>0.96 mg L(-1)), occurred in the upstream of TMC, while high concentrations of chlorophyll a (>6.8 mug L(-l)) and pheophytin (>3.9 microg L(-l)) were detected in the downstream of TMC. The highest chlorophyll a (11.8 mug L(-l)) and pheophytin (6.06 microg L(-l)) concentrations, however, were associated with static and open water conditions. Hydrological parameters (total dissolved solid, electrical conductivity, salinity, pH, and water temperature) were positively correlated with chlorophyll a and pheophytin concentrations (P < 0.01) and these factors overshadowed the relationships between N and P concentrations and chlorophyll a under field conditions. Principal component analysis and the ratios of DIN/DP and TN/TP in the water suggest that N is the limiting nutrient factor for phytoplankton growth in the TMC and elevated N relative to P is beneficial to the growth of phytoplankton, which is supported by laboratory culture experiments under controlled conditions.

A comparative study of Florida strains of Cylindrospermopsis and Aphanizomenon for cylindrospermopsin production.

The toxin cylindrospermopsin (CYN) is produced by a variety of cyanobacterial genera. One of these, Cylindrospermopsis raciborskii, is generally assumed to be the source of CYN in lakes and rivers in Florida, USA. However, in this study, none of the eight Florida isolates of this species tested contained the genetic determinants involved in toxin production nor did they produce CYN. We show for the first time that Aphanizomenon ovalisporum isolated from a pond in this state has the genes putatively associated with CYN production. Analysis by liquid chromatography with mass spectrometric detection (LC/MS) revealed that it produced CYN in the range of 7.39-9.33 microg mg(-1) freeze-dried cells. 16S rDNA sequences of this strain showed 99.6% and 99.9% identity to published A. ovalisporum and Anabaena bergii 16S sequences, respectively. These results help to explain the general lack of a defined relationship between the abundance of C. raciborskii in freshwater ecosystems of Florida and observed concentrations of CYN. The latter observation raises the potential that previous reports of CYN may be coincidental with unrecorded presence of another CYN-producing species.

Kinetics and uptake mechanisms for monomethylmercury between freshwater algae and water.

Uptake kinetics of monomethylmercury chloride (MeHgCl) were measured for two species of green algae (Selenastrum capricomutum and Cosmarium botrytis), one blue-green algae (Schizothrix calcicola), and one diatom (Thalassiosira weissflogii), algal species that are commonly found in natural surface waters. Species differences were found with the two green algae giving the highest uptake rates, and one of them (Cosmarium) showing differences between cultures having widely different cell age (exponential versus stationary), where increases in uptake rate for cells 30 days old were about 25 times greater than cells only 3 days old when weights of cells were considered. Both Schizothrix and Thalassiosira exhibited nearly the same lower uptake rates, approximately 20 times lower than the two green algal species. Experiments with photosystem inhibitors, uncouplers, gamma-radiation, light deprivation, and extended range uptake all point to an active transport mechanism for MeHgCl.