Efficacy of algaecides for the proactive treatment of overwintering cyanobacteria.

Once established within a water resource, harmful algal blooms (HABs) can occur seasonally with an intense and rapid onset, giving water resource managers limited time to respond to lessen risks. An attractive strategy to decrease human, ecological, and economic risks from HABs is to implement proactive algaecide treatments applied to overwintering cyanobacteria (i.e., akinetes and quiescent vegetative cells) in sediments prior to the formation of a HAB; however, this approach is novel and very limited efficacy data exist. Therefore, the specific objectives of this research were to 1) evaluate copper- and peroxide-based algaecides, applied as single and repeat treatments at the bench scale, to identify effective proactive treatments, and 2) compare correlations between cell density and other response measurements (i.e., in vivo chlorophyll a and phycocyanin concentrations and percent benthic coverage), to identify informative metrics to assess overwintering cyanobacteria responses. Twelve treatment scenarios using copper- and peroxide-based algaecides were applied to sediments containing overwintering cyanobacteria prior to a 14 d incubation under favorable growth conditions. Responses of cyanobacteria in the planktonic (i.e., cell density, in vivo chlorophyll a and phycocyanin concentrations) and benthic (percent coverage) phases after a 14 d incubation were evaluated in treatments and controls. The HAB-forming cyanobacteria present after a 14 d incubation were: Aphanizomenon, Dolichospermum, Microcystis, Nostoc, and Planktonthrix. Successive treatments of copper sulfate (CuSulfate) followed by sodium carbonate peroxyhydrate (PeroxiSolid) (second algaecide applied after 24 h) as well as repeat applications of a single algaecide, PeroxiSolid (second treatment applied after 24 h) resulted in statistically significant (p ≤ 0.05; α = 0.05) declines in cell density relative to untreated controls. Planktonic cyanobacteria responses measured in terms of phycocyanin concentrations were strongly correlated with cyanobacteria density measurements (Pearson's correlation coefficient (r) = 0.89). Chlorophyll a concentrations and percent benthic coverage did not correlate with planktonic cyanobacteria density measurements (r = 0.37 and -0.49, respectively) and therefore, were unreliable metrics for cyanobacterial responses in this study. These data provide initial evidence of the efficacy of algaecides for treating overwintering cells in sediments and contribute to our overarching hypothesis that proactive treatments may delay the onset and intensity of HABs in impacted waterbodies.

Laboratory-scale evaluation of algaecide effectiveness for control of microcystin-producing cyanobacteria from Lake Okeechobee, Florida (USA).

Growth of microcystin-producing cyanobacteria in Lake Okeechobee (Florida, USA) and surrounding waters has resulted in adverse health impacts for humans and endangered species, as well as significant economic losses. As these issues worsen, there is growing pressure for efficacious solutions to rapidly mitigate harmful algal blooms (HABs) and protect critical freshwater resources. Applications of USEPA-registered algaecides as management tactics meet many decision-making criteria often required by water resource managers (e.g., effective, scalable, selective), but have not yet been evaluated on a large scale within the Lake Okeechobee waterway. This study was conducted to bolster the peer-reviewed database for available management tactics against microcystin-producing cyanobacteria in waters of this region. Laboratory-scale experiments can be conducted first to minimize uncertainty at larger scales and improve confidence in decision-making. In this study, samples containing microcystin-producing cyanobacteria collected from Lake Okeechobee were exposed to several USEPA-registered algaecides in laboratory toxicity experiments. Responses of target cyanobacteria were measured 3 days after treatment (DAT) in terms of cell density, chlorophyll-a concentrations, and phycocyanin concentrations. Based on responses of the cyanobacteria, minimum effective exposure concentrations were identified for each algaecide. Microcystin release (i.e. proportion of total microcystins in the aqueous phase) was measured and compared 1 DAT among effective exposures. Total microcystin concentrations were measured in effective treatments at 1, 4, and 9 DAT to discern potential for microcystin persistence following exposures to the effective formulations and exposure concentrations. Overall, several formulations including GreenClean Liquid® 5.0, GreenClean Liquid® 5.0 combined with Hydrothol® 191, and the copper-based algaecides evaluated (Algimycin® PWF, Argos, Captain® XTR, Cutrine® Ultra, and SeClear®) achieved significant and similar effects on target cyanobacteria. The chelated copper-based formulations (Algimycin® PWF, Argos, Captain® XTR, and Cutrine® Ultra) resulted in relatively less microcystin release 1 DAT and lesser total microcystin concentrations 4 DAT. At 9 DAT, total microcystin concentrations were significantly lower than in untreated controls in all treatments evaluated. These results provide the necessary comparative performance data for preliminary decision-making and designing additional studies at larger scales. Importantly, the comparative toxicity data and approach provided in this study demonstrate the initial steps for development of site-specific management strategies for Lake Okeechobee and other areas impacted by harmful algal blooms with large spatial and temporal scales.

Relationship among aqueous copper half-lives and responses of Pimephales promelas to a series of copper sulfate pentahydrate concentrations.

Copper algaecide exposures in situ are often of shorter duration than exposures for static toxicity experiments because aqueous concentrations in situ dissipate as a function of site-specific fate processes. Consequently, responses of organisms to static copper exposures may overestimate effects following in situ exposures. To understand the role of exposure duration for altering responses, Pimephales promelas survival was compared following static (96 h) and pulse (1.5, 4, 8, and 15 h half-lives) exposures of CuSO4•5H2O. Copper concentrations sorbed by fry indicated a consequence of different exposures. Responses of P. promelas to static exposures resulted in 96 h LC50s of 166 µgCu/L (95% confidence interval [CI], 142-189 µgCu/L) as soluble copper and 162 µgCu/L (CI, 140-183 µgCu/L) as acid soluble copper. Relative to static 96 h LC50s, exposures with half-lives of 1.5, 4 and 8 h resulted in LC50s 10, 3 and 2 times greater, respectively, for responses measured 96 h after exposure initiation. Copper concentrations extracted from fry exposed for 1.5, 4 and 8 h half-lives were less than the static experiment. However, copper sorbed by fry in the 15 h half-life experiment was not different than the static experiment. The relationship between 96 h LC50 and 1/half-life was expressed using the equations y = 116 + 1360 × (R2 = 0.97) for soluble copper and y = 147 + 1620 × (R2 = 0.98) for acid soluble copper. Incorporation of exposure duration for predictions of P. promelas responses to copper pulse exposures increases prediction accuracy by an order of magnitude.

Influence of CuSO4 and chelated copper algaecide exposures on biodegradation of microcystin-LR.

Copper exposures from algaecide applications in aquatic systems are hypothesized to impede bacterial degradation of microcystin (MC), a cyanobacterial produced hepatotoxin. Despite regulatory implications of this hypothesis, limited data exist on influences of copper-exposures on MC-degrading bacteria and consequent MC-degradation. In this study, influences of copper-algaecide concentrations and formulations on bacterial composition and microcystin-LR (MCLR) degradation were investigated. Microcystis aeruginosa was exposed to four concentrations (0-5.0 mg Cu L-1) of three copper-algaecide formulations, and rates and extents of MCLR degradation were measured. In untreated controls and following exposures of 0.1, 0.5, and 1.0 mg Cu L-1, MCLR concentrations decreased at a rate of ∼41-53 µg MCLR/L d-1. Following exposure to 5.0 mg Cu L-1 MCLR degradation rates decreased an order of magnitude to ∼3-7 µg MCLR/L d-1. Bacterial diversity decreased following copper-exposures greater than 0.1 mg Cu L-1 for all formulations. Relative abundance of certain groups of MC-degrading bacteria identified in treatments increased with increasing copper concentration, suggesting they may be less sensitive to copper exposures than other, MCLR and non MC-degrading heterotrophic bacteria present in the assemblage. Results from this study revealed that copper concentration can influence degradation rates of MCLR, however this influence was not significant within copper concentrations currently registered for use (≤1.0 mg Cu L-1) of the tested algaecides. Copper formulation did not significantly alter degradation rates or bacterial composition. These data augment our understanding of the influences of copper algaecide-exposures on MCLR degradation, and can be used to inform more accurate risk evaluations and use of copper-algaecides for management of MCLR-producing cyanobacteria.

A risk-based approach for identifying constituents of concern in oil sands process-affected water from the Athabasca Oil Sands region.

Mining leases in the Athabasca Oil Sands (AOS) region produce large volumes of oil sands process-affected water (OSPW) containing constituents that limit beneficial uses and discharge into receiving systems. The aim of this research is to identify constituents of concern (COCs) in OSPW sourced from an active settling basin with the goal of providing a sound rational for developing mitigation strategies for using constructed treatment wetlands for COCs contained in OSPW. COCs were identified through several lines of evidence: 1) chemical and physical characterization of OSPW and comparisons with numeric water quality guidelines and toxicity endpoints, 2) measuring toxicity of OSPW using a taxonomic range of sentinel organisms (i.e. fish, aquatic invertebrates, and a macrophyte), 3) conducting process-based manipulations (PBMs) of OSPW to alter toxicity and inform treatment processes, and 4) discerning potential treatment pathways to mitigate ecological risks of OSPW based on identification of COCs, toxicological analyses, and PBM results. COCs identified in OSPW included organics (naphthenic acids [NAs], oil and grease [O/G]), metals/metalloids, and suspended solids. In terms of species sensitivities to undiluted OSPW, fish ≥ aquatic invertebrates > macrophytes. Bench-scale manipulations of the organic fractions of OSPW via PBMs (i.e. H2O2+UV254 and granular activated charcoal treatments) eliminated toxicity to Ceriodaphnia dubia (7-8 d), in terms of mortality and reproduction. Results from this study provide critical information to inform mitigation strategies using passive or semi-passive treatment processes (e.g., constructed treatment wetlands) to mitigate ecological risks of OSPW to aquatic organisms.

Comparative toxicity of sodium carbonate peroxyhydrate to freshwater organisms.

Sodium carbonate peroxyhydrate (SCP) is a granular algaecide containing H2O2 as an active ingredient to control growth of noxious algae. Measurements of sensitivities of target and non-target species to hydrogen peroxide are necessary for water resource managers to make informed decisions and minimize risks for non-target species when treating noxious algae. The objective of this study was to measure and compare responses among a target noxious alga (cyanobacterium Microcystis aeruginosa) and non-target organisms including a eukaryotic alga (chlorophyte Pseudokirchneriella subcapitata), microcrustacean (Ceriodaphnia dubia), benthic amphipod (Hyalella azteca), and fathead minnow (Pimephales promelas) to exposures of hydrogen peroxide as SCP. Hydrogen peroxide exposures were confirmed using the I3(-) method. SCP margins of safety for these organisms were compared with published toxicity data to provide context for other commonly used algaecides and herbicides (e.g. copper formulations, endothall, and diquat dibromide). Algal responses (cell density and chlorophyll a concentrations) and animal mortality were measured after 96h aqueous exposures to SCP in laboratory-formulated water to estimate EC50 and LC50 values, as well as potency slopes. Despite a shorter test duration, M. aeruginosa was more sensitive to hydrogen peroxide as SCP (96h EC50:0.9-1.0mgL(-)(1) H2O2) than the eukaryotic alga P. subcapitata (7-d EC50:5.2-9.2mgL(-1) H2O2), indicating potential for selective control of prokaryotic algae. For the three non-target animals evaluated, measured 96-h LC50 values ranged from 1.0 to 19.7mgL(-1) H2O2. C. dubia was the most sensitive species, and the least sensitive species was P. promelas, which is not likely to be affected by concentrations of hydrogen peroxide as SCP that would be used to control noxious algae (e.g. M. aeruginosa). Based on information from peer-reviewed literature, other algaecides could be similarly selective for cyanobacteria. Of the algaecides compared, SCP can selectively mitigate risks associated with noxious cyanobacterial growths (e.g. M. aeruginosa), with an enhanced margin of safety for non-target species (e.g. P. promelas).

Cellular and aqueous microcystin-LR following laboratory exposures of Microcystis aeruginosa to copper algaecides.

Microcystin release from algal cells influences use of copper-algaecides in water resources. Accurate data regarding relationships between copper-algaecide exposures and responses of microcystin-producing algae are needed to make informed management decisions. Responses of Microcystis aeruginosa were measured in terms of cellular microcystin-LR (MC-LR), aqueous MC-LR, and chlorophyll-a following exposure to CuSO4 and copper-ethanolamine. Comparisons were made between treated and untreated samples, and copper formulations. EC50s and slopes for M. aeruginosa responses to copper exposures were calculated. Algal responses followed a sigmoidal exposure-response relationship, and cellular MC-LR and chlorophyll-a were negatively related to copper concentrations. Aqueous MC-LR increased with copper concentrations, although the increase in aqueous MC-LR was not proportional to decreases in cellular MC-LR and chlorophyll-a. Cellular MC-LR and chlorophyll a declined at a greater rate than aqueous MC-LR increased. Total MC-LR was less than untreated controls following copper exposure. Differences were measured between copper formulations in terms of aqueous and total MC-LR concentrations at concentrations of 0.5 and 1.0 mg Cu L-1. Aqueous and total MC-LR were greater (10-20%) following exposure to CuSO4 compared to copper-ethanolamine one day following exposure. The positive relationship between copper concentration and aqueous MC-LR at 0.07-1.0 mg Cu L-1 demonstrates that lower copper concentrations were as effective as higher concentrations in controlling M. aeruginosa while decreasing the total amount of MC-LR, and minimizing the proportion of MC-LR released to the aqueous-phase. Results serve to support more accurate risk evaluations of MC-LR concentrations when M. aeruginosa is exposed to copper-algaecides and when it is untreated.

Responses of Lyngbya wollei to algaecide exposures and a risk characterization associated with their use.

To make informed decisions regarding management of noxious algal growths, water resource managers require information on responses of target and non-target species to algaecide exposures. Periodic treatments of Phycomycin®-SCP (sodium carbonate peroxyhydrate) followed by Algimycin®-PWF (gluconate and citrate chelated copper) to control Lyngbya wollei growths for ten years provided an opportunity for a risk evaluation of treated coves in Lay Lake, AL. Abiotic sediment characteristics (acid soluble copper concentrations, acid volatile sulfides, percent organic matter and cation exchange capacity) and survival of Hyalella azteca and Chironomus dilutus were measured in sediment samples from treated and untreated coves to assess the bioavailability of potential copper-residuals. In laboratory studies to seek a more effective approach for managing the growth of Lyngbya, six algaecide treatments consisting of combinations of copper-based algaecides (Cutrine®-Ultra, Clearigate® and Algimycin®- PWF), a hydrogen peroxide based algaecide (Phycomycin®-SCP) and an adjuvant (Cide-Kick II) were assessed for efficacy in controlling L. wollei sampled from Lay Lake. The most efficient algaecide treatment was determined based on post-treatment algal wet weight and visual observations of responses to exposures. To estimate the margin of safety for non-target organisms, Pimephales promelas was exposed to the most efficacious treatment and a treatment of Phycomycin®-SCP followed by Algimycin®-PWF. Results from sediment experiments demonstrated that there were no measureable copper residuals and no adverse effects on H. azteca and C. dilutus from sediments following ten years of copper-based algaecide treatments. Based on the laboratory results, a treatment of Phycomycin®-SCP at 10.1 mg H2O2/L followed by Cide-Kick II at 0.2 mg/L and Algimycin®- PWF at 0.26 mg Cu/L could control the growth of Lyngbya wollei from Lay Lake, AL and enhance the margin of safety for non-target species (e.g. P. promelas).

Evaluation of the utility of six measures for algal (Microcystis aeruginosa, Planktothrix agardhii and Pseudokirchneriella subcapitata) viability.

Standard algal toxicity tests are used to discern responses of algae to a variety of exposures including pesticides, personal care products and complex mixtures such as runoff and effluents. There are concerns regarding the accuracy, precision and utility of algal viability measures used as endpoints in algal toxicity test protocols. To definitively evaluate six algal viability measures, algae were heat-treated to produce known live:dead cell ratios. Cultures of two prokaryotic algae (Microcystis aeruginosa and Planktothrix agardhii) and a eukaryotic alga (Pseudokirchneriella subcapitata) were boiled for five minutes and mixed after cooling with untreated cultures to produce suspensions of 0%, 25%, 50%, 75% and 100% live algal cells. Optical microscopy was used to assess the viability of algae on a cell-by-cell basis by measuring cell density, uptake of a vital stain (neutral red) and exclusion of a mortal stain (erythrosin b). Aggregate measures of algal cell viability included chlorophyll a concentrations, pheophytin a concentrations and respiration (measured as 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium formazan absorbance (INT)). Cell densities, erythrosin b stained cells and chlorophyll a concentrations correlated with viable M. aeruginosa, P. agardhii and P. subcapitata cells (R(2)=0.97-0.78, 0.98-0.85 and 0.99-0.97 respectively). Pheophytin a concentrations and neutral red stained cells did not correlate with viable algae (R(2)=0.41-0.01 and 0.15-0.03 respectively). For INT formazan absorbance, 50%, 75% and 100% viable algae had greater variances and did not strongly correlate (R(2)=0.75-0.54). This result was likely confounded by respiration associated with resident bacteria. Three of the six methods provided accurate and precise information regarding the viability of both prokaryotic and eukaryotic algae. These methods also have a relatively low initial expense and can be used widely.