Understanding the risks of co-exposures in a changing world: a case study of dual monitoring of the biotoxin domoic acid and Vibrio spp. in Pacific oyster.

Assessing the co-occurrence of multiple health risk factors in coastal ecosystems is challenging due to the complexity of multi-factor interactions and limited availability of simultaneously collected data. Understanding co-occurrence is particularly important for risk factors that may be associated with, or occur in similar environmental conditions. In marine ecosystems, the co-occurrence of harmful algal bloom toxins and bacterial pathogens within the genus Vibrio may impact both ecosystem and human health. This study examined the co-occurrence of Vibrio spp. and domoic acid (DA) produced by the harmful algae Pseudo-nitzschia by (1) analyzing existing California Department of Public Health monitoring data for V. parahaemolyticus and DA in oysters; and (2) conducting a 1-year seasonal monitoring of these risk factors across two Southern California embayments. Existing public health monitoring efforts in the state were robust for individual risk factors; however, it was difficult to evaluate the co-occurrence of these risk factors in oysters due to low number of co-monitoring instances between 2015 and 2020. Seasonal co-monitoring of DA and Vibrio spp. (V. vulnificus or V. parahaemolyticus) at two embayments revealed the co-occurrence of these health risk factors in 35% of sampled oysters in most seasons. Interestingly, both the overall detection frequency and co-occurrence of these risk factors were considerably less frequent in water samples. These findings may in part suggest the slow depuration of Vibrio spp. and DA in oysters as residual levels may be retained. This study expanded our understanding of the simultaneous presence of DA and Vibrio spp. in bivalves and demonstrates the feasibility of co-monitoring different risk factors from the same sample. Individual programs monitoring for different risk factors from the same sample matrix may consider combining efforts to reduce cost, streamline the process, and better understand the prevalence of co-occurring health risk factors.

A cross-regional examination of patterns and environmental drivers of Pseudo-nitzschia harmful algal blooms along the California coast.

Pseudo-nitzschia species with the ability to produce the neurotoxin domoic acid (DA) are the main cause of harmful algal blooms (HABs) along the U.S. West Coast, with major impacts on ecosystems, fisheries, and human health. While most Pseudo-nitzschia (PN) HAB studies to date have focused on their characteristics at specific sites, few cross-regional comparisons exist, and mechanistic understanding of large-scale HAB drivers remains incomplete. To close these gaps, we compiled a nearly 20-year time series of in situ particulate DA and environmental observations to characterize similarities and differences in PN HAB drivers along the California coast. We focus on three DA hotspots with the greatest data density: Monterey Bay, the Santa Barbara Channel, and the San Pedro Channel. Coastwise, DA outbreaks are strongly correlated with upwelling, chlorophyll-a, and silicic acid limitation relative to other nutrients. Clear differences also exist across the three regions, with contrasting responses to climate regimes across a north to south gradient. In Monterey Bay, PN HAB frequency and intensity increase under relatively nutrient-poor conditions during anomalously low upwelling intensities. In contrast, in the Santa Barbara and San Pedro Channels, PN HABs are favored under cold, nitrogen-rich conditions during more intense upwelling. These emerging patterns provide insights on ecological drivers of PN HABs that are consistent across regions and support the development of predictive capabilities for DA outbreaks along the California coast and beyond.

Integrative monitoring strategy for marine and freshwater harmful algal blooms and toxins across the freshwater-to-marine continuum.

Many coastal states throughout the USA have observed negative effects in marine and estuarine environments caused by cyanotoxins produced in inland waterbodies that were transported downstream or produced in the estuaries. Estuaries and other downstream receiving waters now face the dual risk of impacts from harmful algal blooms (HABs) that occur in the coastal ocean as well as those originating in inland watersheds. Despite this risk, most HAB monitoring efforts do not account for hydrological connections in their monitoring strategies and designs. Monitoring efforts in California have revealed the persistent detection of cyanotoxins across the freshwater-to-marine continuum. These studies underscore the importance of inland waters as conduits for the transfer of cyanotoxins to the marine environment and highlight the importance of approaches that can monitor across hydrologically connected waterbodies. A HAB monitoring strategy is presented for the freshwater-to-marine continuum to inform HAB management and mitigation efforts and address the physical and hydrologic challenges encountered when monitoring in these systems. Three main recommendations are presented based on published studies, new datasets, and existing monitoring programs. First, HAB monitoring would benefit from coordinated and cohesive efforts across hydrologically interconnected waterbodies and across organizational and political boundaries and jurisdictions. Second, a combination of sampling modalities would provide the most effective monitoring for HAB toxin dynamics and transport across hydrologically connected waterbodies, from headwater sources to downstream receiving waterbodies. Third, routine monitoring is needed for toxin mixtures at the land-sea interface including algal toxins of marine origins as well as cyanotoxins that are sourced from inland freshwater or produced in estuaries. Case studies from California are presented to illustrate the implementation of these recommendations, but these recommendations can also be applied to inland states or regions where the downstream receiving waterbody is a freshwater lake, reservoir, or river. Integr Environ Assess Manag 2023;19:586-604. © 2022 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Reduced representation sequencing accurately quantifies relative abundance and reveals population-level variation in Pseudo-nitzschia spp.

Certain species within the genus Pseudo-nitzschia are able to produce the neurotoxin domoic acid (DA), which can cause illness in humans, mass-mortality of marine animals, and closure of commercial and recreational shellfisheries during toxic events. Understanding and forecasting blooms of these harmful species is a primary management goal. However, accurately predicting the onset and severity of bloom events remains difficult, in part because the underlying drivers of bloom formation have not been fully resolved. Furthermore, Pseudo-nitzschia species often co-occur, and recent work suggests that the genetic composition of a Pseudo-nitzschia bloom may be a better predictor of toxicity than prevailing environmental conditions. We developed a novel next-generation sequencing assay using restriction site-associated DNA (2b-RAD) genotyping and applied it to mock Pseudo-nitzschia communities generated by mixing cultures of different species in known abundances. On average, 94% of the variance in observed species abundance was explained by the expected abundance. In addition, the false positive rate was low (0.45% on average) and unrelated to read depth, and false negatives were never observed. Application of this method to environmental DNA samples collected during natural Pseudo-nitzschia spp. bloom events in Southern California revealed that increases in DA were associated with increases in the relative abundance of P. australis. Although the absolute correlation across time-points was weak, an independent species fingerprinting assay (Automated Ribosomal Intergenic Spacer Analysis) supported this and identified other potentially toxic species. Finally, we assessed population-level genomic variation by mining SNPs from the environmental 2bRAD dataset. Consistent shifts in allele frequencies in P. pungens and P. subpacifica were detected between high and low DA years, suggesting that different intraspecific variants may be associated with prevailing environmental conditions or the presence of DA. Taken together, this method presents a potentially cost-effective and high-throughput approach for studies aiming to evaluate both population and species dynamics in mixed samples.

Daily dynamics of contrasting spring algal blooms in Santa Monica Bay (central Southern California Bight).

Protistan algae (phytoplankton) dominate coastal upwelling ecosystems where they form massive blooms that support the world's most important fisheries and constitute an important sink for atmospheric CO2 . Bloom initiation is well understood, but the biotic and abiotic forces that shape short-term dynamics in community composition are still poorly characterized. Here, high-frequency (daily) changes in relative abundance dynamics of the metabolically active protistan community were followed via expressed 18S V4 rRNA genes (RNA) throughout two algal blooms during the spring of 2018 and 2019 in Santa Monica Bay (central Southern California Bight). A diatom bloom formed after wind-driven, nutrient upwelling events in both years, but different taxa dominated each year. Whereas diatoms bloomed following elevated nutrients and declined after depletion each year, a massive dinoflagellate bloom manifested under relatively low inorganic nitrogen conditions following diatom bloom senescence in 2019 but not 2018. Network analysis revealed associations between diatoms and cercozoan putative parasitic taxa and syndinean parasites during 2019 that may have influenced the demise of the diatoms, and the transition to a dinoflagellate-dominated bloom.

Development and field evaluation of the organic-diffusive gradients in thin-films (o-DGT) passive water sampler for microcystins.

The presence of microcystins (MCs) in waterbodies requires a simple and reliable monitoring technique to characterize better their spatiotemporal distribution and ecological risks. An organic-diffusive gradients in thin films (o-DGT) passive sampler based on polyacrylamide diffusive gel and hydrophilic-lipophilic balance (HLB) binding gel was developed for MCs in water. The mass accumulation of three MCs (MC-LR, -RR, and -YR) was linear over 10 days (R2 ≥ 0.98). Sampling rates (2.68-3.22 mL d-1) and diffusion coefficients (0.90-1.08 × 10-6 cm2 s-1) of three MCs were obtained at 20 °C. Two different passive samplers, o-DGT and the Solid Phase Adsorption Toxin Tracking device (SPATT), were co-deployed to estimate MC levels at three lakes in California, USA. Measured total MC concentrations were up to 10.9 µg L-1, with MC-LR the primary variant at a measured maximum concentration of 2.74 µg L-1. Time-weighted average MC concentrations by o-DGT were lower than grab water samples, probably because grab sampling measures both dissolved and particulate phases (i.e., MCs in cyanobacteria). Passive water samplers by design can only measure dissolved-phase MCs, which are considerably less during the cyanobacteria-laden periods observed. Both o-DGT and grab samples gave comparable results for three MC variants at low levels of MCs, e.g., <0.1 µg L-1. o-DGT showed a higher correlation with grab sampling than SPATT did. This study demonstrates that o-DGT can be effectively used for monitoring and evaluation of dissolved MCs in waters.

Persistent domoic acid in marine sediments and benthic infauna along the coast of Southern California.

Blooms of the diatom genus Pseudo-nitzschia occur annually in the Southern California Bight (SCB), and domoic acid (DA) associated with these events can contaminate fisheries, presenting both human and wildlife health risks. Recent studies have suggested that marine sediments may act as a reservoir for DA, extending the risk of food web contamination long after water column blooms have ended. In this study, we conducted a regional assessment of the extent and magnitude of DA in the benthic environment, and monthly observations of sediments and benthic infauna at multiple stations over a 16-month period. DA was widespread in continental shelf sediments of the SCB. The toxin was detected in 54% of all shelf habitats sampled. Detectable concentrations ranged from 0.11 ng/g to 1.36 ng/g. DA was consistently detected in benthic infauna tissues over the monthly timeseries, while the DA concentrations in sediments during the same period were commonly below detection or at low concentrations. The presence of DA in the benthic environment did not always have an apparent water column source, raising the possibility of lateral transport, retention/preservation in sediments or undetected blooms in subsurface waters. In most cases, DA was detected in tissues but not in the co-located surface sediments. Coarse taxonomic sorting of the infauna revealed that the accumulation of DA varied among taxa. We observed that DA was widespread among lower trophic level organisms in this study, potentially acting as a persistent source of DA to higher trophic levels in the benthos.

Synthesis of ecotoxicological studies on cyanotoxins in freshwater habitats - Evaluating the basis for developing thresholds protective of aquatic life in the United States.

In recent decades, cyanobacteria harmful algal blooms (cyanoHABs) have increased in magnitude, frequency, and duration in freshwater ecosystems. CyanoHABs can impact water quality by the production of potent toxins known as cyanotoxins. Environmental exposure to cyanotoxins has been associated with severe illnesses in humans, domestic animals, and wildlife. However, the effects of sustained exposure to cyanotoxins on aquatic life are poorly understood. In this study, over 150 peer-reviewed articles were critically evaluated to better understand the ecotoxicity of 5 cyanotoxin classes (microcystins, cylindrospermopsin, anatoxin-a, saxitoxins, nodularin) on fish, amphibians, aquatic invertebrates, and birds exclusively feeding in freshwater habitats. The systemic review demonstrated that microcystins, and more specifically microcystin-LR, were the most studied cyanotoxins. Ecotoxicological investigations were typically conducted using a fish or aquatic invertebrate model, with mortality, bioaccumulation, and biochemical responses as the most frequently measured endpoints. After excluding the studies that did not meet our acceptability criteria, remaining studies were examined to identify the no-observed and lowest observed effect concentrations (NOEC and LOEC) for microcystins; the limited amount of data for other cyanotoxins did not allow for analysis. The published ecotoxicity data suggests that the U.S. EPA recreational water quality criteria for microcystin (8 µg/L) may be protective of acute toxicity in aquatic organisms but does not appear to protect against chronic toxicity. Individual U.S. states have developed more stringent recreational health-based thresholds, such as 0.8 µg/L in California. Comparisons of this threshold to the chronic NOEC and LOEC data indicate that more restrictive microcystins thresholds may be required to be protective of aquatic life. Additional research is needed to evaluate the sublethal effects of a wider array of microcystin congeners and other cyanotoxins on organisms relevant to U.S. watersheds to better support nationwide thresholds protective of aquatic life.

The tide turns: Episodic and localized cross-contamination of a California coastline with cyanotoxins.

The contamination of coastal ecosystems from a variety of toxins of marine algal origin is a common and well-documented situation along the coasts of the United States and globally. The occurrence of toxins originating from cyanobacteria along marine coastlines is much less studied, and little information exists on whether toxins from marine and freshwater sources co-occur regularly. The current study focused on the discharge of cyanotoxins from a coastal lagoon (Santa Clara River Estuary) as a consequence of an extreme tide event (King Tides; December 3-5, 2017) resulting in a breach of the berm separating the lagoon from the ocean. Monthly monitoring in the lagoon throughout 2017 documented more than a dozen co-occurring cyanobacterial genera, as well as multiple algal and cyanobacterial toxins. Biotoxin monitoring before and following the King Tide event using Solid Phase Adsorption Toxin Tracking (SPATT) in the lagoon and along the coast revealed the co-occurrence of microcystins, anatoxin, domoic acid, and other toxins on multiple dates and locations. Domoic acid was ubiquitously present in SPATT deployed in the lagoon and along the coast. Microcystins were also commonly detected in both locations, although the beach berm retained the lagoonal water for much of the year. Mussels collected along the coast contained microcystins in approximately half the samples, particularly following the King Tide event. Anatoxin was observed in SPATT only in late December, following the breach of the berm. Our findings indicate both episodic and persistent occurrence of both cyanotoxins and marine toxins may commonly contaminate coastlines in proximity to cyanobacteria-laden creeks and lagoons.

Co-occurring dissolved algal toxins observed at multiple coastal sites in southern California via solid phase adsorption toxin tracking.

Algal toxins (domoic acid, saxitoxin, okadaic acid) were monitored at seven locations off southern California using Solid Phase Adsorption Toxin Tracking. At least two types of toxins were found at all locations, with co-occurrence of two and three toxins in 12% and 10% of samples, respectively. This study expands our limited understanding of the simultaneous presence of multiple algal toxins along the coast and raises questions regarding the potential health ramifications of such co-occurrences.

A decade and a half of Pseudo-nitzschia spp. and domoic acid along the coast of southern California.

Blooms of the marine diatom genus Pseudo-nitzschia that produce the neurotoxin domoic acid have been documented with regularity along the coast of southern California since 2003, with the occurrence of the toxin in shellfish tissue predating information on domoic acid in the particulate fraction in this region. Domoic acid concentrations in the phytoplankton inhabiting waters off southern California during 2003, 2006, 2007, 2011 and 2017 were comparable to some of the highest values that have been recorded in the literature. Blooms of Pseudo-nitzschia have exhibited strong seasonality, with toxin appearing predominantly in the spring. Year-to-year variability of particulate toxin has been considerable, and observations during 2003, 2006, 2007, 2011 and again in 2017 linked domoic acid in the diets of marine mammals and seabirds to mass mortality events among these animals. This work reviews information collected during the past 15 years documenting the phenology and magnitude of Pseudo-nitzschia abundances and domoic acid within the Southern California Bight. The general oceanographic factors leading to blooms of Pseudo-nitzschia and outbreaks of domoic acid in this region are clear, but subtle factors controlling spatial and interannual variability in bloom magnitude and toxin production remain elusive.