Synchronic distribution of the dinoflagellate Protoceratium reticulatum and yessotoxins in a high stratified fjord system: Tidal or light modulation?

Protoceratium reticulatum is the main yessotoxin-producer along the Chilean coast. Thus far, the yessotoxin levels recorded in this region have not posed a serious threat to human health. However, a bloom of P. reticulatum during the austral summer of 2022 caused the first ban of shellfish collection, due to the high toxin levels. A bloom of P. reticulatum during the austral summer of 2020 allowed an evaluation of the fine-scale distribution of the dinoflagellate during a tidal cycle. High-resolution measurements of biophysical properties were carried out in mid-summer (February 18-19) at a fixed sampling station in Puyuhuapi Fjord, Chilean Patagonia, as part of an intensive 24-h biophysical experiment to monitor the circadian distributions of P. reticulatum vegetative cells and yessotoxins. High P. reticulatum cell densities (>20 × 103 cells L-1) were found in association with a warmer (14.5-15 °C) and estuarine (23.5-24.5 g kg-1) sub-surface water layer (6-8 m). P. reticulatum cell numbers and yessotoxins followed a synchronic distribution pattern consistent with the excursions of the pycnocline. Nevertheless, the surface aggregation of the cells was modulated by the light cycle, suggesting daily vertical migration. The yessotoxin content per P. reticulatum cell ranged from 9.4 to 52.2 pg. This study demonstrates both the value of fine-scale resolution measurements of biophysical properties in a highly stratified system and the potential ecosystem impact of P. reticulatum strains producing high levels of yessotoxins.

An Unprecedented Bloom of Oceanic Dinoflagellates (Karenia spp.) Inside a Fjord within a Highly Dynamic Multifrontal Ecosystem in Chilean Patagonia.

At the end of summer 2020, a moderate (~105 cells L-1) bloom of potential fish-killing Karenia spp. was detected in samples from a 24 h study focused on Dinophysis spp. in the outer reaches of the Pitipalena-Añihue Marine Protected Area. Previous Karenia events with devastating effects on caged salmon and the wild fauna of Chilean Patagonia had been restricted to offshore waters, eventually reaching the southern coasts of Chiloé Island through the channel connecting the Chiloé Inland Sea to the Pacific Ocean. This event occurred at the onset of the COVID-19 lockdown when monitoring activities were slackened. A few salmon mortalities were related to other fish-killing species (e.g., Margalefidinium polykrikoides). As in the major Karenia event in 1999, the austral summer of 2020 was characterised by negative anomalies in rainfall and river outflow and a severe drought in March. Karenia spp. appeared to have been advected in a warm (14-15 °C) surface layer of estuarine saline water (S > 21). A lack of daily vertical migration patterns and cells dispersed through the whole water column suggested a declining population. Satellite images confirmed the decline, but gave evidence of dynamic multifrontal patterns of temperature and chl a distribution. A conceptual circulation model is proposed to explain the hypothetical retention of the Karenia bloom by a coastally generated eddy coupled with the semidiurnal tides at the mouth of Pitipalena Fjord. Thermal fronts generated by (topographically induced) upwelling around the Tic Toc Seamount are proposed as hot spots for the accumulation of swimming dinoflagellates in summer in the southern Chiloé Inland Sea. The results here provide helpful information on the environmental conditions and water column structure favouring Karenia occurrence. Thermohaline properties in the surface layer in summer can be used to develop a risk index (positive if the EFW layer is thin or absent).

Small and patchy is enough: An example about how toxic HAB events can spread through low resting cyst loads.

The frequency of harmful algal blooms (HABs) has increased over the last two decades, a phenomenon enhanced by global climate change. However, the effects of climate change will not be distributed equally, and Chile has emerged as one important, vulnerable area. The Chilean Patagonian region (41‒56°S) hosts two marine ecoregions that support robust blue economies via wild fisheries, aquaculture, and tourism. However, the harmful algal bloom-forming dinoflagellate Alexandrium catenella, a causative agent of paralytic shellfish poisoning outbreaks, threatens the viability of blue industries in this region and others worldwide. Despite the proliferation of A. catenella blooms over the last few decades, the role of sedimentary resting cysts in the recurrence of harmful algal blooms and the species' northward expansion across Chilean Patagonia is not well understood. As a resting cyst-producing species, the sediment-cyst dynamics of A. catenella likely contribute to the geographical expansion and bloom recurrence of this species. For this purpose, we analyzed a decade of A. catenella surface sediment cyst records across the two ecoregions of the Chilean Patagonian System that were further stratified into five subregions based on water temperature, salinity, dissolved oxygen, and nutrient characteristics. We also analyzed spatio-temporal cyst dynamics in a pre-, during-, and post-bloom scenario of the Chiloense ecoregion (more northern) of the Magellanic province. Our results indicated highly variable A. catenella resting cyst abundances, with a maximum of 221 cysts cm-3 recorded in 2002 after an intense bloom. Generalized linear mixed models and linear mixed models found that sampling season, subregion, and Total Organic Matter (%) explained resting cyst presence and density. The results also demonstrated the presence of A. catenella cysts in northern subregions, evidencing the northward geographical expansion observed during the last few decades. The risks of A. catenella bloom recurrence from small, patchy resting cyst distributions across broad geographical areas and under changing environmental conditions are discussed.

From lipophilic to hydrophilic toxin producers: Phytoplankton succession driven by an atmospheric river in western Patagonia.

Phytoplankton succession is related to hydroclimatic conditions. In this study we provide the first description of a toxic phytoplankton succession in the Patagonian Fjord System. The shift was modulated by atmospheric-oceanographic forcing and consisted of the replacement of the marine dinoflagellate Dinophysis acuta in a highly stratified water column during austral summer by the diatom Pseudo-nitzschia calliantha in a mixed water column during late summer and early autumn. This transition, accompanied by a change in the biotoxin profiles (from lipophilic dinophysis toxins to hydrophilic domoic acid), was induced by the arrival of an intense atmospheric river. The winds in Magdalena Sound may have been further amplified, due to its west-east orientation and its location within a tall, narrow mountain canyon. This work also documents the first known appearance of toxic P. calliantha in Northern Patagonian. The potential impacts of the biotoxins of this species on higher trophic levels are discussed.

Dinophysis acuminata or Dinophysis acuta: What Makes the Difference in Highly Stratified Fjords?

Dinophysis acuminata and D. acuta, which follows it seasonally, are the main producers of lipophilic toxins in temperate coastal waters, including Southern Chile. Strains of the two species differ in their toxin profiles and impacts on shellfish resources. D. acuta is considered the major cause of diarrhetic shellfish poisoning (DSP) outbreaks in Southern Chile, but there is uncertainty about the toxicity of D. acuminata, and little information on microscale oceanographic conditions promoting their blooms. During the austral summer of 2020, intensive sampling was carried out in two northern Patagonian fjords, Puyuhuapi (PUY) and Pitipalena (PIT), sharing D. acuminata dominance and D. acuta near detection levels. Dinophysistoxin 1 (DTX 1) and pectenotoxin 2 (PTX 2) were present in all net tow samples but OA was not detected. Although differing in hydrodynamics and sampling dates, D. acuminata shared behavioural traits in the two fjords: cell maxima (>103 cells L-1) in the interface (S ~ 21) between the estuarine freshwater (EFW)) and saline water (ESW) layers; and phased-cell division (µ = 0.3-0.4 d-1) peaking after dawn, and abundance of ciliate prey. Niche analysis (Outlying Mean Index, OMI) of D. acuta with a high marginality and much lower tolerance than D. acuminata indicated an unfavourable physical environment for D. acuta (bloom failure). Comparison of toxin profiles and Dinophysis niches in three contrasting years in PUY-2020 (D. acuminata bloom), 2018 (exceptional bloom of D. acuta), and 2019 (bloom co-occurrence of the two species)-shed light on the vertical gradients which promote each species. The presence of FW (S < 11) and thermal inversion may be used to provide short-term forecasts of no risk of D. acuta blooms and OA occurrence, but D. acuminata associated with DTX 1 pose a risk of DSP events in North Patagonian fjords.

The impact of local and climate change drivers on the formation, dynamics, and potential recurrence of a massive fish-killing microalgal bloom in Patagonian fjord.

Harmful algal blooms (HABs) in southern Chile are a serious threat to public health, tourism, artisanal fisheries, and aquaculture in this region. Ichthyotoxic HAB species have recently become a major annual threat to the Chilean salmon farming industry, due to their severe economic impacts. In early austral autumn 2021, an intense bloom of the raphidophyte Heterosigma akashiwo was detected in Comau Fjord, Chilean Patagonia, resulting in a high mortality of farmed salmon (nearly 6000 tons of biomass) within 15 days. H. akashiwo cells were first detected at the head of the fjord on March 16, 2021 (up to 478 cells mL-1). On March 31, the cell density at the surface had reached a maximum of 2 × 105 cells mL-1, with intense brown spots visible on the water surface. Strong and persistent high-pressure anomalies over the southern tip of South America, consistent with the positive phase of the Southern Annular Mode (SAM), resulted in extremely dry conditions, high solar radiation, and strong southerly winds. A coupling of these features with the high water retention times inside the fjord can explain the spatial-temporal dynamics of this bloom event. Other factors, such as the internal local physical uplift process (favored by the north-to-south orientation of the fjord), salt-fingering events, and the uplift of subantarctic deep-water renewal, likely resulted in the injection of nutrients into the euphotic layer, which in turn could have promoted cell growth and thus high microalgal cell densities, such as reached by the bloom.

Modelling the Spatial and Temporal Dynamics of Paralytic Shellfish Toxins (PST) at Different Scales: Implications for Research and Management.

Harmful algal blooms, in particular recurrent blooms of the dinoflagellate Alexandrium catenella, associated with paralytic shellfish poisoning (PSP), frequently limit commercial shellfish harvests, resulting in serious socio-economic consequences. Although the PSP-inducing species that threaten the most vulnerable commercial species of shellfish are very patchy and spatially heterogeneous in their distribution, the spatial and temporal scales of their effects have largely been ignored in monitoring programs and by researchers. In this study, we examined the spatial and temporal dynamics of PSP toxicity in the clam (Ameghinomya antiqua) in two fishing grounds in southern Chile (Ovalada Island and Low Bay). During the summer of 2009, both were affected by an intense toxic bloom of A. catenella (up to 1.1 × 106 cells L-1). Generalized linear models were used to assess the potential influence of different environmental variables on the field detoxification rates of PSP toxins over a period of 12 months. This was achieved using a four parameter exponential decay model to fit and compare field detoxification rates per sampling site. The results show differences in the spatial variability and temporal dynamics of PSP toxicity, given that greater toxicities (+10-fold) and faster detoxification (20% faster) are observed at the Ovalada Island site, the less oceanic zone, and where higher amounts of clam are annually produced. Our observations support the relevance of considering different spatial and temporal scales to obtain more accurate assessments of PSP accumulation and detoxification dynamics and to improve the efficacy of fisheries management after toxic events.

Continental shelf off northern Chilean Patagonia: A potential risk zone for the onset of Alexandrium catenella toxic bloom?

Harmful Algal Blooms (HAB) pose a severe socio-economic problem worldwide. The dinoflagellate species Alexandrium catenella produces potent neurotoxins called saxitoxins (STXs) and its blooms are associated with the human intoxication named Paralytic Shellfish Poisoning (PSP). Knowing where and how these blooms originate is crucial to predict blooms. Most studies in the Chilean Patagonia, were focused on coastal areas, considering that blooms from the adjacent oceanic region are almost non-existent. Using a combination of field studies and modelling approaches, we first evaluated the role of the continental shelf off northern Chilean Patagonia as a source of A. catenella resting cysts, which may act as inoculum for their toxic coastal blooms. This area is characterized by a seasonal upwelling system with positive Ekman pumping during spring-summer, and by the presence of six major submarine canyons. We found out that these submarine canyons increase the vertical advection of bottom waters, and thus, significantly enhance the process of coastal upwelling. This is a previously unreported factor, among those involved in bloom initiation. This finding put this offshore area at high risk of resuspension of resting cysts of A. catenella. Here, we discuss in detail the physical processes promoting this resuspension.

Oceanography time series reveals annual asynchrony input between oceanic and estuarine waters in Patagonian fjords.

The postglacial Patagonian fjord system along the west coast of southern South America is one of the largest stretches of the southern hemisphere (SH) fjord belt, influenced by the SH westerly wind belt and continental freshwater input. This study reports a 3-year monthly time series (2017-2020) of physical and biogeochemical parameters obtained from the Reloncaví Marine Observatory (OMARE, Spanish acronym) at the northernmost embayment and fjord system of Patagonia. The main objective of this work was to understand the land-atmosphere-ocean interactions and to identify the mechanisms that modulate the density of phytoplankton. A key finding of this study was the seasonally varying asynchronous input of oceanic and estuarine water. Surface lower salinity and warmer estuarine water arrived in late winter to summer, contributing to water column stability, followed by subsurface higher salinity and less warmer oceanic water during fall-winter. In late winter 2019, an interannual change above the picnocline due to the record-high polarity of the Indian Ocean Dipole inhibited water column stability. The biogeochemical parameters (NO3-, NO2-, PO43-, Si(OH)4, pH, and dissolved oxygen) responded to the surface annual salinity variations, and oceanic water mass contributed greatly to the subsurface inorganic nutrient input. The water column N/P ratio indicated that no eutrophication occurred, even under intense aquaculture activity, likely because of the high ventilation dynamics of the Reloncaví Sound. Finally, a shift in phytoplankton composition, characterized by surface chlorophyll-a maxima in late winter and deepening of spring-summer blooms related to the physicochemical conditions of the water column, was observed. Our results support the ecosystem services provided by local oceanography processes in the north Patagonian fjords. Here, the anthropogenic impact caused by economic activities could be, in part, chemically reduced by the annual ventilation cycle mediated by the exchange of oceanic water masses into Patagonian fjords.

Niche differentiation of Dinophysis acuta and D. acuminata in a stratified fjord.

Dinophysis acuta and D. acuminata are associated with lipophilic toxins in Southern Chile. Blooms of the two species coincided during summer 2019 in a highly stratified fjord system (Puyuhuapi, Chilean Patagonia). High vertical resolution measurements of physical parameters were carried out during 48 h sampling to i) explore physiological status (e.g., division rates, toxin content) and ii) illustrate the fine scale distribution of D. acuta and D. acuminata populations with a focus on water column structure and co-occurring plastid-bearing ciliates. The species-specific resources and regulators defining the realized niches (sensu Hutchinson) of the two species were identified. Differences in vertical distribution, daily vertical migration and in situ division rates (with record values, 0.76 d-1, in D. acuta), in response to the environmental conditions and potential prey availability, revealed their niche differences. The Outlying Mean Index (OMI) analysis showed that the realized niche of D. acuta (cell maximum 7 × 103 cells L-1 within the pycnocline) was characterized by sub-surface estuarine waters (salinity 23 - 25), lower values of turbulence and PAR, and a narrow niche breath. In contrast, the realized niche of D. acuminata (cell maximum 6.8 × 103 cells L-1 just above the pycnocline) was characterized by fresher (salinity 17 - 20) outflowing surface waters, with higher turbulence and light intensity and a wider niche breadth. Results from OMI and PERMANOVA analyses of co-occurring microplanktonic ciliates were compatible with the hypothesis of species such as those from genera Pseudotontonia and Strombidium constituting an alternative ciliate prey to Mesodinium. The D. acuta cell maximum was associated with DSP (OA and DTX-1) toxins and pectenotoxins; that of D. acuminata only with pectenotoxins. Results presented here contribute to a better understanding of the environmental drivers of species-specific blooms of Dinophysis and management of their distinct effects in Southern Chile.

Multiscale physical background to an exceptional harmful algal bloom of Dinophysis acuta in a fjord system.

Dinophysis acuta produces diarrhetic shellfish poisoning (DSP) toxins and pectenotoxins (PTX). It blooms in thermally-stratified shelf waters in late summer in temperate to cold temperate latitudes. Despite its major contribution to shellfish harvesting bans, little effort has been devoted to study its population dynamics in Chilean Patagonia. In 2017-2018, mesoscale distribution of harmful algal species (75 monitoring stations) revealed the initiation (late spring) and seasonal growth of a dense D. acuta population in the Aysén region, with maximal values at Puyuhuapi Fjord (PF). Vertical phytoplankton distribution and fine-resolution measurements of physical parameters along a 25-km transect in February 16th identified a 15-km (horizontal extension) subsurface thin layer of D. acuta from 4 to 8 m depth. This layer, disrupted at the confluence of PF with the Magdalena Sound, peaked at the top of the pycnocline (6 m, 15.9 °C, 23.4 psu) where static stability was maximal. By February 22nd, it deepened (8 m, 15.5 °C; 23.62 psu) following the excursions of the pycnocline and reached the highest density ever recorded (664 × 103 cells L-1) for this species. Dinophysis acuta was the dominant Dinophysis species in all microplankton net-tows/bottle samples; they all contained DSP toxins (OA, DTX-1) and PTX-2. Modeled flushing rates showed that Puyuhuapi, the only fjord in the area with 2 connections with the open sea, had the highest water residence time. Long term climate variability in the Southern hemisphere showed the effects of a Southern Annular Mode (SAM) in positive mode (+1.1 hPa) overwhelming a moderate La Niña. These effects included positive spring precipitation anomalies with enhanced salinity gradients and summer drought with positive anomalies in air (+1 °C) and sea surface (+2 °C) temperature. Locally, persistent thermal stratification in PF seemed to provide an optimal physical habitat for initiation and bloom development of D. acuta. Thus, in summer 2018, a favourable combination of meteorological and hydrographic processes of multiple scales created conditions that promoted the development of a widespread bloom of D. acuta with its epicentre at the head of Puyuhuapi fjord.

Dinophysis Ehrenberg (Dinophyceae) in Southern Chile harbours red cryptophyte plastids from Rhodomonas/Storeatula clade.

Photosynthetic species of the dinoflagellate genus Dinophysis are known to retain temporary cryptophyte plastids of the Teleaulax/Plagioselmis/Geminigera clade after feeding the ciliate Mesodinium rubrum. In the present study, partial plastid 23S rDNA sequences were retrieved in Southern Chilean waters from oceanic (Los Lagos region), and fjord systems (Aysén region), in single cells of Dinophysis and accompanying organisms (the heliozoan Actinophrys cf. sol and tintinnid ciliates), identified by means of morphological discrimination under the light microscope. All plastid 23S rDNA sequences (n = 23) from Dinophysis spp. (Dinophysis acuta, D. caudata, D. tripos and D. subcircularis) belonged to cryptophytes from clade V (Rhinomonas, Rhodomonas and Storeatula), although they could not be identified at genus level. Moreover, five plastid sequences obtained from heliozoans (Actinophryida, tentatively identified as Actinophrys cf. sol), and tintinnid ciliates, grouped together with those cryptophyte sequences. In contrast, two additional sequences from tintinnids belonged to other taxa (chlorophytes and cyanobacteria). Overall, the present study represents the first time that red cryptophyte plastids outside of the Teleaulax/Plagioselmis/Geminigera clade dominate in wild photosynthetic Dinophysis spp. These findings suggest that either Dinophysis spp. are able to feed on other ciliate prey than Mesodinium and/or that cryptophyte plastids from clade V prevail in members of the M. rubrum species complex in the studied area.

Diel vertical migration into anoxic and high-pCO2 waters: acoustic and net-based krill observations in the Humboldt Current.

A select group of marine organisms can enter the Oxygen Minimum Zones (OMZs) and even anoxic waters, while performing diel vertical migration (DVM). DVM of the euphausiid Euphausia eximia off northern Chile in the spring of 2015 was documented based on acoustic measurements using an echo sounder along with net samplings. Dissolved oxygen (DO) concentrations were obtained using a vertical profiler, and water samples were collected to obtain in situ nitrite (NO2-) concentrations as well as pHT, total alkalinity (AT), and therefore carbon dioxide partial pressure (pCO2) was estimated. Krill were found to migrate up to the surface (0-50 m) during the night and returned to ca. 200-300 m depth during the day, spending between 11 and 14 h at these layers. At the surface, DO and NO2- concentrations were 208 and 0.14 µM respectively, while pHT was 8.04 and 405 µatm pCO2. In contrast, at the deeper layers (200-300 m), DO and NO2- were < 3 and 6.3 µM respectively, with pHT 7.53 and 1490 µatm pCO2. The pHT and high pCO2 values at depths represent the conditions predicted for open ocean waters in a worst-case global warming scenario by 2150. The acoustic scatter suggested that > 60% of the krill swarms enter the OMZ and anoxic waters during the daytime. These frequent migrations suggest that krill can tolerate such extreme conditions associated with anoxic and high-pCO2 waters. The inferences drawn from the observation of these migrations might have strong implications for the current oceanic carbon pump models, highlighting the need for understanding the molecular and physiological adaptations allowing these migrations.

Alexandrium catenella cyst accumulation by passive and active dispersal agents: Implications for the potential spreading risk in Chilean Patagonian fjords.

The dinoflagellate Alexandrium catenella is responsible for paralytic shellfish poisoning and negative socioeconomic impacts on the fishing industry and aquaculture. In Chilean Patagonia, the reasons underlying the significant increase in the geographical extension (from south to north) of A. catenella blooms during the last five decades are not well understood. To assess the potential spreading risk of A. catenella during an intense austral summer bloom, we conducted an in situ experiment in a "hotspot" of this dinoflagellate in southern Chile. The objective was to assess the accumulation of A. catenella resting cysts in passive (fishing nets) and active (mussels) dispersal agents during the phase of bloom decline. Large numbers of resting cysts were detected in fishing nets (maximum of 5334 cysts net-1 per month) at 5 m depth and in mussels (maximum of 16 cysts g-1 of digestive gland) near Vergara Island. The potential of these vectors to serve as inoculum sources and the implications of our findings for A. catenella population dynamics are discussed.

Cell Cycle, Division Rate, and Feeding of the Heterotroph Phalacroma rotundatum in a Chilean Fjord.

Phalacroma rotundatum is a rare cosmopolitan heterotrophic dinoflagellate. This species, included in the IOC-UNESCO Taxonomic Reference List of Harmful Microalgae, may be a diarrhetic shellfish poisoning (DSP) toxin vector, but little is known about its ecophysiology and behavior. A vertical net haul collected during the austral summer of 2018 in Reloncaví Sound (Chilean Patagonia) revealed an unusually abundant population of P. rotundatum and prompted intensive 24 h sampling on 16-17 January to study the cell cycle and feeding behavior of this species. Hydrographic measurements from a buoy revealed the local characteristic estuarine circulation, with a brackish surface layer (salinity 26-28) separated from saltier, colder bottom waters by a pycnocline at a depth modulated by the tidal regime. A high proportion of P. rotundatum cells were packed with digestive vacuoles (peak of 70% at 14:00), and phased cell division (µ = 0.46 d-1) occurred 3 h after sunrise. The division time (TD) was 2 h. This is the first cell cycle study of P. rotundatum. The results here disagree with those of previous field studies that considered asynchronous division in some Dinophysis species to be related to heterotrophic feeding. They also question the very specific prey requirements, Tiarina fusus, reported for P. rotundatum in northern Europe.