Are the mitochondrial cox1 and cob genes suitable markers for species of Dinophysis Ehrenberg?

The identification of Dinophysis species with similar morphology but different toxic (Diarrhetic Shellfish Poisoning, DSP) potential is a crucial task in harmful algae monitoring programmes. The taxonomic assignment of Dinophysis species using molecular markers is a difficult task due to extremely low interspecific variability within their nuclear ribosomal genes and intergenic regions. Mitochondrial cox1 gene has been proposed as a better specific marker for Dinophysis species based on its higher resolution for two morphologically related species (Dinophysis acuminata and Dinophysis ovum) of the “Dinophysis acuminata complex”. In this study, the potential of two mitochondrial genes (mt cox1 and cob) to discriminate among six Dinophysis species (field isolates and cultures) associated with DSP events was explored. Neither mt cox1 nor cob genes provided enough resolution for all species of Dinophysis. The cob gene showed very poor resolution and grouped all Dinophysis spp. in a common clade. In contrast, the cox1 phylogeny distinguished 5 clades in the Dinophysiales – the “acuminata complex”, the “caudata group”, “acuta + norvegica” and Phalacromaspp. However, within the “D. acuminata complex” mtcox1 is so far the unique marker that differentiates D. acuminata from other species: isolates of D. ovum and Dinophysis sacculus had almost identical sequences (only four mismatches), but they were well separated from D. acuminata. D. acuminata and Dinophysis skagii (considered a life cycle stage of the former) showed identical cox1 sequences. Probes towards this gene can be useful in Mediterranean and Western Iberia sites where the co-occurrence of close morphotypes of D. acuminata and D. sacculus pose a problem for monitoring analyses. This is the first report on cultures of D. sacculus and its phylogenetic relation with other species of the D. acuminata complex.     

Scales characterising a high density thin layer of Dinophysis acuta Ehrenberg and its transport within a coastal jet

An investigation into the distribution of Dinophysis spp. in coastal waters off the south coast of Ireland was carried out in July 2007. Dinophysis acuta was present as a sub surface layer containing up to 55,000 cells L⁻¹. The population had a high percentage of viable cells (mean: 89%; median: 94%; n = 24) with a high specific division rate (∼0.55 d⁻¹). The layer, of approximately 5 m thickness, did not coincide with the fluorescence maximum and was present as a patch of horizontal dimension less than 10 km × 7 km. Both conventional and towed undulating CTD used in conjunction with high vertical resolution sampling methods showed the patch of Dinophysis to move with a similar speed and direction as the coastal flow, which ran parallel to the coast in the form of a coastal jet with speed of the order of 6.5–7 km day⁻¹. The implications of the alongshore transport of populations of harmful species in coastal jets for monitoring programmes and predictive models are discussed.     

The emergence of Dinophysis acuminata blooms and DSP toxins in shellfish in New York waters

The dynamics of Dinophysis acuminata and its associated diarrhetic shellfish poisoning (DSP) toxins, okadaic acid (OA) and dinophysistoxin-1 (DTX1) as well as pectenotoxins (PTXs), were investigated within plankton and shellfish in Northport Bay, NY, USA, over a four year period (2008–2011). Over the course of the study, Dinophysis bloom densities ranged from ~10⁴ to 10⁶ cells L⁻¹ and exceeded 10⁶ L⁻¹ in 2011 when levels of total OA, total DTX1, and PTX in the water column were 188, 86, and 2900 pg mL⁻¹, respectively, with the majority of the DSP toxins present as esters. These cell densities exceed – by two orders of magnitude – those previously reported within thousands of samples collected from NY waters from 1971 to 1986. The bloom species was positively identified as D. acuminata via scanning electron microscopy and genetic sequencing (cox1 gene). The cox1 gene sequence from the D. acuminata populations in Northport Bay was 100% identical to D. acuminata from Narragansett Bay, RI, USA and formed a strongly supported phylogenetic cluster (posterior probability = 1) that included D. acuminata and Dinophysis ovum from systems along the North Atlantic Ocean. Shellfish collected from Northport Bay during the 2011 bloom had DSP toxin levels (1245 ng g⁻¹ total OA congeners) far exceeding the USFDA action level (160 ng g⁻¹ total OA of shellfish tissue) representing the first such occurrence on the East Coast of the U.S. D. acuminata blooms co-occurred with paralytic shellfish poisoning (PSP) causing blooms of Alexandrium fundyense during late spring each year of the study. D. acuminata cell abundances were significantly correlated with levels of total phytoplankton biomass and Mesodinium spp., suggesting food web interactions may influence the dynamics of these blooms. Given that little is known regarding the combined effects of DSP and PSP toxins on human health and the concurrent accumulation and depuration of these toxins in shellfish, these blooms represent a novel managerial challenge.     

Mass entrapment and lysis of Mesodinium rubrum cells in mucus threads observed in cultures with Dinophysis

The entrapment and death of the ciliate Mesodinium rubrum in the mucus threads in cultures with Dinophysis is described and quantified. Feeding experiments with different concentrations and predator–prey ratios of Dinophysis acuta, Dinophysis acuminata and M. rubrum to study the motility loss and aggregate formation of the ciliates and the feeding behaviour of Dinophysis were carried out. In cultures of either Dinophysis species, the ciliates became entrapped in the mucus, which led to the formation of immobile aggregates of M. rubrum and subsequent cell lysis. The proportion of entrapped ciliates was influenced by the concentration of Dinophysis and the ratio of predator and prey in the cultures. At high cell concentrations of prey (136 cells mL⁻¹) and predator (100 cells mL⁻¹), a maximum of 17% of M. rubrum cells became immobile and went through cell lysis. Ciliates were observed trapped in the mucus even when a single D. acuminata cell was present in a 3.4 mL growth medium. Both Dinophysis species were able to detect immobile or partly immobile ciliates at a distance and circled around the prey prior to the capture with a stretched out peduncle. Relatively high entrapment and lysis of M. rubrum cells in the mucus threads indicates that under certain conditions Dinophysis might have a considerable impact on the population of M. rubrum.     

Harmful effects of Dinophysis to the ciliate Mesodinium rubrum: Implications for prey capture

Toxigenic Dinophysis spp. are obligate mixotrophic dinoflagellates that require a constant supply of prey—Mesodinium rubrum—to achieve long-term growth by means of kleptoplasty. Mesodinium rubrum is, however, a fast moving, jumping ciliate exhibiting an effective escape response from suspensivorous predators. In the present study, a series of laboratory experiments evaluating the motility and survival of M. rubrum in the presence of Dinophysis cells and/or substances contained in their culture medium was designed, in order to assess the mechanisms involved in prey capture by Dinophysis spp. Cell abundance of M. rubrum decreased in the presence of Dinophysis cf. ovum cells producing okadaic acid (OA; up to 7.94 ± 2.67 pg cell⁻¹) and smaller amounts of dinophysistoxin-1 (DTX-1) and pectenotoxin-2 (PTX-2). Prey capture was often observed after the ciliate had been attached to adhesive “mucus traps”, which only appeared in the presence of Dinophysis cells. Before being attached to the mucus traps, M. rubrum cells reduced significantly their swimming frequency (from ∼41 to 19 ± 3 jumps min⁻¹) after only 4 h of initial contact with D. cf. ovum cells. M. rubrum survival was not affected in contact with purified OA, DTX-1 and PTX-2 solutions, but decreased significantly when the ciliate was exposed to cell-free or filtered culture medium from both D. cf. ovum and D. caudata, the latter containing moderate concentrations of free eicosapentaenoic acid and docosahexaenoic acid. The results thus indicate that Dinophysis combines the release of toxic compounds other than shellfish toxins, possibly free PUFAs, and a “mucus trap” to enhance its prey capture success by immobilizing and subsequently arresting M. rubrum cells.     

Environmental control of harmful dinoflagellates and diatoms in a fjordic system

Fjordic coastlines provide an ideal protected environment for both finfish and shellfish aquaculture operations. This study reports the results of a cruise to the Scottish Clyde Sea, and associated fjordic sea lochs, that coincided with blooms of the diarrhetic shellfish toxin producing dinoflagellate Dinophysis acuta and the diatom genus Chaetoceros, that can generate finfish mortalities. Unusually, D. acuta reached one order of magnitude higher cell abundance in the water column (2840 cells L⁻¹) than the more common Dinophysis acuminata (200 cells L⁻¹) and was linked with elevated shellfish toxicity (maximum 601 ± 237 μg OA eq/kg shellfish flesh) which caused shellfish harvesting closures in the region. Significant correlations between D. acuta abundance and that of Mesodinium rubrum were also observed across the cruise transect potentially supporting bloom formation of the mixotrophic D. acuta. Significant spatial variability in phytoplankton that was related to physical characteristics of the water column was observed, with a temperature-driven frontal region at the mouth of Loch Fyne being important in the development of the D. acuta, but not the Chaetoceros bloom. The front also provided important protection to the aquaculture located within the loch, with neither of the blooms encroaching within it. Analysis based on a particle-tracking model confirms the importance of the front to cell transport and shows significant inter-annual differences in advection within the region, that are important to the harmful algal bloom risk therein.     

Growth and loss of distal tissue in blades of Lessonia nigrescens and Lessonia trabeculata (Laminariales)

Meristematic growth and loss of distal tissue from blades of two ecologically important species in the south-east Pacific, Lessonia nigrescens and Lessonia trabeculata, was evaluated during 1 year. Comparative growth was determined by a hole-punch method, loss of distal tissue from the blades was determined by subtracting final blade length (with loss) from expected blade lengths (without loss); growth and tissue loss were transformed to fresh biomass units for calculation of inter-algae differences. The results showed that blade elongation rate increased at the beginning of spring, and declined towards the end of summer, with mean values between 0.40 and 0.08 cm day⁻¹ for L. nigrescens, and 0.65–0.17 cm day⁻¹ for L. trabeculata. Loss of distal tissue varied seasonally when examined as length units for both species; with mean values between 0.24 and 0.10 cm day⁻¹ for L. nigrescens, and 0.51–0.25 cm day⁻¹ for L. trabeculata. Variations in fresh biomass units were only observed in Lessonia trabeculata, increasing in spring, with mean values to 0.13 g (fresh weight) day⁻¹. Annual growth and loss of distal tissue were higher in L. trabeculata (0.41 and 0.39 cm day⁻¹, respectively) than in L nigrescens (0.19 and 0.15 cm day⁻¹). When growth and tissue loss were considered as fresh biomass, monthly gains significantly outweighed loss of distal tissue in both species, but parallel results based on length data followed a different trend. L. trabeculata released about 50% of its growth biomass as particulate organic matter, while the comparative value for L. nigrescens was about 20%.     

Toward predicting Dinophysis blooms off NW Iberia: A decade of events

Dinophysis acuminata and Dinophysis acuta are recurrent species off NW Iberia but their outbreaks occur under different conditions. A decade (2004–2013) of weekly data for each species at two sentinel stations located at the entrance of Rias de Aveiro-AV (NW Portugal, 40°38.6′ N) and Pontevedra-PO (Galicia, Spain, 42°21.5′ N), were used to investigate the regional synchronism and mesoscale differences related to species detection, bloom (>200 cells L⁻¹) initiation and development. Results highlight the high interannual variability of bloom events and summarize the associated meteorological/oceanographic conditions. D. acuta blooms were observed in 2004–2008 and 2013, and the species highest maxima at AV occurred after the highest maxima of its prey Mesodinium, with a time-lag of 2–3 weeks. D. acuminata blooms were observed every year at both stations. The cell concentration time series shows that the blooms generally present a sequence starting in March with D. acuminata in PO and three weeks later in AV, followed by D. acuta that starts at AV and three months later in PO. Exceptionally, D. acuminata blooms occurred earlier at AV than PO, namely in high spring upwelling (2007) or river runoff (2010) years. A four-year gap (2009–2012) of D. acuta blooms occurred after an anomalous 2008 autumn with intense upwelling which is interpreted as the result of an equatorward displacement of the population core. Numerical model solutions are used to analyze monthly alongshore current anomalies and test transport hypotheses for selected events. The results show a strong interannual variability in the poleward/equatorward currents associated with changes in upwelling forcing winds, the advection of D. acuta blooms from AV to PO and the possibility that D. acuminata blooms at AV might result from inocula advected southward from PO. However, the sensitivity of the results to vertical position of the lagrangian tracers call for more studies on species distribution at the various bloom stages.     

Co-occurrence of Dinophysis tripos and pectenotoxins in Argentinean shelf waters

The species Dinophysis tripos is a widely distributed marine dinoflagellate associated with diarrheic shellfish poisoning (DSP) events, which has been recently identified as a pectenotoxin (PTX) producer. In two sampling expeditions carried out during austral autumns 2012 and 2013 along the Argentine Sea (≈38–56° S), lipophilic phycotoxins were measured by tandem mass spectrometry coupled to liquid chromatography (LC–MS/MS) in size-fractionated plankton samples together with microscopic analyses of potentially toxic phytoplankton. PTX-2, PTX-11 and PTX-2sa were recurrently detected in the 50–200 μm fractions, in association to D. tripos. PTX-2 was also widely distributed among the 20–50 μm fractions, mostly related to Dinophysis acuminata. Okadaic acid or its analogs were not detected in any sample. This is the first report of D. tripos related to PTX in the Argentine Sea and the first record of PTX-11 and PTX-2sa for this area. The morphological variability of D. tripos, including the presence of intermediate, small and dimorphic cells, is described. Also, the micro- and mesoplanktonic potential grazers of Dinophysis spp. were explored.     

Escape protein supplementation of growing steers grazing stargrass

A grazing trial utilizing 35 individually supplemented growing steers (211±42 kg initial body weight (BW)) was conducted to study the effect of supplemental escape protein on the performance of steers grazing on stargrass (Cynodon plectostachyus) during the dry season. N in supplements was 100%, 50%, or 0% natural protein (bloodmeal, coconut meal, and soybean meal), and 0%, 50% or 100% urea. All steers received 2 kg of supplement dry matter (DM) (2.2% N) daily during the 90 days of the experiment. Steers fed the urea supplement had the lowest ADG (0.97 kg day⁻¹). There was a linear (P<0.05) response in ADG to the natural protein level (50 and 100%) in supplements containing bloodmeal (1.11 and 1.21 kg day⁻¹) and coconut meal (1.05 and 1.21 kg day⁻¹), but no response was observed with soybean meal (1.01 and 1.0 kg day⁻¹). Forage intake was not affected by supplementation. As a result of the growth response observed for supplements containing bloodmeal and coconut meal above the urea-based and soybean meal supplements, it was concluded that growing ruminants grazing stargrass in the dry season were deficient in escape protein. ©1997 Elsevier Science B.V.     

Climate variability and Dinophysis acuta blooms in an upwelling system

Dinophysis acuta is a frequent seasonal lipophilic toxin producer in European Atlantic coastal waters associated with thermal stratification. In the Galician Rías, populations of D. acuta with their epicentre located off Aveiro (northern Portugal), typically co-occur with and follow those of Dinophysis acuminata during the upwelling transition (early autumn) as a result of longshore transport. During hotter than average summers, D. acuta blooms also occur in August in the Rías, when they replace D. acuminata. Here we examined a 30-year (1985–2014) time series of D. acuta from samples collected by the same method in the Galician Rías. Our main objective was to identify patterns of distribution and their relation with climate variability, and to explain the exceptional summer blooms of D. acuta in 1989–1990. A dome-shaped relationship was found between summer upwelling intensity and D. acuta blooms; cell maxima were associated with conditions where the balance between upwelling intensity and heating, leading to deepened thermoclines, combined with tidal phase (3 days after neap tides) created windows of opportunity for this species. The application of a generalized additive model based on biological (D. acuta inoculum) and environmental predictors (Cumulative June–August upwelling CUI_JJA, average June–August SST_JJA and tidal range) explained more than 70% of the deviance for the exceptional summer blooms of D. acuta, through a combination of moderate (35,000–50,000 m³ s⁻¹ km⁻¹) summer upwelling (CUI_JJA), thermal stratification (SST_JJA > 17 °C) and moderate tidal range (∼2.5 m), provided D. acuta cells (inoculum) were present in July. There was no evidence of increasing trends in D. acuta bloom frequency/intensity nor a clear relationship with NAO or other long-term climatic cycles. Instead, the exceptional summer blooms of 1989–1990 appeared linked to extreme hydroclimatic anomalies (high positive anomalies in SST and NAO index), which affected most of the European Atlantic coast.     

Seasonal variability of lipophilic toxins during a Dinophysis acuta bloom in Western Iberia: Differences between picked cells and plankton concentrates

This work describes and compares the seasonal variability of toxin profiles and content, estimated by LC–MS analyses, in picked cell of Dinophysis acuta Ehrenberg, in plankton concentrates rich in this species, and in extracellular lipophilic toxins collected by adsorbent resins during weekly sampling in a Galician ría (Western Iberia) from October 2005 to January 2006. Picked cells of D. acuta—which exhibited a fairly stable OA:DTX2 ratio, close to 3:2, but a variable okadaates:PTX2 ratio—showed a 9-fold variation in cell toxin quota, which was partly related to cellular volume, with maximum values (19 pg cell⁻¹) observed during the exponential decline of the population. Large differences in toxin profiles and content were observed between picked cells and plankton concentrates (up to 73 pg cell⁻¹ in the latter), that were most conspicuous after the bloom decline. The toxin profile of picked cells was more similar to that observed in the adsorbent resins than to the profiles of plankton concentrates. Their continued detection several weeks after the disappearance of Dinophysis spp. indicates that these toxins may take a long time to be degraded. It is concluded that analyses of picked-cells are essential to determine the contribution of each species of Dinophysis to a toxic outbreak. Estimates of cellular toxin content from plankton concentrates can lead to considerable overestimates after Dinophysis blooms decay due to extracellular toxins that persist in the water column, possibly bound to organic aggregates and detritus, and are retained (>0.22 μm) in the filters.     

Growth of the fungus Paecilomyces lilacinus with n-hexadecane in submerged and solid-state cultures and recovery of hydrophobin proteins

The filamentous fungus Paecilomyces lilacinus was grown on n-hexadecane in submerged (SmC) and solid-state (SSC) cultures. The maximum CO₂ production rate in SmC (V_max = 11.7 mg CO₂ L_g⁻¹ day⁻¹) was three times lower than in SSC (V_max = 40.4 mg CO₂ L_g⁻¹ day⁻¹). The P. lilacinus hydrophobin (PLHYD) yield from the SSC was 1.3 mg PLHYD g protein⁻¹, but in SmC, this protein was not detected. The PLHYD showed a critical micelle concentration of 0.45 mg mL⁻¹. In addition, the PLHYD modified the hydrophobicity of Teflon from 130.1 ± 2° to 47 ± 2°, forming porous structures with some filaments <1 μm and globular aggregates <0.25 μm diameter. The interfacial studies of this PLHYD could be the basis for the use of the protein to modify surfaces and to stabilize compounds in emulsions.     

Production of dinophysistoxin-1 and pectenotoxin-2 by a culture of Dinophysis acuminata (Dinophyceae)

The production of diarrhetic shellfish poisoning toxins (okadaic acid analogues and other lipophilic toxins) by a culture of Dinophysis acuminata, fed with the autotrophic ciliate Myrionecta rubra, was confirmed by LC–MS analysis, and the toxin profile compared with that in the field assemblage of the same species. The growth response of D. acuminata to the density of the food organism was also examined in laboratory experiments. In semi-continuous culture experiments, the growth rates of D. acuminata increased with increasing density of M. rubra and a maximum growth rate of 0.67 per day was calculated. In batch culture experiments; the cellular content of PTX2 and DTX1 were 14.7–14.8 and 2.5–4.8 pg cell^?1, respectively. Okadaic acid, dinophysistoxin-3, pectenotoxin-1, pectenotoxin-6, yessotoxin (YTX) and 45-OHYTX were not detected. PTX2 was detected (cellular toxin content: 22 pg cell^?1), but DTX1 was not detected, in an extract of D. acuminata collected from natural seawater at the same location where the cultured D. acuminata specimens were isolated. These results strongly suggest that D. acuminata produces these toxins during cell growth and that environmental factors influence variations in the toxin composition and specific cellular toxicity.     

Effect of nutrient pulses on photosynthesis of Chaetomorpha linum from a shallow Mediterranean coastal lagoon

The influence of nitrogen and phosphorus pulses on Chaetomorpha linum (Muller) Kutzing growth and photosynthesis was studied in laboratory experiments. Photosynthesis and growth of C. linum from Tancada lagoon seems limited by both nitrogen and phosphorus, as indicated by the high rate (4.7–11.6 mg O₂ g⁻¹ dry weight h⁻¹) of light-saturated photosynthesis (P_m) and growth rates observed under nitrogen plus phosphorus enrichment in relation to enrichment by nitrogen alone (2.9–7.6 mg O₂ g⁻¹ dry weight h⁻¹). Significant increase in nitrogen and phosphorus content as percentage of dry weight was observed in C. linum fertilized with a single nutrient or with nitrogen plus phosphorus. In Tancada lagoon, when availability of nitrogen to primary producers is by pulses, an increase of nitrate concentration in the water column (from 6 to 100 μM) has a greater effect on growth of C. linum (growth rate: 0.13 day⁻¹) than an increase in ammonium concentration (from 20 to 100 μM and growth rate: 0.11 day⁻¹). For a given thallus nitrogen content (0.6–1.4% N), both P_m and the photosynthetic efficiency (α) normalized to dry weight were correlated (r² = 0.73, p < 0.005) indicating that variations in electron transport were coupled to variations in C-fixation capacity. Optimizing both α and P_m may be a general characteristic of thin-structured opportunistic algae in more variable estuarine environments.     

Leaf production and shoot dynamics of Thalassia testudinum by a direct census method

The annual leaf growth and shoot dynamics of Thalassia testudinum were examined in a meadow located near Havana City, Cuba, using direct censuses between January 1995 and January 1996. The net rate of shoot population change, specific shoot recruitment and mortality rates were calculated as the difference between the densities of shoots (tagged or untagged) in consecutive sampling events. The leaf biomass, the daily production, the turnover rate and the rate of leaf biomass loss were also estimated. The estimated mean dry leaf biomass (124.9 ± 9.5 g m⁻²), daily dry leaf production (3.3 ± 0.2 g m⁻² day⁻¹) and turnover rate (2.7 ± 0.1% day⁻¹) were comparable to values previously reported for this species in Cuba and elsewhere. The production of leaves and shoots were higher in spring, declined towards mid summer, and showed the minimum values in January. Shoot recruitment prevailed over shoot mortality from January to March and from July to August, whereas most of the annual shoot mortality occurred between May and July and between August and October. The meadow examined was in close demographic balance along the study period. The results demonstrate that direct census provides reliable estimates of rapid shoot dynamics in T. testudinum.     

Evaluation of the effects of light intensity on growth of the benthic dinoflagellate Ostreopsis sp. 1 using a newly developed photoirradiation-culture system and a novel regression analytical method

Benthic dinoflagellates of the genus Ostreopsis are found all over the world in temperate, subtropical, and tropical coastal regions. Our recent studies revealed that a putative “cryptic” species of Ostreopsis ovata is present widely along Japanese coasts. This organism, Ostreopsis sp. 1, possesses palytoxin analogs and thus its toxic blooms may be responsible for potential toxification of marine organisms. To evaluate the bloom dynamics of Ostreopsis sp. 1, the present study examined the growth responses of Ostreopsis sp. 1 strain s0716 to various light intensities (photon flux densities: μmol photons m⁻² s⁻¹) using a newly devised photoirradiation-culture system. This novel system has white light-emitting diodes (LEDs) capable of more closely simulating the wavelength spectrum of light entering the oceanic water column than do fluorescent tubes and halogen lamps. In this system, the light intensity of the white LEDs was reduced through two polarizing filters by varying the rotation angles of the filters. Thereby, the new system was capable of culturing microalgae under well-controlled light intensity conditions. Ostreopsis sp. 1 grew proportionally when light intensity was increased from 49.5 to 199 μmol photons m⁻² s⁻¹, but its growth appeared to be inhibited slightly at ≥263 μmol photons m⁻² s⁻¹. The relationship between observed growth rates and light intensity was calculated at R > 0.99 (P < 0.01) using a regression analysis with a modified equation of the photosynthesis-light intensity (P-L) model. The equation determined the critical light intensities for growth of Ostreopsis sp. 1 and the organism's growth potential as follows: (1) the threshold light intensity for growth: 29.8 μmol photons m⁻² s⁻¹; (2) the optimum light intensity (L_m) giving the maximum growth rate (μ_max = 0.659 divisions day⁻¹): 196 μmol photons m⁻² s⁻¹; (3) the optimum light intensity range (L_opt) giving ≥95% μ_max: 130–330 μmol photons m⁻² s⁻¹; (4) the semi-optimum range (L_sopt) giving ≥80% μ_max: 90 to over 460 μmol photons m⁻² s⁻¹. The L_sopt represents 4.5–23% ambient light intensity present in surface waters off of a temperate region of the Japanese coast, Tosa Bay; putatively, this semi-optimum range of light intensity appears at depth of 12.9–27.8 m. Considering these issues, our data indicate that Ostreopsis sp. 1 in coastal environments may form blooms at ca. ∼28 m depth in regions along Japanese coasts.     

Germination fluctuation of toxic Alexandrium fundyense and A. pacificum cysts and the relationship with bloom occurrences in Kesennuma Bay,Japan

While cyst germination may be an important factor for the initiation of harmful/toxic blooms, assessments of the fluctuation in phytoplankton cyst germination, from bottom sediments to water columns, are rare in situ due to lack of technology that can detect germinated cells in natural bottom sediments. This study introduces a simple mesocosm method, modeled after previous in situ methods, to measure the germination of plankton resting stage cells. Using this method, seasonal changes in germination fluxes of toxic dinoflagellates resting cysts, specifically Alexandrium fundyense (A. tamarense species complex Group I) and A. pacificum (A. tamarense species complex Group IV), were investigated at a fixed station in Kesennuma Bay, northeast Japan, from April 2014 to April 2015. This investigation was conducted in addition to the typical samplings of seawater and bottom sediments to detect the dinoflagellates vegetative cells and resting cysts. Bloom occurrences of A. fundyense were observed June 2014 and February 2015 with maximum cell densities reaching 3.6 × 10⁶ cells m⁻² and 1.4 × 10⁷ cells m⁻², respectively. The maximum germination fluxes of A. fundyense cysts occurred in April 2014 and December 2014 and were 9.3 × 10³ cells m⁻² day⁻¹ and 1.4 × 10⁴ cells m⁻² day⁻¹, respectively. For A. pacificum, the highest cell density was 7.3 × 10⁷ cells m⁻² during the month of August, and the maximum germination fluxes occurred in July and August, reaching 5.8 × 10² cells m⁻² day⁻¹. Thus, this study revealed the seasonal dynamics of A. fundyense and A. pacificum cyst germination and their bloom occurrences in the water column. Blooms occurred one to two months after peak germination, which strongly suggests that both the formation of the initial population by cyst germination and its continuous growth in the water column most likely contributed to toxic bloom occurrences of A. fundyense and A. pacificum in the bay.     

Marine snow formation by the toxin-producing diatom,Pseudo-nitzschia australis

The formation of marine snow (MS) by the toxic diatom Pseudo-nitschia australis was simulated using a roller table experiment. Concentrations of particulate and dissolved domoic acid (pDA and dDA) differed significantly among exponential phase and MS formation under simulated near surface conditions (16 °C/12:12-dark:light cycle) and also differed compared to subsequent particle decomposition at 4 °C in the dark, mimicking conditions in deeper waters. Particulate DA was first detected at the onset of exponential growth, reached maximum levels associated with MS aggregates (1.21 ± 0.24 ng mL⁻¹) and declined at an average loss rate of ∼1.2% pDA day⁻¹ during particle decomposition. Dissolved DA concentrations increased throughout the experiment and reached a maximum of ∼20 ng mL⁻¹ at final sampling on day 88. The succession by P. australis from active growth to aggregation resulted in increasing MS toxicity and based on DA loading of particles and known in situ sinking speeds, a significant amount of toxin could have easily reached the deeper ocean or seafloor. MS formation was further associated with significant dDA accumulation at a ratio of pDA: dDA: cumulative dDA of approximately 1:10:100. Overall, this study confirms that MS functions as a major vector for toxin flux to depth, that Pseudo-nitzschia-derived aggregates should be considered ‘toxic snow’ for MS-associated organisms, and that effects of MS toxicity on interactions with aggregate-associated microbes and zooplankton consumers warrant further consideration.     

Toxigenic phytoplankton and concomitant toxicity in the mussel Choromytilus meridionalis off the west coast of South Africa

The variability of toxigenic phytoplankton and the consequent uptake and loss of toxins by the mussel Choromytilus meridionalis was investigated in the southern Benguela at the event scale (3–10 days) in response to the upwelling–downwelling cycle. Phytoplankton and mussel samples were collected daily (20 March–11 April 2007) from a mooring station (32.04°S; 18.26°E) located 3.5 km offshore of Lambert's Bay, within the St Helena Bay region. Rapid changes in phytoplankton assemblages incorporated three groups of toxigenic phytoplankton: (1) the dinoflagellate Alexandrium catenella; (2) several species of Dinophysis, including Dinophysis acuminata, Dinophysis fortii, Dinophysis hastata and Dinophysis rotundata; and (3) members of the diatom genus Pseudo-nitzschia. Analysis of phytoplankton concentrates by LC–MS/MS or LC-FD provided information on the toxin composition and calculated toxicity of each group. Several additional in vitro assays were used for the analysis of toxins in mussels (ELISA, RBA, MBA for PSP toxins; and ELISA for DSP toxins). Good correspondence was observed between methods except for the MBA, which provided significantly lower (approximately 2-fold) estimates of PSP toxins. PSP and DSP toxins both exceeded the regulatory limits in Choromytilis meridionalis, but ASP toxins were undetected. Differences were observed in the composition of both PSP and DSP toxins in C. meridionalis from that of the ingested dinoflagellates (PSP toxins showed an increase in STX, C1,2, and traces of dcSTX and GTX1,4 and a decrease in NEO; DSP toxins showed an increased in DTX1, and traces of PTX2sa, and a decrease in OA). The rate of loss of PSP toxins following dispersal of the A. catenella boom was 0.12 d⁻¹. Variation in the loss rates of different PSP toxins contributed to the change in toxin profile in C. meridionalis. Prediction of net toxicity in shellfish of the nearshore environment in the southern Benguela is limited due to rapid phytoplankton community changes, high variability in cellular toxicity, and the selective uptake and loss of toxins, and/or transformation of toxins.