Pg Domoic Acid Research Articles

Spatial distribution and source of biotoxins in phytoplankton from the South China Sea, China

Marine phycotoxins severely threaten ecosystem health and mariculture. This study investigates the spatial distribution and source of diverse phycotoxins in the South China Sea (SCS), during four 2019/2020 cruises. Saxitoxin (STX) and okadaic acid (OA) -groups, azaspiracids, cyclic imines, pectenotoxins (PTX), yessotoxins, and domoic acid (DA) toxins were analyzed in microalgal samples. PTX2 occurred with the highest (93.5%) detection rate (DR) during all cruises, especially in the Pearl River Estuary (PRE) in June 2019. Homo-yessotoxin (hYTX) and DA were found during three cruises in August 2020, and high DR of hYTX (67.7%, 29.3%) and DA (29.0%, 29.3%) in the PRE and Guangdong coast, respectively, in June 2019 and 2020, peaking at concentrations of 777 pg hYTX L-1 and 38514 pg DA L-1. The phycotoxin distribution demonstrated that DA-producing microalgae gathered close to the PRE and Guangdong coast, while hYTX-producing microalgae distributed relatively far offshore. Microalgae producing PTX2- and STX-group toxins were more widely living in the SCS. High-throughput sequencing results suggested that Alexandrium pacificum and Gonyaulax spinifera were responsible for STX-group toxins and hYTX, respectively, while Pseudo-nitzschia cuspidata was the main source of DA. Widely distributed PTX2, hYTX, and DA were reported for the first time in the SCS.

Organic polymer consumption facilitates domoic acid entry into the marine food web without direct ingestion of Pseudo-nitzschia

Domoic acid (DA) is a neurotoxin produced by diatoms from the genera Pseudo-nitzschia and Nitzschia. DA is transferred through the food web when consumed by organisms such as copepods (e.g., Acartia tonsa). DA bioaccumulates in higher trophic levels and poses a threat to human health through amnesic shellfish poisoning. Laboratory experiments using a DA reference standard demonstrated that mild turbulence facilitates formation of organic polymer aggregates >0.6 µm in-vivo that can scavenge dissolved DA (dDA). Using A. tonsa, we demonstrate that DA can be assimilated through consumption of these organic polymers which scavenged dDA —a pathway which does not require direct ingestion of the toxin-producer Pseudo-nitzschia. In filtered seawater with spiked DA, copepods accumulated 24.8 ± 4.7 pg DA copepod−1 (2.1 ppm) on average by consuming organic polymers. This was validated in one out of five experiments using ambient DA concentrations. Copepods were suspended in particle-free seawater and accumulated 14.4 ± 3.8 pg DA copepod−1 (1.20 ppm), and in particle-concentrated seawater they accumulated 40.9 ± 3.8 pg DA copepod−1 (3.42 ppm). Data from this experiment suggests that ~34% of the total assimilated DA entered via an organic polymer-bound DA pathway. This experiment had the highest Pseudo-nitzschia spp. abundance (~225,000 cells L − 1) and cellular toxin quota, up to 0.88 pg DA cell−1, relative to the other four ambient DA experiments. These results demonstrate the potential for DA to enter the marine food web through an alternate pathway and may have considerable implications to understanding the flow of DA through marine food webs, and how we monitor DA and its potential vectors into the food web.

Fifteen years of Pseudo-nitzschia in an Australian estuary, including the first potentially toxic P. delicatissima bloom in the southern hemisphere

In March 2018, an unprecedented bloom of a species of Pseudo-nitzschia occurred in Berowra Creek, a major tributary of the Hawkesbury River estuary, New South Wales, Australia (max cell abundance 5.7 × 106 cells L−1). Morphological and phylogenetic analysis showed the bloom was dominated by the species P. delicatissima, with toxin characterization by LC-MS confirming one of the seven strains isolated produced domoic acid (DA), at a concentration of 0.34 pg DA per cell.In response to this unprecedented bloom, we used a fifteen year series of phytoplankton and physico-chemical data to relate environmental factors to spatial and temporal variability of Pseudo-nitzschia species throughout the estuary. Species in the Pseudo-nitzschia delicatissima group consistently dominated all locations monitored throughout the estuary, with peak cell densities observed in the austral autumn (lowest in winter). Overall, Generalised Additive Models (GAMs) suggested that high cell concentrations of Pseudo-nitzschia were linked to an increase in soluble reactive phosphorus and a decrease in total nitrogen at all sites, with up to 55% of the deviance explained. Twenty species of Pseudo-nitzschia have now been reported in Australian coastal waters, four of which are confirmed DA producers. We hypothesise that increased urbanization, nutrient input and warmer waters could combine to increase Pseudo-nitzschia blooms in south-eastern Australia.

Evaluation of intracellular and extracellular domoic acid content in Pseudo-nitzschia multiseries cell cultures under different light regimes

Pseudo-nitzschia multiseries is a diatom species associated with the production of domoic acid (DA), a water soluble neurotoxin that is easily transferred up in the food web, causing devastating effects on top marine organisms and humans. Despite studies on Pseudo-nitzschia are relevant to human health safety, partitioning of marine toxins between intracellular and extracellular fractions are poorly documented. This study aimed to determine the growth rates and DA content, both intracellular and extracellular, of Pseudo-nitzschia multiseries cultures at three different light settings (15, 120 and 560 μmol m−2 s−1). The optimal conditions for cell growth were observed at 120 and 560 μmol m−2 s−1, whereas DA production was observed in P. multiseries at 15 and 120 μmol m−2 s−1, ranging between 0.18-2.56 and 0.16–3.5 pg DA cell−1, respectively. Higher intracellular DA concentrations were found during the senescence phase at low light intensity and during the exponential phase at medium light intensity, while higher concentrations of dissolved DA were found at low and medium light intensities in the senescence phase reaching 3 and 10 ng DA mL−1 respectively. The amount of toxin released into the culture medium represents the most important fraction ranging between 63 and 98% during the exponential phase and nearly 99% during the senescence phase. In contrast, under low light intensity, dissolved DA was detected in the culture medium only during the senescence phase. This study confirms the importance of light intensity on DA production and clearly shows that dissolved domoic acid is an important fraction in Pseudo-nitzschia cultures, suggesting with the careful assumptions of results from static cultures extrapolated to bloom situations that waterborne exposure of marine organism should be considered during blooms of Pseudo-nitzschia multiseries.

Morphological, molecular and toxigenic characteristics of Namibian Pseudo-nitzschia species – including Pseudo-nitzschia bucculenta sp. nov.

A field study was undertaken to investigate the occurrence and toxin production of species in the diatom genus Pseudo-nitzschia in Namibian waters, in the extremely productive Benguela upwelling system. From surveys conducted on the R/V Mirabilis and the R/V !Anichab, 52 strains were morphologically determined to species level, supported by nuclear ITS rDNA data. Seven species were identified; P. australis, P. decipiens, P. dolorosa, P. fraudulenta, P. plurisecta, P. pungens var. cingulata, and the new species P. bucculenta F. Gai, C. K. Hedemand, N. Lundholm & Ø. Moestrup sp. nov.Molecular and morphological diversity of the Namibian Pseudo-nitzschia species is discussed. Most importantly, P. bucculenta is both morphologically and phylogenetically most similar to P. dolorosa differing mainly in valve width and densities of striae, poroids and band striae as well as by four hemi-compensatory base changes in the ITS2. Morphological and molecular differences among the strains of P. decipiens suggest a temperate and a warm water subdivision. The geographical and toxigenic characteristics of the identified Pseudo-nitzschia species are described and compared to previous studies. Initial tests of toxin production in all seven species revealed production of domoic acid (DA) in two species: one strain of P. australis (0.074 pg DA cell−1) and two strains of P. plurisecta (0.338 pg DA cell−1 and 0.385 pg DA cell−1).

Detection of domoic acid in Mytilus galloprovincialis and Ostrea edulis linked to the presence of Nitzschia bizertensis in Bizerte Lagoon (SW Mediterranean)

Seasonal variation of DA in bivalves was monitored from August 2008 to March 2009 at a shellfish station in Bizerte Lagoon, one of the most important shellfish areas in Tunisia. At the same time, dynamics of potentially toxic diatoms was studied at six stations in the lagoon. Several diatom strains were isolated, maintained in culture and their identification as well as the toxicity was examined.During the study period, domoic acid (DA) was detected in Tunisia, in both mussels (Mytilus galloprovincialis: 0.13–0.86 μg DA g−1 tissue) and oysters (Ostrea edulis: 0.42–1.04 μg DA g−1 tissue). The amount of DA in the two species was below the EU guideline limits for shellfish (20 μg DA g−1).Within the phytoplankton community, potentially toxic diatoms were observed in high densities (104–106 cells l−1). Among the 9 established cultured strains, seven were identified as the recently described species Nitzschia bizertensis sp. nov, which is apparently a commonly occurring species in the Bizerte Lagoon. Among tested five strains of N. bizertensis, four were confirmed to be toxin producers, containing 2–7 × 10−3 pg DA cell−1 during stationary growth phase.Our results indicate for the first time a Nitzschia species, here Nitzschia bizertensis, could be in relation to accumulation of DA in shellfish. Monitoring of DA in phytoplankton and shellfish should be considered for Bizerte Lagoon in order to gain knowledge about the ecology and toxin-production of N. bizertensis and to explore the risk of DA accumulation in the local shellfish industry caused by this species.

The diatom genus Pseudo-nitzschia (Bacillariophyceae) in New South Wales, Australia: morphotaxonomy, molecular phylogeny, toxicity, and distribution.

Species belonging to the potentially harmful diatom genus Pseudo-nitzschia, isolated from 16 localities (31 sampling events) in the coastal waters of south-eastern Australia, were examined. Clonal isolates were characterized by (i) light and transmission electron microscopy; (ii) phylogenies, based on sequencing of nuclear-encoded ribosomal deoxyribonucleic acid (rDNA) regions and, (iii) domoic acid (DA) production as measured by liquid chromatography-mass spectrometry (LC-MS/MS). Ten taxa were unequivocally confirmed as Pseudo-nitzschia americana, P.arenysensis, P.calliantha, P.cuspidata, P.fraudulenta, P.hasleana, P.micropora, P.multiseries, P.multistriata, and P.pungens. An updated taxonomic key for south-eastern Australian Pseudo-nitzschia is presented. The occurrence of two toxigenic species, P.multistriata (maximum concentration 11pg DA per cell) and P.cuspidata (25.4pg DA per cell), was documented for the first time in Australia. The Australian strains of P.multiseries, a consistent producer of DA in strains throughout the world, were nontoxic. Data from 5,888 water samples, collected from 31 oyster-growing estuaries (2,000km coastline) from 2005 to 2009, revealed 310 regulatory exceedances for "Total Pseudo-nitzschia," resulting in six toxic episodes. Further examination of high-risk estuaries revealed that the "P.seriata group" had highest cell densities in the austral summer, autumn, or spring (species dependent), and lowest cell densities in the austral winter, while the "P.delicatissima group" had highest in winter and spring.

Growth and domoic acid content of Pseudo-nitzschia australis isolated from northwestern Baja California, Mexico, cultured under batch conditions at different temperatures and two Si:NO 3 ratios

Domoic acid (DA) poisoning in the southern part of the California Current System has been associated typically with blooms of Pseudo-nitzschia australis. The environmental variables that promote growth and DA production in the Mexican part of this system have not been identified. The present study investigated the effect of temperature and two nutrient ratios on the growth characteristics and DA content of two (BTS-1, BTS-2) P. australis strains isolated from the Pacific coast of northern Baja California peninsula, México. Of the different temperatures assayed (10, 12, 14, 15, 18 and 20 °C), the maximum cell abundance was detected at 12 °C for BTS-2 and 14 °C for BTS-1. The highest maximum specific growth rate (1.69 day −1) was measured at 15 °C for BTS-2. With the exception of cells maintained at 15 °C, growth characteristics were similar in P. australis cultured in a high Si:NO 3 (2.5) or low Si:NO 3 (0.5) ratio at each temperature. Dissolved (dDA) and cellular (cDA) DA content measured at the stationary phase of growth was similar in cells cultivated at the different temperatures. No difference in cDA (between 0.11 and 1.87 pg DA cell −1) was observed in cells cultivated at the two nutrient ratios. To evaluate if P. australis accumulates DA (cDA + dDA) at different stages of the culture and not only during the stationary phase of growth, the BTS-1 strain was cultivated at 14 °C and the content of this toxin was measured during culture development. The cultures were maintained at high (HL; 200 μmol quanta m −2 s −1) and low light (LL; 30 μmol quanta m −2 s −1) and in the two nutrient ratios to evaluate the effect of these variables on DA content. The photosynthetic performance and pigment concentration were measured as indicators of the physiological condition of the cells. cDA was detected in all culture conditions and during the different stages of growth. The highest DA content was measured during the lag phase of growth and it was present mainly in the medium (dDA = 70.83 pg DA cell −1). Cells cultivated at HL produced more DA than LL cultured cells. P. australis cultured in HL presented lower photosynthetic rates than LL cells and had similar concentrations of photoprotective pigments and the highest maximum photosynthetic rates were detected during the lag phase of growth in all culture conditions. The results demonstrate that P. australis from northern Baja California peninsula presents a narrow temperature range for optimal growth under batch culture conditions. P. australis produce DA at different stages of growth, and DA content was related to the light intensity at which the cells were cultivated.

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas

Oceanic high-nitrate, low-chlorophyll environments have been highlighted for potential large-scale iron fertilizations to help mitigate global climate change. Controversy surrounds these initiatives, both in the degree of carbon removal and magnitude of ecosystem impacts. Previous open ocean enrichment experiments have shown that iron additions stimulate growth of the toxigenic diatom genus Pseudonitzschia. Most Pseudonitzschia species in coastal waters produce the neurotoxin domoic acid (DA), with their blooms causing detrimental marine ecosystem impacts, but oceanic Pseudonitzschia species are considered nontoxic. Here we demonstrate that the sparse oceanic Pseudonitzschia community at the high-nitrate, low-chlorophyll Ocean Station PAPA (50 degrees N, 145 degrees W) produces approximately 200 pg DA L(-1) in response to iron addition, that DA alters phytoplankton community structure to benefit Pseudonitzschia, and that oceanic cell isolates are toxic. Given the negative effects of DA in coastal food webs, these findings raise serious concern over the net benefit and sustainability of large-scale iron fertilizations.

Crassostrea virginica grazing on toxic and non-toxic diatoms

Despite high abundances of toxic Pseudo-nitzschia spp. over Louisiana oyster beds ( Crassostrea virginica; eastern oyster) there have been no documented cases of amnesic shellfish poisoning (ASP) in the state. Two possible explanations are that oysters do not readily feed on long pointed chains of Pseudo-nitzschia cells or they discriminate against toxic cells while grazing. To test these hypotheses, short-term grazing experiments were conducted with several diatoms, including the domoic acid (DA)-producing Pseudo-nitzschia multiseries (1.31 ± 0.057 pg DA cell −1) and the non-toxic Pseudo-nitzschia delicatissima, Thalassiosira weissflogii, and Ditylum brightwellii. Grazing rates on the small centric species T. weissflogii were significantly higher than on the larger and pointier D. brightwellii and either Pseudo-nitzschia species. Grazing on toxic P. multiseries and non-toxic P. delicatissima was not significantly different. Pseudofeces production was higher and feces production was occasionally lower in oysters fed Pseudo-nitzschia spp. than in oysters fed the other two diatoms. Our data demonstrate lower filtration rates of C. virginica on Pseudo-nitzschia spp. relative to the other diatoms tested and comparable filtration on toxic and non-toxic Pseudo-nitzschia spp. These findings suggest that eastern oysters do not discriminate amongst food types due to DA content.

Accumulation of domoic acid and its effect on juvenile king scallop Pecten maximus (Linnaeus, 1758)

Juvenile king scallops Pecten maximus of 1.23 cm shell height were exposed to domoic acid (DA) and the toxin accumulation and effects of bearing this toxin were investigated. After feeding on a formulated feed containing about 0.19 pg DA per particle for 7 days, the juvenile scallops incorporated a relatively low DA burden of 15.9 ng g − 1 , which was similar to those exposed to 10 and 50 ng ml − 1 dissolved DA. The maximum DA accumulation, 302.5 ng g − 1 whole tissue, was achieved by scallops maintained in 100 ng ml − 1 dissolved DA for 18 days. No significant effect of exposure to DA was observed on feeding rate (on microalgae), shell valve activity (shell clatter), righting response, or the secretion of byssus threads. However, negative impacts on growth rate and survival were found for scallops exposed to DA-spiked feed. This is the first study on DA accumulation in juvenile P. maximus and on the negative effect of DA on this life history stage.

Distribution, Abundance and Domoic Acid Analysis of the Toxic Diatom Genus Pseudo-nitzschia from the Chesapeake Bay

Very little research has been conducted on mid-Atlantic estuarine populations of the diatom Pseudo-nitzschia despite recent evidence of toxicity in regional isolates. We collected field samples from the Chesapeake Bay region from 2002 to 2007 for Pseudo-nitzschia enumeration and toxin analysis. Abundances of Pseudo-nitzschia were highest in the winter and spring at ∼103 cells ml−1. Domoic acid (DA) was detectable in 42% of samples tested, but concentrations were generally low, ranging from 0 to 1,037 pg DA ml−1 (mean 176 pg DA ml−1), and DA cell quota ranged from <0.1 to 49.4 pg DA per cell (mean 1.4 pg DA per cell). Although Pseudo-nitzschia populations were observed year round when salinity was ≥5, populations were highest from February to May when temperatures were low (2–15°C) and salinity relatively high (≥10). Six species of Pseudo-nitzschia were identified via transmission electron microscopy of the samples: P. pungens (Grunow ex Cleve) Hasle, P. calliantha Lundholm, Moestrup et Hasle, P. subpacifica (Hasle) Hasle, P. cuspidata (Hasle) Hasle emend. Lundholm, Moestrup et Hasle, P. fraudulenta (Cleve) Hasle and P. multiseries (Hasle) Hasle. P. calliantha was the most common and not previously reported from the Chesapeake Bay. Of these species, P. pungens, P. calliantha, P. cuspidata, P. fraudulenta and P. multiseries are known to produce domoic acid.

Domoic acid production by Pseudo-nitzschia calliantha Lundholm, Moestrup et Hasle (bacillariophyta) isolated from the Black Sea

A species of Pseudo-nitzschia isolated from Sevastopol Bay, Black Sea, was examined for its toxicity. The species was identified as P. calliantha Lundholm, Moestrup et Hasle based on SEM and TEM examination. Domoic acid (DA) was detected in batch culture throughout the growth cycle of P. calliantha. The production of DA by this diatom species was confirmed by fluorenylmethoxycarbonyl (FMOC) derivatization and HPLC-fluorescence method. The cellular DA level was higher in the early exponential phase, with the maximum value of 0.95 pg DA cell −1. In the stationary phase, the cellular DA levels declined. This is the first record of a DA producing diatom isolated from the Black Sea.

Ingestion of domoic acid and its impact on king scallop (Pecten maximus, Linnaeus 1758)

A simple method for spiking formulated feed with domoic acid (DA) was developed in this study. DA feed was prepared by mixing 0.15 mL 100 μg mL−1 DA with 0.1 g formulated feed, and drying the mixture at room temperature for 2h. The prepared DA feed contained 0.19 pg DA per particle. Of the added DA, 46.72% was retained in the feed. Relatively high DA retention (about 50%) was recorded after DA feed was soaked in water for 2h. Exposure to DA feed for 7 d did not cause the increase of tissue DA level of adult king scallop (Pecten maximus) significantly in 60 d. The increase of their gonad index after DA exposure was not significantly different from the control. No significant change in DA level was found in spermary, ovary or fertilized eggs after DA exposure. These results indicated that DA excretion may be more efficient than DA accumulation under the current experimental conditions, and the mechanism of domoic acid incorporation in P. maximus may involve intracellular biotransformation.

Blooms of Pseudo-nitzschia and domoic acid in the San Pedro Channel and Los Angeles harbor areas of the Southern California Bight, 2003–2004

Abundances of Pseudo-nitzschia spp. and concentrations of particulate domoic acid (DA) were determined in the Southern California Bight (SCB) along the coasts of Los Angeles and Orange Counties during spring and summer of 2003 and 2004. At least 1500 km 2 were affected by a toxic event in May/June of 2003 when some of the highest particulate DA concentrations reported for US coastal waters were measured inside the Los Angeles harbor (12.7 μg DA L −1). Particulate DA levels were an order of magnitude lower in spring of 2004 (February and March), but DA concentrations per cell at several sampling stations during 2004 exceeded previously reported maxima for natural populations of Pseudo-nitzschia (mean = 24 pg DA cell −1, range = 0–117 pg DA cell −1). Pseudo-nitzschia australis dominated the Pseudo-nitzschia assemblage in spring 2004. Overall, DA-poisoning was implicated in >1400 mammal stranding incidents within the SCB during 2003 and 2004. Ancillary physical and chemical data obtained during our regional surveys in 2004 revealed that Pseudo-nitzschia abundances, particulate DA and cellular DA concentrations were inversely correlated with concentrations of silicic acid, nitrogen and phosphate, and to specific nutrient ratios. Particulate DA was detected in sediment traps deployed at 550 and 800 m depth during spring of 2004 (0.29–7.6 μg DA (g sediment dry weight) −1). The highest DA concentration in the traps was measured within 1 week of dramatic decreases in the abundances of Pseudo-nitzschia in surface waters. To our knowledge these are the deepest sediment trap collections from which DA has been detected. Sinking of the spring Pseudo-nitzschia bloom may constitute a potentially important link between DA production in surface waters and benthic communities in the coastal ocean near Los Angeles. Our study indicates that toxic blooms of Pseudo-nitzschia are a recurring phenomenon along one of the most densely populated coastal stretches of the SCB and that the severity and magnitude of these events can be comparable to or greater than these events in other geographical regions affected by domoic acid.

DOMOIC ACID PRODUCTION By PSEUDO‐NITZSCHIA SERIATA (BACILLARIOPHYCEAE) IN SCOTTISH WATERS1

In 1999, a 49,000 km2 area in western Scottish waters was closed to shellfish harvesting due to the amnesic shellfish poisoning (ASP) toxin domoic acid (DA). The only previously confirmed DA producer identified had been Pseudo‐nitzschia australis Frenguelli. The toxin has appeared every year since and has led to more harvesting closures. We isolated and cultured two strains of Pseudo‐nitzschia seriata f. seriata (P. T. Cleve) H. Peragallo from western Scottish waters in 2001 and 2002. They were identified using TEM analysis of their morphological fine structure and sequencing of the internal transcribed spacer (ITS)1, 5.8S, ITS2, and partial large subunit (LSU) rDNA. The morphology of the Scottish P. seriata f. seriata strains differed slightly, for example, in the number of poroid rows, from descriptions in identification keys. Comparison of P. seriata sequences with those of two co‐occurring Pseudo‐nitzschia australis isolates showed an overall divergence of only 0.012. Sequence divergence between both species was highest in the ITS1 region (0.036). Combined morphological and genetic approaches are needed to identify closely related Pseudo‐nitzschia species. The P. seriata strains grew successfully at 15°C, suggesting that although seen as a psychrophilic species, it may also occur at higher water temperatures. All isolates produced DA in stationary phase (measured on day 25): 0.16–0.23 pg DA·cell−1 in P. seriata and 0.15–1.68 pg DA·cell−1 in P. australis. Our study is the first to identify P. seriata f. seriata as a DA producer in Scottish waters and indicates that at least it and P. australis can be responsible for ASP toxicity in that region.

CONFIRMATION OF DOMOIC ACID PRODUCTION BY PSEUDO‐NITZSCHIA AUSTRALIS (BACILLARIOPHYCEAE) ISOLATED FROM IRISH WATERS1

A nonaxenic isolate of the potentially toxic diatom Pseudo‐nitzschia australis (Frenguelli) from Irish waters was tested in two separate batch culture experiments. When grown under a low irradiance (∼12 μmol photons·m−2·s−1; 16:8‐h light:dark cycle) for up to 40 days, the culture produced only trace amounts of the neurotoxin domoic acid (DA) during late stationary phase. Growth at a higher irradiance (∼115 μmol photons·m−2·s−1; 12:12‐h light:dark cycle) resulted in DA production starting during late exponential phase and reaching a maximum concentration of 26 pg DA·cell−1 during late stationary phase. Liquid chromatography coupled to mass spectrometry was used to confirm the identity of DA in the culture. Irradiance and photoperiod could be important factors that contribute directly or indirectly to the control of DA production in P. australis. This is the first record of a DA‐producing diatom in Irish waters, and results indicate P. australis may have been the source of DA that has recently contaminated shellfisheries in this area.

Accumulation of domoic acid by the sea scallop (Placopecten magellanicus) fed cultured cells of toxic Pseudo-nitzschia multiseries

Sea scallops (Placopecten magellanicus) were fed Pseudo-nitzschia multiseries (formerly P. pungens f. multiseries, Nitzschia pungens f. multiseries) cells of high domoic acid (DA) content (4.0-6.7 pg DA cdot cell-1) for 22 days, followed by 14 days of feeding with nontoxic microalgae. DA was incorporated within 24 h by the scallops, with increased uptake after 6 days, and was concentrated in tissues in the following order: digestive gland &gt;&gt; remaining soft tissue &gt;&gt; adductor muscle. A maximum DA concentration of 3108 mu g cdot g-1 was recorded in the digestive gland, approximately 150 times the regulatory limit (20 mu g DA cdot g-1) and among the highest levels observed in bivalve molluscs; however, only trace amounts, 0.7-1.5 mu g cdot g-1, were found concomitantly in the adductor muscle. At the end of the exposure period, 50.9% of the DA that had been supplied to the scallops had been incorporated into the tissues. Concentrations in the digestive gland 14 days after termination of the toxic diet remained high, 752 mu g DA cdot g-1. Throughout the experiment, there was no sign of illness or mortality attributable to high DA loading, although the destructive sampling of animals did not allow us to assess the effects of the toxin in the longer term.

CHANGES IN DOMOIC ACID PRODUCTION AND CELLULAR CHEMICAL COMPOSITION OF THE TOXIGENIC DIATOM PSEUDO‐NITZSCHIA MULTISERIES UNDER PHOSPHATE LIMITATION1

ABSTRACTProduction of domoic acid (DA), a neurotoxin, by the diatom Pseudo‐nitzschia multiseries (previously Nitzschia pungens f. multiseries) Hasle and its cellular chemical composition were studied in phosphate‐limited chemostat continuous cultures and in subsequent batch cultures. Under steady‐state chemostat conditions, DA production increased from 0.01 to 0.26 pg DA · cell−1· d−1 as the growth rate decreased. When the nutrient supply was discontinued (to produce a batch culture), DA production was enhanced by a factor of ca. 3. DA production was temporarily suspended upon addition of phosphate to the batch cultures but resumed 1 d later at a higher rate coincident with the decline of phosphate uptake. In both steady‐state continuous culture and batch culture, more DA was produced when alkaline phosphatase activity (APA) was high. The association of high DA production with high levels of APA and high cellular N:P ratios strongly suggests that phosphate limitation enhances DA production. Also, DA production was high when other primary metabolism (e.g. uptake of carbon, nitrogen, phosphorus and silicon, and cell division) was low, but chlorophyll a and adenosine triphosphate were generally high. This suggests that the synthesis of DA requires a substantial amount of biogenic energy.

Effects of silicate limitation on production of domoic acid, a neurotoxin, by the diatom Pseudo-nitzschia multiseries. II. Continuous culture studies

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 131:235-243 (1996) - doi:10.3354/meps131235 Effects of silicate limitation on production of domoic acid, a neurotoxin, by the diatom Pseudo-nitzschia multiseries. II. Continuous culture studies Pan Y, Subba Rao DV, Mann KH, Li WKW, Harrison WG Production of domoic acid (DA) by Pseudo-nitzschia multiseries (Hasle) was studied using continuous cultures with growth rates ranging from 0.06 to 0.67 d-1. At steady states, DA concentrations were 1.65 to 553.20 ug l-1 and production rates were 0.007 to 1.354 pg DA cell-1 d-1. Both were negatively correlated with rates of growth and silicate uptake. DA production was studied further by stopping the addition of fresh medium, thus producing batch mode experiments, in some of which silicate was allowed to decline, while in another silicate was increased to 85 uM. In those where silicate declined, DA production increased by a factor of 3. The maximum production rate attained was 3.17 pg DA cell-1 d-1 and the highest DA concentration in the culture was 768.5 ug DA l-1, of which 664 ug l-1 was in the cells (11.9 pg DA cell-1). In the experiment where silicate was enriched, DA production was suspended soon after the enrichment, but resumed when silicate in the medium became low. The results suggested differences in kinetics of DA production and growth under different supply rates of silicate. There appear to be 2 types of conditions associated with DA production. When dissolved silicate is moderately low and there is a decline in overall physiological activity, intrinsic factors probably trigger the formation of a moderate amount of DA. When dissolved silicate is severely limiting, the extrinsic stress leads to considerably enhanced production of DA. Pseudo-nitzschia multiseries . Domoic acid . Silicate limitation . Chemostat . Continuous cultures Full text in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 131. Publication date: February 08, 1996 Print ISSN:0171-8630; Online ISSN:1616-1599 Copyright © 1996 Inter-Research.