Limnobacter alexandrii sp. nov., a thiosulfate-oxidizing, heterotrophic and EPS-bearing Burkholderiaceae isolated from cultivable phycosphere microbiota of toxic Alexandrium catenella LZT09.

A novel Gram-negative, aerobic, motile and short rod-shaped bacterium with exopolysaccharides production, designated as LZ-4T, was isolated from cultivable phycosphere microbiota of harmful algal blooms-causing marine dinoflagellate Alexandrium catenella LZT09 which produces paralytic shellfish poisoning toxins. Strain LZ-4T was able to use thiosulfate (optimum concentration 10 mM) as energy source for bacterial growth. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain LZ-4T belonged to the genus Limnobacter, showing high 16S rRNA gene sequences similarities with L. thiooxidans DSM 13612T (99.4%), L. humi NBRC 11650T (98.2%) and L. litoralis NBRC 105857T (97.2%), respectively. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between LZ-4T and L. thiooxidans DSM 13612T were 78.9 and 21.9%, respectively. Both values were far lower than the thresholds (95-96% for ANI and 70% for dDDH) generally accepted for new species delineation. The respiratory quinone of strain LZ-4T was Q-8. The dominant cellular fatty acids were determined as summed feature 3 (C16:1 ω6c/ω7c), summed feature 8 (C18:1 ω6c/ω7c) and C16:0. Polar lipids profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and three unidentified polar lipids. The genomic DNA G+C content of strain LZ-4T was 52.5 mol%. Based on polyphasic characterization, strain LZ-4T represents a novel species of the genus Limnobacter, for which the name Limnobacter alexandrii sp. nov. is proposed. The type strain is LZ-4T (=CCTCC AB 2019004T =KCTC 72281T).

Inhibition to crucial enzymes in the lethal effects of the dinoflagellate Karenia mikimotoi on the rotifer Brachionus plicatilis.

Blooms of the dinoflagellate Karenia mikimotoi have cause great financial losses to the marine aquaculture industry. However, the toxicity mechanism of this species is still not fully known. In this study, we evaluated the short-term effects of K. mikimotoi on the rotifer Brachionus plicatilis by micro and sub micro observing and by measuring inhibition of crucial enzymes. Behaviour disorder, mucus production, corona and cilium damage, vesical production, and body shrinkage occurred within 1 h after rotifers were treated with K. mikimotoi at a density of 3 × 104 cells/mL. Enzyme activity assays showed that K. mikimotoi at low densities significantly inhibited multiple enzymes within 3 h, and obvious density-effect trends were also observed. For instance, activity of esterase and acetylcholinesterase of rotifers significantly decreased to 94.3/83.3% and 82.8/66.9% of control treatment values in 30 and 1000 cells/mL algal treatment, respectively. Total ATPase and Na+-K+-ATPase activities of rotifers also decreased to 82.3% and 68.6% of control values in 1000 cells/mL treatment. The LDH releasement test and MDA tests showed no significant difference between algae treatment and control. It suggested that K. mikimotoi might not cause significant cytolysis and oxidative damage to rotifers, but may cause mortality by inhibiting the activity of crucial enzymes, which may lead to cell permeability disorder and body shrinkage.

Spatial and temporal disease dynamics of the parasite Hematodinium sp. in shore crabs, Carcinus maenas.

BACKGROUND: The parasitic dinoflagellates of the genus Hematodinium represent the causative agent of so-called bitter or pink crab disease in a broad range of shellfish taxa. Outbreaks of Hematodinium-associated disease can devastate local fishing and aquaculture efforts. The goal of our study was to examine the potential role of the common shore (green) crab Carcinus maenas as a reservoir for Hematodinium. Carcinus maenas is native to all shores of the UK and Ireland and the North East Atlantic but has been introduced to, and subsequently invaded waters of, the USA, South Africa and Australia. This species is notable for its capacity to harbour a range of micro- and macro-parasites, and therefore may act as a vector for disease transfer. METHODS: Over a 12-month period, we interrogated 1191 crabs across two distinct locations (intertidal pier, semi-closed dock) in Swansea Bay (Wales, UK) for the presence and severity of Hematodinium in the haemolymph, gills, hepatopancreas and surrounding waters (eDNA) using PCR-based methods, haemolymph preparations and histopathology. RESULTS: Overall, 13.6% were Hematodinium-positive via PCR and confirmed via tissue examination. Only a small difference was observed between locations with 14.4% and 12.8% infected crabs in the Dock and Pier, respectively. Binomial logistic regression models revealed seasonality (P < 0.002) and sex (P < 0.001) to be significant factors in Hematodinium detection with peak infection recorded in spring (March to May). Male crabs overall were more likely to be infected. Phylogenetic analyses of the partial ITS and 18S rRNA gene regions of Hematodinium amplified from crabs determined the causative agent to be the host generalist Hematodinium sp., which blights several valuable crustaceans in the UK alone, including edible crabs (Cancer pagurus) and langoustines (Nephrops norvegicus). CONCLUSIONS: Shore crabs were infected with the host generalist parasite Hematodinium sp. in each location tested, thereby enabling the parasite to persist in an environment shared with commercially important shellfish.

Carbonic anhydrase 2-like in the giant clam, Tridacna squamosa: characterization, localization, response to light, and possible role in the transport of inorganic carbon from the host to its symbionts.

The fluted giant clam, Tridacna squamosa, lives in symbiosis with zooxanthellae which reside extracellularly inside a tubular system. Zooxanthellae fix inorganic carbon (Ci) during insolation and donate photosynthate to the host. Carbonic anhydrases catalyze the interconversion of CO2 and HCO3-, of which carbonic anhydrase 2 (CA2) is the most ubiquitous and involved in many biological processes. This study aimed to clone a CA2 homolog (CA2-like) from the fleshy and colorful outer mantle as well as the thin and whitish inner mantle of T. squamosa, to determine its cellular and subcellular localization, and to examine the effects of light exposure on its gene and protein expression levels. The cDNA coding sequence of CA2-like from T. squamosa comprised 789 bp, encoding 263 amino acids with an estimated molecular mass of 29.6 kDa. A phenogramic analysis of the deduced CA2-like sequence denoted an animal origin. CA2-like was not detectable in the shell-facing epithelium of the inner mantle adjacent to the extrapallial fluid. Hence, CA2-like is unlikely to participate directly in light-enhanced calcification. By contrast, the outer mantle, which contains the highest density of tertiary tubules and zooxanthellae, displayed high level of CA2-like expression, and CA2-like was localized to the tubule epithelial cells. More importantly, exposure to light induced significant increases in the protein abundance of CA2-like in the outer mantle. Hence, CA2-like could probably take part in the increased supply of inorganic carbon (Ci) from the host clam to the symbiotic zooxanthellae when the latter conduct photosynthesis to fix Ci during light exposure.

X-Cells Are Globally Distributed, Genetically Divergent Fish Parasites Related to Perkinsids and Dinoflagellates.

"X-cells" have long been associated with tumor-like formations (xenomas) in marine fish, including many of commercial interest. The name was first used to refer to the large polygonal cells that were found in epidermal xenomas from flatfish from the Pacific Northwest [1]. Similar looking cells from pseudobranchial xenomas had previously been reported from cod in the Atlantic [2] and Pacific Oceans [3]. X-cell pathologies have been reported from five teleost orders: Pleuronectiformes (flatfish), Perciformes (perch-like fish), Gadiformes (cods), Siluriformes (catfish), and Salmoniformes (salmonids). Various explanations have been elicited for their etiology, including being adenomas or adenocarcinomas [4, 5], virally transformed fish cells [6-8], or products of coastal pollution [9, 10]. It was hypothesized that X-cells were protozoan parasites [1, 11-13], and although recent molecular analyses have confirmed this, they have failed to place them in any phylum [14-18], demonstrating weak phylogenetic associations with the haplosporidians [16] or the alveolates [15]. Here, we sequenced rRNA genes from European and Japanese fish that are known to develop X-cell xenomas. We also generated a metagenomic sequence library from X-cell xenomas of blue whiting and Atlantic cod and assembled 63 X-cell protein-coding genes for a eukaryote-wide phylogenomic analysis. We show that X-cells group in two highly divergent clades, robustly sister to the bivalve parasite Perkinsus. We formally describe these as Gadixcellia and Xcellia and provide a phylogenetic context to catalyze future research. We also screened Atlantic cod populations for xenomas and residual pathologies and show that X-cell infections are more prevalent and widespread than previously known.

Toxic potential of palytoxin.

This review briefly describes the origin, chemistry, molecular mechanism of action, pharmacology, toxicology, and ecotoxicology of palytoxin and its analogues. Palytoxin and its analogues are produced by marine dinoflagellates. Palytoxin is also produced by Zoanthids (i.e. Palythoa), and Cyanobacteria (Trichodesmium). Palytoxin is a very large, non-proteinaceous molecule with a complex chemical structure having both lipophilic and hydrophilic moieties. Palytoxin is one of the most potent marine toxins with an LD50 of 150 ng/kg body weight in mice exposed intravenously. Pharmacological and electrophysiological studies have demonstrated that palytoxin acts as a hemolysin and alters the function of excitable cells through multiple mechanisms of action. Palytoxin selectively binds to Na(+)/K(+)-ATPase with a Kd of 20 pM and transforms the pump into a channel permeable to monovalent cations with a single-channel conductance of 10 pS. This mechanism of action could have multiple effects on cells. Evaluation of palytoxin toxicity using various animal models revealed that palytoxin is an extremely potent neurotoxin following an intravenous, intraperitoneal, intramuscular, subcutaneous or intratracheal route of exposure. Palytoxin also causes non-lethal, yet serious toxic effects following dermal or ocular exposure. Most incidents of palytoxin poisoning have manifested after oral intake of contaminated seafood. Poisonings in humans have also been noted after inhalation, cutaneous/systemic exposures with direct contact of aerosolized seawater during Ostreopsis blooms and/or through maintaining aquaria containing Cnidarian zoanthids. Palytoxin has a strong potential for toxicity in humans and animals, and currently this toxin is of great concern worldwide.

Mode of action of membrane-disruptive lytic compounds from the marine dinoflagellate Alexandrium tamarense.

Certain allelochemicals of the marine dinoflagellate Alexandrium tamarense cause lysis of a broad spectrum of target protist cells but the lytic mechanism is poorly defined. We first hypothesized that membrane sterols serve as molecular targets of these lytic compounds, and that differences in sterol composition among donor and target cells may cause insensitivity of Alexandrium and sensitivity of targets to lytic compounds. We investigated Ca(2+) influx after application of lytic fractions to a model cell line PC12 derived from a pheochromocytoma of the rat adrenal medulla to establish how the lytic compounds affect ion flux associated with lysis of target membranes. The lytic compounds increased permeability of the cell membrane for Ca(2+) ions even during blockade of Ca(2+) channels with cadmium. Results of a liposome assay suggested that the lytic compounds did not lyse such target membranes non-specifically by means of detergent-like activity. Analysis of sterol composition of isolates of A. tamarense and of five target protistan species showed that both lytic and non-lytic A. tamarense strains contain cholesterol and dinosterol as major sterols, whereas none of the other tested species contain dinosterol. Adding sterols and phosphatidylcholine to a lysis bioassay with the cryptophyte Rhodomonas salina for evaluation of competitive binding indicated that the lytic compounds possessed apparent high affinity for free sterols and phosphatidylcholine. Lysis of protistan target cells was dose-dependently reduced by adding various sterols or phosphatidylcholine. For three tested sterols, the lytic compounds showed highest affinity towards cholesterol followed by ergosterol and brassicasterol. Cholesterol comprised a higher percentage of total sterols in plasma membrane fractions of A. tamarense than in corresponding whole cell fractions. We conclude therefore that although the molecular targets of the lytic compounds are likely to involve sterol components of membranes, A. tamarense must have a complex self-protective mechanism that still needs to be addressed.

Physiological responses of three crustacean species to infection by the dinoflagellate-like protist Hematodinium (Alveolata: Syndinea).

This is the first study comparing physiological responses of three decapod species to infection by parasites of the genus Hematodinium, which belongs to the dinoflagellate-like Syndinea. Responses varied profoundly between the crabs Carcinus maenas and Cancer pagurus (Brachyura), but also differed to those of hermit crabs, Pagurus bernhardus (Anomura). Osmoregulatory capacity was reduced significantly in Hematodinium-infected C. maenas, haemolymph pH increased in parasitised C. pagurus and P. bernhardus, and L-lactate concentration decreased in infected P. bernhardus. Changes to tissues and exoskeletons were observed in C. pagurus, but not in C. maenas and P. bernhardus.

Detection and quantification of maitotoxin-like compounds using a neuroblastoma (Neuro-2a) cell based assay. Application to the screening of maitotoxin-like compounds in Gambierdiscus spp.

SK&F 96365 was used in a Neuroblastoma (Neuro-2a) cell based assay to determine the production of maitotoxin-like (MTX-like) compounds in two strains of Gambierdiscus spp. A 2.5 hour assay was effective for the detection of the MTX-induced toxic effects with a concentration that inhibited 50% cell viability (IC(50)) equivalent to 3.38 nM MTX. Evidence was found for the production of MTX-like compounds in both Gambierdiscus strains studied at concentrations of 404 and 36.7 nmoles MTX equivalence per 10(6) cells. The assay is proposed as an efficient approach to the detection and quantification of MTX-like compounds in Gambierdiscus spp.

Cell-based assay coupled with chromatographic fractioning: a strategy for marine toxins detection in natural samples.

Cell-based assays (CBA) have been proposed for the evaluation of toxicity caused by marine toxins in natural samples (fish, shellfish and microalgae). However, their application has been hindered due to the interferences present in biological matrices that may cause cellular response and interfere in toxicity evaluation. This work reviews in an extensive introduction the use of CBA for toxicity evaluation of marine toxins. Afterwards, the coupling of chromatographic fractioning with neuroblastoma Neuro-2a CBA is presented to enhance the applicability of CBA for complex matrices. Examples of application are provided for mussel samples (Mytilus galloprovincialis) and microalgae (Gambierdiscus sp.), and the results demonstrated the great potential of the combined strategy for reliable toxicological evaluation without ethical concern. Fractioning of an equivalent of 72 mg eq mL(-1) of mussel sample allowed the identification of non-toxic and toxic fractions whereas only 2.5mg eq mL(-1) of non-purified mussel sample was responsible for 20% of cell mortality. Furthermore, the application of CBA allowed selectively distinguishing between ciguatoxin-like and other unspecific toxicity in Gambierdiscus sp. extract.

Monitoring of brevetoxins in the Karenia brevis bloom-exposed Eastern oyster (Crassostrea virginica).

Brevetoxin uptake and elimination were examined in Eastern oyster (Crassostrea virginica) exposed to recurring blooms of the marine alga Karenia brevis in Sarasota Bay, FL, over a three-year period. Brevetoxins were monitored by in vitro assays (ELISA, cytotoxicity assay, and receptor binding assay) and LC-MS, with in vivo toxicity of shellfish extracts assessed by the traditional mouse bioassay. Measurements by all methods reflected well the progression and magnitude of the blooms. Highest levels recorded by mouse bioassay at bloom peak were 157 MU/100g. Oysters were toxic by mouse bioassay at levels >or=20 MU/100g for up to two weeks after bloom dissipation, whereas brevetoxins were measurable by in vitro assays and LC-MS for several months afterwards. For the structure-based methods, summed values for the principal brevetoxin metabolites of PbTx-2 (cysteine and cysteine sulfoxide conjugates), as determined by LC-MS, were highly correlated (r(2)=0.90) with composite toxin measurements by ELISA. ELISA and LC-MS values also correlated well (r(2)=0.74 and 0.73, respectively) with those of mouse bioassay. Pharmacology-based cytotoxicity and receptor binding assays did not correlate as well (r(2)=0.65), and were weakly correlated with mouse bioassay (r(2)=0.48 and 0.50, respectively). ELISA and LC-MS methods offer rapid screening and confirmation, respectively, of brevetoxin contamination in the oyster, and are excellent alternatives to mouse bioassay for assessing oyster toxicity following K. brevis blooms.

Conservation in the first internal transcribed spacer region (ITS1) in Hematodinium species infecting crustacean hosts found in the UK and Newfoundland.

Parasitic dinoflagellates in the genus Hematodinium infect a number of decapod crustaceans in waters off the UK, including the Norway lobster Nephrops norvegicus and the edible crab Cancer pagurus. This study investigated sequence variability in the first internal transcribed spacer (ITS1) region of the ribosomal RNA complex of Hematodinium spp. infecting N. norvegicus, C. pagurus, and Pagurus bernhardus from 4 locations in the UK and from the Hematodinium sp. infecting Chionoecetes opilio from the province of Newfoundland and Labrador, Canada. Phylogenetic analysis of the Hematodinium ITS1 sequences from N. norvegicus, C. pagurus, P. bernhardus and C. opilio suggest that these crustaceans are infected with the same species of Hematodinium. Length variability of the ITS1 region was observed (324 to 345 bp) and attributed to 4 variable microsatellite regions (CATG)n' (GCC)nTCCGC(TG)n' (TA)n' and (GAA)n(GGA)n within the sequenced ITS1 fragment. The observed variation may be due to co-infection of the host crustacean with several different strains of Hematodinium or differences among copies of ITS1 region within the genome of a single parasite cell. The Hematodinium ITS1 sequence from N. norvegicus, C. pagurus, P. bernhardus and C. opilio isolates was sufficiently conserved in primer binding regions targeted by previous molecular diagnostic assays; therefore, we suggest that this assay could be used to screen for Hematodinium infections in these crustacean hosts.

Characterization of polar brevetoxin derivatives isolated from Karenia brevis cultures and natural blooms.

Several novel brevetoxin derivatives were isolated and identified in Karenia brevis cultures and natural blooms by using solid phase extraction (SPE) and LC/MS(MS) techniques. These analogs were more polar compared with previously described brevetoxins, and were poorly extractable by conventional non-polar solvent (chloroform) partitioning. Brevetoxin analogs were structurally confirmed as hydrolyzed (open A-ring) forms of brevetoxins PbTx-1, PbTx-7, PbTx-2, and PbTx-3, and of oxidized PbTx-1 and PbTx-2. Some of these open A-ring derivatives were in greater abundance than their non-hydrolyzed counterparts. All were in much greater abundance in bloom water filtrate compared with cell-rich fractions. Open A-ring compounds were cytotoxic in mouse neuroblastoma (N2a) cell assay. In the K. brevis bloom-exposed Eastern oyster, brevetoxin metabolites with opened A rings were identified (e.g., open-ring cysteine-PbTx conjugates), contributing to their overall toxin burden.

Cytotoxicity in the marine dinoflagellate Prorocentrum mexicanum from Brazil.

The microscopic algae in the oceans are crucial food for filter feeding bivalve shellfish (oysters, mussels, scallops, clams, etc.) as well as for the larvae of commercially important crustaceans. Some species of microalgae have the capacity to produce potent toxins, such as saxitoxins and ciguatoxins, which may intoxicate humans. Among the marine phytoplankton, the dinoflagellates are the main toxin producers. Studies on the marine phytoplankton from the São Sebastião Channel, southeastern coast of Brazil, showed a great diversity of dinoflagellates. Some species were collected and cultured at the Marine Biology Center of the São Paulo University (USP). The polar (PEs) (aqueous) and apolar (AEs) (methylene chloride) extracts of the cultivated dinoflagellate species were tested on different stages of the sea urchin development, on mouse erythrocytes and on microfilaments organization in a neuroblastoma cell line. Prorocentrum mexicanum PE and AE induced cells anomalies and cell division inhibition of sea urchin eggs at EC50 of 78.75 microg/mL (95% CI from 32.56 to 190.50) and 22.50 microg/mL (95% CI from 2.96 to 170.80) respectively (n=3). Both AE and PE of P. mexicanum induced hemolysis with EC50 of 65.07 microg/mL (95% CI from 27.40 to 154.60) and 84.29 microg/mL (95% CI from 53.26 to 133.40 microg/mL), respectively. P. mexicanum PE was tested in immunofluorescence for actin filaments organization in neuroblastoma cultured cell.

Occupational exposure to aerosolized brevetoxins during Florida red tide events: effects on a healthy worker population.

Karenia brevis (formerly Gymnodinium breve) is a marine dinoflagellate responsible for red tides that form in the Gulf of Mexico. K. brevis produces brevetoxins, the potent toxins that cause neurotoxic shellfish poisoning. There is also limited information describing human health effects from environmental exposures to brevetoxins. Our objective was to examine the impact of inhaling aerosolized brevetoxins during red tide events on self-reported symptoms and pulmonary function. We recruited a group of 28 healthy lifeguards who are occupationally exposed to red tide toxins during their daily work-related activities. They performed spirometry tests and reported symptoms before and after their 8-hr shifts during a time when there was no red tide (unexposed period) and again when there was a red tide (exposed period). We also examined how mild exercise affected the reported symptoms and spirometry tests during unexposed and exposed periods with a subgroup of the same lifeguards. Environmental sampling (K. brevis cell concentrations in seawater and brevetoxin concentrations in seawater and air) was used to confirm unexposed/exposed status. Compared with unexposed periods, the group of lifeguards reported more upper respiratory symptoms during the exposed periods. We did not observe any impact of exposure to aerosolized brevetoxins, with or without mild exercise, on pulmonary function.

La marea roja causada por el dinoflagelado Alexandrium tamarense en la costa Pacífica colombiana (2001) / The red tide caused by the dinoflagellate Alexandrium tamarense in the Colombian Pacific coast (2001)

El 26 de marzo de 2001 se registró por primera vez en las aguas del Pacífico colombiano, en el área de la Ensenada de Tumaco, la aparición de una marea roja producida por la especie Alexandrium tamarense con valores que superaron las 7.5 x 106 céls l-1 , la marea se mantuvo hacia la región oceánica, derivando en sentido sur-norte por efecto de las corrientes, hasta ser vista por ultima vez cerca a la Isla Gorgona, casi tres semanas después. Un año mas tarde, en marzo de 2002, una segunda proliferación, conformada por la misma especie, se reportó cerca de Cabo Corrientes con una concentración de 1.6 x 106 céls l -1 . Ambos casos estuvieron relacionados con la presencia de bajas temperaturas en la superficie del mar y no estuvieron asociadas con fenómenos de intoxicación y mortalidad

From April 26th to May 15th 2001, a large algae bloom was observed off Tumaco Bay on the Pacific coast of Colombia. This was the first harmful algae bloom (HAB) reported in the region, and reached Gorgona Island, about 120 km north. Ayear later, starting March 2002, an offshore HAB developed from Cabo Corrientes North to Solano Bay. The typical abundance during the blooms reached 7.5 x 106 cells l-1 for the 2001 event and 1.6 x 106 cells l-1 for the 2002 event. During both events, low temperature and high salinity were recorded. Typical measurements in the area are 27-27.5°C and 30-31.5 psu. Values observed during the two events were 24-24.6°C and 33-34 psu; 3°C below normal and more than 2.5 psu above average values. These conditions are indicative of local upwelling processes at the time of the events. On both occasions, cells corresponding to the Alexandrium catenella/fundeyense/tamarense complex represented 99-100% of the biomass. It was difficult to differentiate the cells from A. catenella, but the presence of short chains of only 4 cells (single cells represented most of the biomass) was suggestive of A. tamarense. Shape, dimensions, and detailed structure of the apical pore complex, first apical plate, posterior sulcal plate, and position of the ventral pore on plate 1’ of cells were consistent with the description of A. tamarense, which has not been reported in the tropical East Pacific. The Control Center of Pacific Contamination of the Maritime General Direction of the Colombian Navy has been monitoring the area since 1994 without finding this species or HABs. This leads us to consider the two events as caused by recently introduced species, where local upwelling processes favor permanent and cyclic HABs. However, during these two events, there were no reports of effects on marine biota or of human poisoning, probably because the blooms occurred some distance offshore and far from exploited shellfish beds

Presencia de los dinoflagelados Ceratium dens, C. fusus y C. furca (Gonyaulacales: Ceratiaceae) en el Golfo de Nicoya, Costa Rica / Presence of the dinoflagellates Ceratium dens, C. fusus and C. furca (Gonyaulacales: Ceratiaceae) in Golfo de Nicoya, Costa Rica

Las proliferaciones algales nocivas (PAN) son un fenómeno frecuente en el Golfo de Nicoya. La morfología y fisiología de estas microalgas son muy importantes debido al hecho de que las especies involucradas en estos eventos presentan características adaptativas. PAN producidas por dinoflagelados del género Ceratium son comunes en algunas partes del mundo, sin embargo, a pesar del hecho de que la producción de altas concentraciones de toxinas paralíticas sólo se ha documentado a nivel experimental, éste género ha sido asociado con la mortalidad de organismos acuáticos y con la disminución en la calidad del agua, así como el responsable de la mortalidad de ostras y larvas de gambas. Algunas especies del género Ceratium han sido asociados con eventos de mortalidad en peces. Durante un reciente fenómeno PAN que ocurrió cerca de la Isla Tortuga en el Golfo de Nicoya, los pescadores en el área informaron muertes masivas de peces enjaulados. Se tomaron muestras de una PAN que producía una discoloración anaranjada en el agua y tenía un olor fétido. Estas muestras fueron procesadas para observar el detalle ultraestructural usando el microscopio electrónico. En muestras tomadas en superficie, los dinoflagelados observados fueron Ceratium furca, C. fusus y C. dens. El conteo celular reveló cuatro millones de células de este género por litro. Los cambios morfológicos de estas especies son muy variables; por consiguiente la microscopía electrónica es una herramienta muy útil en el estudio ultraestructural de estos organismos. Esta es la primera vez que tres especies del género Ceratium se conocen como productoras concurrentes de PAN en Costa Rica

Harmful Algae Blooms (HAB) are a frequent phenomenon in the Gulf of Nicoya, Costa Rica, as in other parts of the world. The morphology and physiology of these microalgae are important because HAB species have adaptive characteristics. The production of high concentrations of paralytic toxins by Ceratium dinoflagellates has only been documented at the experimental level. However, this genus has been associated with the mortality of aquatic organisms, including oyster and shrimp larva, and fish, and with decreased water quality. Recently, fishermen reported massive mortality of encaged fish near Tortuga Island (Gulf of Nicoya). Samples were taken from an algal bloom that had produced an orange coloration and had a strong foul-smelling odor. Ultrastructural details were examined with scanning electron microscopy. The dinoflagellates Ceratium dens, C. furca and C. fusus were found in samples taken at the surface. The cell count revealed four million cells of this genus per liter. The morphological variability of these species is high; therefore electron microscopy is an useful tool in the ultrastructural study of these organisms. This is the first time that three Ceratium species are reported concurrently producing harmful blooms in Costa Rica

Proliferaciones algales nocivas de cianobacterias (Oscillatoriaceae) y dinoflagelados (Gymnodiniaceae) en el Golfo de Nicoya, Costa Rica / Harmful blooms of cyanobacteria (Oscillatoriaceae) and dinoflagellates (Gymnodiniaceae) in the Golfo de Nicoya, Costa Rica

Recently, the Pacific coast of Costa Rica has experienced an increase in both magnitude and frequency of harmful algae blooms (HAB). The lack of data regarding the dynamics of these events in the area, and the species of microalgae that produce them, are themes of great interest. The blooms have produced negative impacts on fishery resources and on human health in Costa Rica. In May 2002 a HAB left a large number of dead fish along the central Pacific coast. Water samples were collected using a phytoplankton net and fixed for subsequent processing by electron microscopy. In addition, a one liter sample of surface water was taken for later cell count. In the observed HAB, the dominating organisms found were the cyanobacteria Trichodesmiun erythraeum surrounded by high concentrations of Gram - bacteria and the dinoflagellate Cochlodinium cf. polykrikoides. T. erythraeum, is one of the most important N 2 fixing cyanobacteria in marine waters that has been associated with HAB events in diverse parts of the world as well as with symptoms that produce contact dermatitis and other discomforts. C. cf. polykrikoides is a dinoflagellete associated with fish kills; although the type of associated toxins are unknown. In a national newspaper 17 cases of intoxication in humans were reported during this same period, which presented respiratory disorders and burning of the eyes. This is the first report in Costa Rica where a cyanobacteria and a dinoflagellate were observed together producing HAB

Recientemente, la costa del Pacífico de Costa Rica ha experimentado un aumento en magnitud y frecuencia de proliferaciones algales nocivas (PAN). La falta de datos con respecto a la dinámica de estos eventos en el área, y las especies de microalgas que los producen, son temas de mucho interés. Las PAN han producido impactos negativos en los recursos pesqueros y en la salud humana en Costa Rica. En mayo de 2002, una PAN se observó frente a las playas a lo largo del Pacífico central; además, se observó un gran número y diversidad de peces muertos. Las muestras de agua fueron recolectadas usando una red de fitoplancton y se fijaron para ser procesadas posteriormente con microscopía electrónica. Asimismo, se tomó un litro de muestra de la superficie para su posterior conteo celular. En la PAN observada, los organismos dominantes fueron la cianobacteria Trichodesmiun erythraeum, rodeada por altas concentraciones de bacterias Gram -, y el dinoflagelado Cochlodinium cf. polykrikoides. T. erythraeum, es una de las principales cianobacterias fijadoras de N2 atmosférico en aguas marinas, y se ha asociado con los eventos PAN en diversas partes del mundo, produciendo síntomas como dermatitis por contacto con la brisa marina y otras afecciones. C. cf. polykrikoides es un dinoflagelado asociado con mortalidad de peces; aunque el tipo de toxinas asociadas aún es desconocido. En un periódico nacional se reportaron 17 casos de intoxicación en humanos durante este mismo periodo de PAN, los afectados presentaron desórdenes respiratorios e irritación en los ojos. Éste es el primer informe en Costa Rica dónde se observaron cianobacterias y dinoflagelados juntos produciendo PAN. Este tipo de fenómenos debe analizarse y debe documentarse, para habilitar una comprensión en la dinámica y los factores asociados con el aumento de estos eventos en el Pacífico costarricense

Paralytic shellfish toxins in the chocolata clam, Megapitaria squalida (Bivalvia: Veneridae), in Bahia de La Paz, Gulf of California

Occurrence and toxic profiles of paralytic shellfish toxins (PST) in the chocolata clam Megapitaria squalida were investigated. From December 2001 to December 2002, 25 clams were obtained monthly from Bahía de La Paz, Gulf of California. Additionally, net (20 µm) and bottle phytoplankton samples were also collected to identify toxic species. Toxins were analyzed by HPLC with post-column oxidation and fluorescence detection. Toxicity in the clam was low and varied from 0.14 to 5.46 µg/STXeq/100 g. Toxicity was detected in December, March, April, June, and August. Toxin profile was composed mainly by STX, GTX2, GTX3, dcGTX2, dcGTX3, C2, dcSTX and B1. Gymnodinium catenatum was the only PST-producing dinoflagellate identified in the phytoplankton samples throughout the study period. G. catenatum was observed mainly in net samples from December 2001 to December 2002; however, in bottle samples, G. catenatum was only observed in five months. Highest abundance (2 600 cells l-1 ) was observed in March and the lowest (160 cells l-1 ) in June. G. catenatum mainly formed two-cell chains and rarely four or eight. The presence of PST in net phytoplankton samples support the fact that G. catenatum is the main source of PST in the clams. This study represents the first report of PST toxins in the chocolata clam from Bahía de La Paz

Se investigó la ocurrencia así como los perfiles de toxinas paralíticas (PST) en la almeja chocolata Megapitaria squalida (Sowerby, 1835), de la cual se recolectaron mensualmente 25 ejemplares de diciembre del 2001 a diciembre del 2000 en La Bahía de La Paz, Golfo de California. Simultáneamente, se obtuvieron muestras de fitoplancton de botella y de red (20 µM) para identificar especies tóxicas, así como para detectar la presencia de toxinas paralíticas. Las toxinas se analizaron por HPLC con una oxidación post-columna y detección fluorescente. La toxicidad en las almejas fue baja y varió de 0.14 a 5.46 µg/STXeq/100 g y se detectó en diciembre, marzo, abril, junio y agosto. El perfil de toxinas estuvo compuesto principalmente por STX, GTX2, GTX3, dcGTX2, dcGTX3, C2, dcSTX y B1, siendo similar al perfil observado en las muestras de red. Gymnodinium catenatum fue la única especie productora de toxinas paralíticas identificada en las muestras de fitoplancton a través de todo el período del estudio. G. catenatum se observó principalmente en muestras de red correspondientes a diciembre 2001-diciembre 2002; sin embargo, en muestras de botella, G. Catenatum solo fue observado en cinco meses. La abundancia más alta (2 600 céls l-1 ) se encontró en marzo y la más baja (160 céls l-1 ) en junio. G. catenatum se presentó formando cadenas de dos células y raramente de cuatro u ocho. La presencia de toxinas en las muestras de red donde esta especie es frecuente, apoya la idea de que G. catenatum sea la fuente principal de PST en las almejas. Este estudio representa el primer reporte de toxinas paralíticas en la almeja chocolata de Bahía de La Paz

Marea roja: su situación en Chile / Red tide: its characteristics in Chile

Se conoce con el nombre de marea roja a un fenómeno de cambio de color de las aguas del mar ocasionado por el aumento de algunos organismos del plancton que se multiplican explosivamente y se concentran en la superficie de las aguas. Las especies tóxicas producen en su metabolismo compuestos químicos de muy alta toxicidad denominados toxinas marinas, que pueden ser acumuladas por moluscos filtradores en concentraciones dañinas para ser humano.De acuerdo a sus efectos estos compuestos se han clasificado como toxinas marinas paralizantes, neurotóxicas amnésicas, diarreicas y ciguatéricas. Las toxinas diarreicas y paralizantes son las encontradas con mayor frecuencia en nuestras costas.Debido a las repercusiones que la marea roja tiene para la salud del hombre y para la economía de países con grandes áreas de costa como Chile, ésta reviste especialimportancia y es objeto de constante vigilancia y estudio por parte de las autoridades marítimas y de salud. Por ello es indispensable que el médico general reconozca e inicie oportunamente el tratamiento de los cuadros de intoxicación aguda.