Florecimiento algal nocivo de Pyrodinium bahamense en diciembre 2018 en la costa del Pacífico de Guatemala / Harmful algal bloom of Pyrodinium bahamense in December 2018 in the Pacific coast of Guatemala

Los florecimientos algales nocivos (FAN) son eventos naturales que ocurren cuando una o más especies de microalgas proliferan en concentraciones que pueden causar daño a los organismos acuáticos y a seres humanos que los consuman. En las últimas décadas, se ha registrado a nivel mundial un aumento de eventos de mortalidad alarmante de organismos acuáticos e intoxicaciones en seres humanos causadas por toxinas producidas por microalgas. En Guatemala existe escasa información sobre eventos FAN, no obstante, en diciembre de 2018 ocurrió un FAN en la costa Pacífico de Guatemala. La Comisión Nacional para la Vigilancia y el Control de la Marea Roja Tóxica recolectó muestras biológicas e identificó la presencia de 3,000 cel/L del dinoflagelado Pyrodinium bahamense. El bioensayo en ratón de las muestras, indica concentraciones de saxitoxina de 8,236 UR/100 g y 6,559 UR/100 g, para los días 20 y 27 de diciembre, respectivamente. Estas concentraciones no han sido reportadas previamente en Guatemala, y pueden ser potencialmente tóxicas para la salud pública. Se recomienda mantener un monitoreo de FAN para prevenir impactos negativos en la salud pública y ambiental.

Harmful algal blooms (HABs) are natural events that occur when one or more species of microalgae proliferate at concentrations that can cause damage to aquatic organisms and to those who consume them. In Guatemala very little information exists on HAB events, although in December 2018 has occurred a HABs at the Pacific Coast of Guatemala. A biological sample were collected by the National Commission for Surveillance and Control of Toxic Red Tide. The samples showed 3,000 cel/L of the dinoflagellate Pyrodinium bahamense. During the bioassay for saxitoxin, the concentrations reached 8,236 MU/100 g and 6,559 MU/100 g, for December 20th and 27th, respectively. These concentrations has not been reported previously for Guatemala, and could be a risk in the public health. This result emphasizes the importance of maintaining the HABS monitoring program to prevent negative impact on public environmental health.

Proposta de modelo de exposição humana à saxitoxina em águas de recreação e de abastecimento público do reservatório Itupararanga / Proposal of a model of human exposure to saxitoxin in recreational waters and public supply of the Itupararanga reservoir

O aporte artificial de nutrientes nos ambientes aquáticos tem favorecido a incidência de florações de cianobactérias, causando impactos na qualidade da água e colocando em risco a saúde humana. O gênero de cianobactéria Cylindrospermopsis, amplamente distribuído em água doce pelo mundo, é capaz de produzir diversas cianotoxinas, dentre elas saxitoxina e análogos. O objetivo deste trabalho foi elaborar um modelo de exposição referente à ingestão de água contaminada por saxitoxina e análogos através da recreação de contato primário e do consumo de água. Os dados de densidade de células de Cylindrospermopsis e de concentrações de saxitoxina e análogos na água bruta foram obtidos da Rede de Monitoramento de Qualidade de Águas Superficiais da CETESB (2015 e 2016) do reservatório Itupararanga. O modelo conceitual de exposição e o cálculo da ingestão diária crônica de saxitoxina e análogos consideraram: concentrações máxima e mínima de saxitoxina e análogos encontradas na água bruta do reservatório, vias de exposição, população exposta, taxa de ingestão de água, frequência e duração da exposição, peso corporal para adultos (maiores de 21 anos) e crianças (menores de 5 anos), para cenários de exposição relativos à recreação de contato primário e consumo de água potável. Os valores de células de cianobactérias estiveram entre 2.216 e 203.082 céls.mL-1 e as concentrações de saxitoxina e análogos estiveram entre 0,04 e 0,37 µg equivalente STX..L-1. Os resultados obtidos na análise da água bruta do reservatório, estiveram abaixo do valor estabelecido na legislação brasileira para água potável (3 µg equivalente STX.L-1), mas não atenderiam as especificações adotadas por Oregon e Ohio (0,3 µg equivalentes STX.L-1) e à proposta em discussão no Comitê Permanente para Gestão Integrada da Qualidade da Água destinada ao Consumo Humano no Estado de São Paulo (0,13 µg equivalente STX.L-1), indicando a possibilidade da ocorrência de efeito adverso à saúde em grupos sensíveis que utilizem água do reservatório, dependendo da eficiência de remoção dessas cianotoxinas no processo de tratamento. Os valores de ingestão diária crônica estimados, para recreação, variaram de 2,55 x 10-6 a 1,08 x 10-4 µg equivalente STX. kg-1.dia-1 (adultos) e 2,78 x 10-9 a 4,35 x 10-4 µg equivalente STX. kg-1.dia-1 (crianças). Para ingestão de água potável, esses valores variaram de 2,5 x 10-4 a 1,47 x 10-2 µg equivalente STX. kg-1.dia-1 (adultos) e 5,36 x 10-5 a 2,55 x 10-2 µg equivalente STX. kg-1.dia-1 (crianças). Esses valores indicaram que as crianças estão mais expostas que os adultos, fato preocupante, pois elas são mais sensíveis aos efeitos de substâncias tóxicas, e apesar de estudos indicarem efeitos da saxitoxina em cultura de células neurais, sua capacidade de atravessar a barreira hematoencefálica e de causar danos cognitivos em ratos, os efeitos da exposição crônica em humanos ainda são desconhecidos. Não foi possível estimar os riscos à saúde da população exposta à saxitoxina e análogos, pela falta de dados na literatura, porém o modelo de exposição proposto é um avanço para a identificação de lacunas para a condução da avaliação de risco à saúde humana considerando a exposição crônica

The artificial enrichment of nutrients in aquatic environments has favored the incidence of cyanobacteria blooms, causing impacts on water quality and putting human health at risk. The genus of cyanobacteria Cylindrospermopsis, widely distributed throughout the world in freshwater environments, is able to produce cianotoxin, among them saxitoxin and its analogues. The objective of this work was to elaborate a model of exposure referring to the ingestion of water contaminated by saxitoxin and analogues through recreation of primary contact and water consumption. Cylindrospermopsis cell density and saxitoxin and analogues concentrations in surface water were obtained from the CETESB Surface Water Quality Monitoring Network for the Itupararanga reservoir in 2015 and 2016. The development of the conceptual model of exposure and the calculation of the chronic daily intake of saxitoxin and analogues, considered: maximum and minimum concentrations of saxitoxin and analogues found in the raw water of the reservoir, route of exposure, exposed population, rate of water intake, frequency and duration of exposure, body weight for adults (over 21 years) and children (under 5 years), for exposures scenarios concerning recreation of primary contact and consumption of drinking water. The values of cyanobacteria cells were between 2,216 and 203,082 cells.L-1 and the concentrations of saxitoxin and analogues were between 0.04 and 0.37 µg STX equivalent. L-1. The comparison of these variables indicated a significant positive correlation between them. The results obtained in the raw water analysis of the reservoir were below the value established in Brazilian legislation for drinking water (3 µg equivalent STX.L-1), but would not meet the specifications adopted by US states Oregon and Ohio (0.3 µg equivalents.L-1) and the proposed value for the Standing Committee for Integrated Water Quality Management for Human Consumption in the State of São Paulo (0,13 µg equivalent STX.L-1), indicating the possibility of adverse health effects in sensitive groups using water from the reservoir. Estimated chronic daily intake values, for recreation, ranged from 2.55 x 10-6 to 1.08 x 10-4 µg STX equivalent. kg-1.day-1 (adults) and 2.78 x 10-9 to 4.35 x 10-4 µg STX equivalent. kg-1.day-1 (children). For water intake, these values ranged from 2.5 x 10-4 to 1.47 x 10-2 µg STX equivalent. kg-1 day-1 (adults) and 5.36 x 10-5 to 2.55 x 10-2 µg STX equivalent. kg-1. day-1 (children). These values indicated that children are more exposed than adults, a worrying fact because they are generally more sensitive to the effects of toxic substances than adults, and although studies indicate that saxitoxin was able to cross the blood brain barrier and to cause cognitive damage in rats, as well as changes in neural cell cultures, the effects of chronic exposure in humans are still unknown. It was not possible to estimate the health risks of the population exposed to saxitoxin and the like, due to the lack of data in the literature, but the proposed exposure model is an advance for the identification of gaps for conducting the human health risk assessment considering the exposure chronic

Behavioral alterations induced by repeated saxitoxin exposure in drinking water

Background: Blooms of the saxitoxin-producing cyanobacterium Cylindrospermopsis raciborskii have been contaminating drinking water reservoirs in Brazil for many years. Although acute effects of saxitoxin intoxication are well known, chronic deleterious outcomes caused by repeated saxitoxin exposure still require further investigation. The aim of the present work is to investigate the effects of consumption of drinking water contaminated with C. raciborskii for 30 days on learning and memory processes in rats. Methods: The effects of saxitoxin (3 or 9 µg/L STX equivalents) or cyanobacteria on behavior was determined using the open field habituation task, elevated plus maze anxiety model task, inhibitory avoidance task, and referential Morris water maze task. Results: No effects of saxitoxin consumption was observed on anxiety and motor exploratory parameters in the elevated plus maze and open field habituation tasks, respectively. However, groups treated with 9 µg/L STX equivalents displayed a decreased memory performance in the inhibitory avoidance and Morris water maze tasks. Conclusions: These results suggest an amnesic effect of saxitoxin on aversive and spatial memories.(AU)

Behavioral alterations induced by repeated saxitoxin exposure in drinking water

Blooms of the saxitoxin-producing cyanobacterium Cylindrospermopsis raciborskii have been contaminating drinking water reservoirs in Brazil for many years. Although acute effects of saxitoxin intoxication are well known, chronic deleterious outcomes caused by repeated saxitoxin exposure still require further investigation. The aim of the present work is to investigate the effects of consumption of drinking water contaminated with C. raciborskii for 30 days on learning and memory processes in rats. Methods The effects of saxitoxin (3 or 9 g/L STX equivalents) or cyanobacteria on behavior was determined using the open field habituation task, elevated plus maze anxiety model task, inhibitory avoidance task, and referential Morris water maze task. Results No effects of saxitoxin consumption was observed on anxiety and motor exploratory parameters in the elevated plus maze and open field habituation tasks, respectively. However, groups treated with 9 g/L STX equivalents displayed a decreased memory performance in the inhibitory avoidance and Morris water maze tasks. Conclusions These results suggest an amnesic effect of saxitoxin on aversive and spatial memories.

Effects of Cylindrospermopsis raciborskii strains (Woloszynska, 1912) Senayya & Subba Raju on the mobility of Daphnia laevis (Cladocera, Daphniidae) / Efeitos de cepas de Cylindrospermopsis raciborskii (Woloszynska, 1912), Senayya & Subba Raju na mobilidade de Daphnia laevis (Cladocera, Daphniidae)

Cylindrospermopsis raciborskii is a cyanobacterium distributed worldwide that is known to produce cyanotoxins. Some of the Brazilian strains can produce saxitoxins (STXs), which are classified as neurotoxins and can paralyze cladocerans .Daphnia laevis is a cladoceran with a wide distribution in the Americas and has been studied as a possible test-organism in toxicity bioassays. The present work tested the acute effect on D laevis mobility when fed a saxitoxin-producing (STX and neoSTX) C. raciborskii strain, CYRF-01, and compared the results with the effects of a non-toxic strain (NPCS-1). Neonates (6-24 hours after birth) were exposed to concentrations of C. raciborskii varying from 102 to 106 cells·mL–1 of each strain for up to three hours. The cladocerans were then transferred to a medium without toxic filaments for 24 hours. Only the organisms exposed to the STX-producing strain showed signs of the immobilization of swimming movements, confirming the effects of the toxins. There was a linear correlation between the time required to induce stopping the swimming movement, with a shorter time to needed to induce immobilization at a higher the concentration; this correlation was inverse to the time required to recover the swimming movements (longer at higher concentrations, p < 0.1). D. laevis is a tropical and subtropical species with great potential for use in toxicity tests for the detection of STXs, despite being native to and found in a great array of freshwater bodies. This is the first assay testing STX-producing and non-producing C. raciborskii strains on D. laevis, species that are both found in Brazilian ecosystems.

Cylindrospermopsis raciborskii é uma espécie de cianobactéria, difundida mundialmente, conhecida como produtora de cianotoxinas. Algumas linhagens brasileiras são conhecidas como produtoras de saxitoxinas (STXs), as quais são classificadas como neurotoxinas e podem induzir a paralização dos movimentos natatórios em cladóceros. Daphnia laevis é um cladócero de ampla distribuição nas Américas e tem sido estudado como um organismo teste para uso em ensaios de toxicidade. Neste sentido, o presente trabalho avaliou os efeitos agudos de C. raciborskii, de uma cepa produtora de STXs (STX e neoSTX) - CYRF-01 e outra não produtora de cianotoxinas (NPCS-1) sobre a mobilidade de D. laevis. Para tanto, neonatas entre 6-24 horas de idade foram submetidas a concentrações de C. raciborskii variando de 102 a 106 céls.mL–1 de ambas as cepas, por um período de 3 horas, e transferidos posteriormente para um meio isento de filamentos tóxicos por 24horas. Apenas os organismos expostos à cepa produtora de STXs apresentaram paralisia dos movimentos natatórios, confirmando o efeito da cianotoxina. Houve uma correlação linear entre o tempo de paralização e as concentrações, isto é, nas concentrações mais altas, os organismos paralisaram num curto período de tempo, e conseqüentemente, levaram mais tempo para se recuperar (p < 0,1). D.laevis é uma espécie de região tropical e subtropical com potencial uso em bioensaios para detecção de STXs. Trata-se dos primeiros relatos acerca da resposta de D.laevis quando submetida às cepas de C. raciborskii produtora e não produtora de STXs, ambas isoladas de ecossistemas brasileiros.

Use of sodium transfer tissue biosensor (STTB) for monitoring of marine toxic organism.

A highly sensitive sodium (Na+) transfer tissue biosensor (STTB) was designed using a frog bladder membrane to measure paralytic shellfish poisons (PSP). The STTB consists, of a Na+ electrode covered by the membrane, which was then integrated into a flow-through system for continuous measurements. In the absence of Na+ channel blocker, active transfer of Na+ occurred from inside to outside across the frog membrane. When the STTB was used to measure the Na+ -dependent dissociation of PSP, it was able to detect PSB at a level contained in a single cell. However, 5 fg or higher (100 cells or more) is needed for accurate and reproducible measurements. The toxicity obtained by the STTB was significantly correlated (r = 0.9449) to that determined by the HPLC. Therefore, the simple method of the STTB can be used not only to detect a low level PSP in toxic plankton populations, but also to monitor poisons in shellfish.

Programa de Formación de Epidemiología de Campo: período 2002-2003

Contiene: estudios de brotes epidémicos, evaluación de un sistema de vigilancia, publicaciones e informes, conferencias científicas.

Paralytic Shellfish Poisoning (PSP) in Margarita Island, Venezuela

A severe outbreak of Paralytic Shellfish Poisoning (PSP) occurred in Manzanillo and Guayacán, northwestern coast of Margarita Island, Venezuela, between August and October 1991. A bloom of dinoflagellates including Prorocentrum gracile, Gymnodinium catenatum and Alexandrium tamarense seemed to be responsible for this outbreak. Levels of PSP toxins in mussels (Perna perna) exceeded the international safety limit of saxitoxin, 80 µg STX/100 g meat. PSP toxin values varied between 2 548 and 115 µg STX/100 g meat in Manzanillo, and between 1 422 and 86 µg STX/100 g meat in Guayacán. At both locations, the highest levels were detected in August, when 24 patients exhibited typical symptoms of PSP toxicity after consuming cooked mussels (16 required hospitalization). A high pressure liquid chromatographic (HPLC) procedure was recently used on the 1991 samples. The major toxin detected in samples of both locations was decarbamoyl saxitoxin (dcSTX), but low concentrations of saxitoxin were also found in Manzanillo samples. Gonyautoxins GTX1, GTX2 and GTX3 were detected only at Guayacán, while in both locations, decarbamoylgonyatouxin (dcGTX2,3) toxins were detected. These findings represent the first time that causative toxins of PSP in Venezuela have been chemically identified, and confirm the presence of dcSTX and dcGTX in mussels from the Caribbean Sea. The presence of dcSTX and dcGTX in shellfish is indicative that Gymnodinium catenatum was a causative organism for outbreak of PSP

Un severo brote de intoxicación paralizante por moluscos (PSP en inglés) ocurrió en Manzanillo y Guayacán en la costa noroeste de la Isla de Margarita, Venezuela entre agosto y octubre de 1991. Una proliferación de Prorocentrum gracile, Gymnodinium catenatum y Alexandrium tamarense causó el brote. Los niveles de PSP en mejillón (Perna perna) superaron los niveles máximos permisibles de saxitoxina, 80 µg STX/100g carne. Los niveles de toxinas variaron entre 2 548 y 115 µg STX/100 g carne en Manzanillo y entre 1 422 y 86 µg STX/100g carne en Guayacán. En ambas localidades, los máximos niveles se detectaron en agosto, cuando 24 personas presentaron síntomas típicos de PSP después de consumir mejillones cocidos (16 fueron hospitalizados). Se aplicó recientemente cromatografía líquida de alta presión (HPLC) a muestras del año 1991 y la toxina más detectada fue decarbamoyl saxitoxina (dcSTX), pero también se encontró saxitoxinas en muestras de Manzanillo. Las gonyautoxinas GTX1, GTX2 y GTX3 solo se encontraron en Guayacán; en ambas localidades se detectó decarbamoylgonyatouxin (dcGTX2,3). Estos hallazgos representan la primera vez que las toxinas causantes de un brote de PSP en Venezuela han sido químicamente identificadas, confirmando la presencia de dcSTX y dcGTX en mejillones del mar Caribe. La presencia de dcSTX y dcGTX en moluscos, indica que G. catenatum fue el organismo responsable de la intoxicación

Dinámica espacio-temporal de organismos precursores de marea roja en la costa Pacífica de América del Norte y Centroamérica / Spatial-temporal dynamics of red tide precursor organisms at the Pacific coast of North and Central America

La región costera del Pacífico de Centro y Norte America ha sido afectada de manera frecuente por fenómenos que se denominan proliferaciones algales nocivas o en inglés, "Harmful Algal Blooms" (HAB). A pesar de la gran cantidad de efectos en la salud pública, las actividades económicas y el medio ambiente, aún existe una abrumadora carencia de información en el tema. Esto no permite establecer si las causas principales de este evidente aumento en el número e intensidad de los eventos son naturales o antropogénicas. La incrementada utilización de las zonas costeras para actividades humanas encuentra paralelo en el denominado cambio climático, lo que dificulta discriminar entre la posible influencia de estos dos fenómenos. Series de datos de larga duración en regiones reducidas, así como las observaciones esporádicas realizadas cuando se presenta algún evento, nos permite comparar regímenes climáticos, condiciones geográficas, disponibilidad de nutrientes (incluyendo la eutroficación) y parámetros oceanográficos que promueven o permiten el desarrollo y mantenimiento de las proliferaciones, haciendo evidentes cambios drásticos en la biodiversidad y la biogeografía de los organismos productores de HAB

The Pacific coast of Central and North America has long been and still is impacted by the flourishing of microalgal populations known as Harmful Algal Blooms (HABs). The organisms that have caused recent HABs episodes in the region are among others, Gymnodinium catenatum, Pyrodinium bahamense var. compressum, and recently Cochlodinium cf. catenatum. In spite of the accumulated effects on the human health, the economic activities and the environment, scarce information is available on the subject. The augmented use of coastal zones for human activities is also paralleled by increased awareness of global climate changes. Thus, it is not an easy task to discriminate anthropogenic or natural phenomena, or both, as the major driving forces. The long-term data sets available for limited regions, as well as some sporadic observations during notorious blooms, allowed us to discriminate major changes in the biodiversity and biogeography of HAB organisms. Main changes refer to number of events, covered area, duration and frequency, number of blooming species and appearance of not previously reported harmful taxa. The variables more clearly related to these dynamic phenomena, seems to be sea surface temperature and wind force, but it is not yet possible to weight their contributions. The participation of rain is not fully evaluated to date. The collaborative communication among small-budget monitoring operations in the region allowed to "pass the voice" about peaking concentrations of HAB organisms, diminishing the risk of poisoning

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

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

The Gonyautoxin 2/3 epimers reduces anal tone when injected in the anal sphincter of healthy adults

The primary clinical symptom of Paralytic Shellfish Poisoning is acute paralytic illness produced by paralyzing toxins. Paralytic shellfish poison is formed by a mixture of phycotoxins and their toxicity is due to its reversible binding to a receptor site on the voltage-gated sodium channel on excitable cells, thus blocking neuronal transmission. We studied the effect of the gonyautoxin 2/3 epimers by local infiltration in the anal internal sphincter of healthy voluntary adults in order to reduce anal tone. The toxin was injected after prior clinical evaluation, anoscopy and anorectal manometry. Post injection clinical examination, electromyography and anorectal manometry were performed. Resting and voluntary contraction pressures were measured and the anorectal inhibitory and anocortical reflexes were tested by manometry. Blood and urine samples were obtained from each participant, and hemogram, basic metabolic panel, and urinalysis were done both before and one week after the injection. This study shows, for the first time, that gonyautoxin 2/3 reduces the anal tone by relaxing the anal sphincters in 100 % of the participants. Manometric recordings showed a significant decrease in anal maximal voluntary contraction pressure after the toxin injection, dropping to 55.2 ± 6.2 % and 47.0 ± 6.8 % (Mean Value ± Std.Dev.) of the baseline values at 2 minutes and at 24 hours respectively after the injection. Post-injection electromyography showed that activity of the muscle was abolished. We conclude that local administration of gonyautoxin 2/3 to the anal sphincter produces immediate relaxation and a statistically significant decrease in the anal tone (p <0.001)..

Isolation and purification of gonyautoxins from Alexandrium mimutum Halim / 药学学报

<p><b>AIM</b>To isolate and purify gonyautoxins from Alexandrium mimutum Halim Amtk2 strain.</p><p><b>METHODS</b>The ethanol extracts of culture Alexandriun minutum Halim Amtk2 were isolated by means of gel filtration chromatography, the toxin fraction obtained was then purified by ion exchange chromatography.</p><p><b>RESULTS</b>From 100 liter of cultivation liquid of Alexandrium mimutum Halim Amtk2 (6.74 +/- 0.31) x 10(9) cells were obtained. The ethanol extracts of Alexandriun minutum Halim purified by gel filtration chromatography obtained gonyautoxins mixture 29.59 mg. 4.06 mg of the mixture was further purified by two steps of ion exchange chromatography, and obtained pure GTX-4 (0.40 +/- 0.002) mg, GTX-1 (5.95 +/- 0.03) x 10(-2) mg, GTX-3 (6.92 +/- 0.05) x 10(-4) mg and GTX-2 (0.11 +/- 0.005) mg.</p><p><b>CONCLUSION</b>Pure gonyautoxins can be obtained by means of gel filtration chromatography and ion exchange chromatography from ethanol extracts of cultured Alexandriun minutum Halim Amtk2 strain.</p>

Effect of antibiotic treatment on toxin production by Alexandrium tamarense / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Impact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated.</p><p><b>METHODS</b>Pronounced changes in the activities of the algal culture were observed when the culture was treated with different doses of a mixture of penicillin and streptomycin.</p><p><b>RESULTS</b>In the presence of antibiotics at the initial concentration of 100 u/mL in culture medium, both algal growth and toxin yield increased markedly. When the concentration of antibiotics was increased to 500 u/mL, the microalgal growth was inhibited, but resumed in a few days to eventually reach the same level of growth and toxin production as at the lower dose of the antibiotics. When the antibiotics were present at a concentration of 1 000 u/mL, the algal growth was inhibited permanently.</p><p><b>CONCLUSIONS</b>The results indicate that antibiotics can enhance algal growth and toxin production not only through their inhibition of the growth and hence competition for nutrients, but also through their effects on the physiology of the algae.</p>

Paralytic Shellfish Poisoning by Saxitoxin: Two case reports

Paralytic shellfish poisoning results from consumption of mollusks that have fed on dinoflagellates capable of producing neurotoxins such as saxitoxin. The saxitoxin is concentrated in the shellfish and acts by decreasing sodium-channel permeability, thereby blocking neuronal transmission in skeletal muscles. Symptoms including paresthesia, perioral numbness, perioral tingling, nausea, vomiting, extremity numbness, extremity tingling, dizziness, ataxia, dysphagia, and weakness have been reported. In serious cases, respiratory hold may occur up to 6~24 hours after ingestion. Generally, the treatment for paralytic shellfish poisoning is supportive care, but mechanical ventilation is needed in serious cases acompanied by respiratory hold. We experienced two cases of paralytic shellfish poisoning. Respiratory hold was presented in one case and only mild paresthesia in the other case. After supportive management, including mechanical ventilation in former case, both patients were discharged without sequalae.

Intoxicación paralítica por moluscos: fenómeno de marea roja / Paralytic shellfish poisoning: red tide phenomenon

El fenómeno conocido como MAREA ROJA es causado por dinoflagelados del género GONYAULAX. Estos sintetizan toxinas que se aculmulan en moluscos bivalvos (mejillones y pepitonas) que al ser ingeridos por los humanos pueden causarle parálisis y la muerte.

Occurrence of tetrodotoxin and paralytic shellfish toxins in Phallusia nigra (Tunicata, Ascidiacea) from the brazilian coast

We have already shown the presence of guanidine neurotoxins in calcareous red algae and mussels collected in the Säo Sebastiäo channel State of Säo Paulo,Brazil). It is known that these neurotoxins comprise more than 25 analogues such as tetrodotoxin (TTX) and derivatives plus the paralytic shellfish toxins (PST) found in a variety of marine, freshwater and amphibious species. Filter feeding animals generally possess large amounts of these neurotoxins. The tunicates are sessil marine animals with a high rate of sea water filtration. The tunics and siphons of 50 specimens of Phallusia nigra were dissected and the visceral organs were immersed in methanol containing acetic acid 0.02 N ph 5.0. The extract was prepared by homogenization, filtration and the methanolic phase was concentrated under reduced pressure and defatted with chloroform. The polar phase was evaporated and the residue dissolved in deionized water for further purification in ionic-exchange resin column (Bio-Gel P-2) and HPLC analysis. The extract showed paralytic effects on mouse assay (26.9 MU/100mg) and on crustacean isolated nerve preparations. The chemical analysis for TTX and PST revealed toxins with retention times similar to gonyautoxins, saxitoxins and TTX. These findings are important to explain future toxin envenoming outbreaks on the Brazilian coast.