RESUMO

Causative species of Harmful Algal Bloom (HAB) and toxins in commercially exploited molluscan shellfish species are monitored weekly from four classified shellfish production areas in Perú (three in the north and one in the south). Okadaic acid (OA) and pectenotoxins (PTXs) were detected in hand-picked cells of Dinophysis (D. acuminata-complex and D. caudata) and in scallops (Argopecten purpuratus), the most important commercial bivalve species in Perú. LC-MS analyses revealed two different toxin profiles associated with species of the D. acuminata-complex: (a) one with OA (0.3⁻8.0 pg cell-1) and PTX2 (1.5⁻11.1 pg cell-1) and (b) another with only PTX2 which included populations with different toxin cell quota (9.3⁻9.6 pg cell-1 and 5.8⁻9.2 pg cell-1). Toxin results suggest the likely presence of two morphotypes of the D. acuminata-complex in the north, and only one of them in the south. Likewise, shellfish toxin analyses revealed the presence of PTX2 in all samples (10.3⁻34.8 µg kg-1), but OA (7.7⁻15.2 µg kg-1) only in the northern samples. Toxin levels were below the regulatory limits established for diarrhetic shellfish poisoning (DSP) and PTXs (160 µg OA kg-1) in Perú, in all samples analyzed. This is the first report confirming the presence of OA and PTX in Dinophysis cells and in shellfish from Peruvian coastal waters.

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RESUMO

El presente trabajo informa sobre características bioquímicas, actividad hemolítica, citotóxica y citolítica de tres fracciones del veneno de la anémona de mar Anthothoe chilensis. Los tentáculos de 78 ejemplares de A. chilensis, provenientes de la Isla Cabinza-San Lorenzo, Lima, fueron procesados obteniendo un filtrado, el cual se fraccionó por precipitación con tres puntos de saturación con acetona fría: I (20 por ciento), II (50 por ciento), III (80 por ciento). El filtrado mostró una concentración proteica de 1,8 mg/mL. Las pruebas de detección de carbohidratos totales demostraron la presencia de 1,401 mg de glucosa/ml de solución en el filtrado; mientras que la electroforesis en gel de poliacrilamida en presencia de dodecil sulfato de sodio (PAGE-SDS) evidenció proteínas de 14 a 94 kDa, de las cuales la mayor parte fueron glicoproteínas. Se encontró actividad hemolítica sobre eritrocitos humanos en las fracciones I y II. La fracción III tuvo la actividad fosfolipásica más alta. Las tres fracciones tuvieron una ligera actividad proteolítica sobre caseína siendo la más activa la fracción I. Los efectos citotóxico y citológico fueron evaluados aplicando el Ensayo de Toxicidad en Embriones de Erizo de mar (SET). Las anormalidades morfológicas fueron evaluadas a las 48 horas de desarrollo. Las anormalidades citológicas fueron evaluadas en el estadio de gástrula tardía. Las tres fracciones acetónicas produjeron daños citotóxicos y citológicos importantes en los embriones de erizo de mar. Los efectos sobre los embriones fueron retrasar su desarrollo y producir anomalías morfológicas como lisis de blástulas y exogastrulación. Los daños citológicos observados fueron núcleos heteropicnóticos, núcleos gigantes y espacios celulares anormales.

This paper reports on biochemical characteristics, hemolytic activity, cytolytic and citotóxic of three fractions venom from the sea anemone Anthothoe chilensis. The arms of 78 individuals of A. chilensis, from the Isle Cabinza-San Lorenzo, Lima, were treated to obtain a filtrate (Ft), which is divided by precipitation with three points of saturation with cold acetone: I (20 per cent), II (50 per cent), III (80 per cent). The Ft showed 1,8 mg/mL of protein concentration. Test of total carbohydrate detection showed the presence of 1,401 mg of glucose/ml of solution in Ft. SDS-PAGE showed proteins between 14 to 94 kDa, almost were glycoproteins. Hemolytic activity was assayed on human erythrocytes showing positive result the fractions II and I. The fraction III was highest phospholipasic activity. Evaluating the proteolytic activity on casein was observed that the fraction I has the highest activity. Cytotoxic and cytological effects were evaluated in sea urchin embryos Tetrapigus niger. The morphologic abnormalities were evaluated at 48 hours of developing. Cytological abnormalities were observed at late gastrule stage. All acetonic fractions caused abnormalities in embryonic development. Delay of development, morphologic abnormalities like lysis at the blastula and exogastrulation, cytological abnormalities like heteropycnosis, giant nuclei and cellular spaces were observed.

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