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New targets in diarrhetic shellfish poisoning control.

The toxic profile of dinoflagellates varies even among identical species, raising an interesting question about the mechanism of toxin regulation and production. In consequence, it also poses a considerable problem in their control methods. In this paper, we report on the isolation and structural elucidation of several new ester derivatives of okadaic acid (OA) from artificial cultures of the genus Prorocentrum. These new compounds enlarge the range of target molecules that must be considered in the monitoring programs.

Animals↗

Detection and identification of spirolides in norwegian shellfish and plankton.

Mussels sampled in the spring of 2002 and 2003 from Skjer, a location in Sognefjord, Norway, tested positive in the mouse bioassay for lipophilic toxins. The symptoms, which included cramps, jumping, and short survival times (as low as 4 min), were not characteristic of toxins previously observed in Norway. A survey of the algae present at the aquaculture sites showed that the toxicity correlated with blooms of Alexandrium ostenfeldii. Up to 2200 cells/L were found at the peak of one bloom. In Canadian waters, this alga is known to be a producer of the cyclic imine toxins, spirolides. Analysis of mussel extracts from Skjer in the spring of 2002 and 2003, using liquid chromatography tandem mass spectrometry, revealed the presence of several new spirolides. The same compounds were also found in algal samples dominated by A. ostenfeldii, which had been sampled from Skjer in February 2003. A large-scale extraction of mussel digestive glands and chromatographic fractionation of the extracts allowed the isolation and structure elucidation of the main spirolide, 20-methyl spirolide G, with a molecular weight of 705.5. This is the first confirmed occurrence of spirolides in mussels and plankton from Norway.

Acetals↗

Low incidence of Vibrio vulnificus among Vibrio isolates from sea water and shellfish of the western Mediterranean coast.

A specific search for Vibrio vulnificus in natural marine samples from the Spanish Mediterranean Sea was carried out by nested PCR and cultural approaches using thiosulphate-citrate-bile salts-sucrose agar (TCBS) and cellobiose-polymixin B-colistin agar (CPC), incubated at 40 degrees C, as selective media. Presumptive colonies were identified by PCR using specific primers against 23S rRNA sequences. This species was isolated from sea water and edible bivalves, mainly after preenrichment in alkaline peptone water (APW) at 40 degrees C followed by CPC agar. None of the V. vulnificus isolates identified corresponded to serovar E. Dominant Vibrio species on directly inoculated TCBS plates incubated at 25 degrees C were V. splendidus below 20 degrees C and V. harveyi and V. mediterranei above that temperature. Low percentages of several pathogenic vibrios were recorded but V. vulnificus was never recovered at this incubation temperature. The incidence of this species in the samples studied was lower than that described for other geographical areas, probably due to the high salinity values of the Mediterranean Sea.

Animals↗

Structural studies on glycolipid of shellfish. III. Novel glycolipids from Turbo cornutus.

Five kinds of sphingoglycolipids were isolated from Turbo cornutus. Four of them were a series of novel glycolipids consisting only of galactose. The structures of these glycolipids were studied by methylation analysis, periodate oxidation, enzymatic degradation, and proton magnetic resonance spectroscopy. Three glycolipids were characterized as galactosyl(beta 1 leads to 1)ceramide, galactosyl(beta 1 leads to 6)galactosyl(beta 1 leads to 1)ceramide, and galactosyl(beta 1 leads to 6)galactosyl(beta 1 leads to 6)galactosyl(beta 1 leads to 1)ceramide. Data indicating that the 4th glycolipid might be the tetragalactosyl derivative of this series were obtained. The carbohydrate moiety of the 5th glycolipid, in contrast, was composed of fucose, galactose, glucose and N-acetylglycosamine in a molar ratio of 1 : 2 : 1 : 1.

Carbohydrates↗

Release of tetrodotoxin and paralytic shellfish poison from puffer liver by RNase.

A nontoxic high-molecular fraction was separated from the extracts of toxic liver of a puffer, Takifugu poecilonotus, by Sephadex G-50 gel filtration. This fraction became toxic when digested with RNase T2. The toxins were partially purified by activated charcoal treatment, followed by chromatography on Bio-Gel P-2 and Bio-Rex 70, and were analyzed by TLC and electrophoresis. The results showed that most of the toxicity is accounted for by tetrodotoxin, and the remainder by saxitoxin and other unidentified toxins. The corresponding high-molecular fraction separated from nontoxic liver of another puffer, T. rubripes, did not release any toxin on RNase digestion.

Animals↗