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Results for “Shellfish Poisoning”

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PubMed1996

Development of a manganese dioxide solid-phase reactor for oxidation of toxins associated with paralytic shellfish poisoning.

Using manganese dioxide as a solid-phase oxidant, a post-column HPLC reactor for paralytic shellfish poison toxins was constructed and evaluated. Operating parameters such as reaction temperature and pH, flow-rate, reactor column size, and MnO2 particle size were studied. Based on a 3:1 signal-to-noise ratio the limits of detection for the non-hydroxylated toxins were in the sub-nanogram range, while the hydroxylated toxins were ten times less detectable. The limit of detection for saxitoxin was about 0.1 ng per injection. The repeatability of replicate injections was < or = 10% (relative standard deviation). The system was used to analyze extracts of shellfish and plankton yielding results that were in agreement with those obtained by established analytical methods. Extracts of shellfish contaminated at the regulatory limit of 0.8 microgram/g were successfully analyzed.

Animals↗

PubMed1990

[Paralytic shellfish poisoning by Spondylus calcifer contaminated with Pyrodinium bahamense, Costa Rica, 1989-1990].

This paper describes an outbreak of paralytic shellfish poisoning (PSP), affecting human populations on the Pacific Coast of Costa Rica in October 1989. Numbness in arms, face and legs occurred 30 to 45 minutes after ingestion of the large clam Spondylus calcifer. Paralysis of legs and respiratory symptoms followed, often persisting for one week. Large amounts of the dinoflagellate Pyrodinium bahamense were found in the intestine of the mollusk. A toxin was detected in crude or filtered and heated macerates of intestine, muscle, mantle and hepatopancreas of S. calcifer, and to a lesser extent Tagelus sp., by injection of its crude or diluted extracts in white mice. The effects in mice consisted in paralysis and asphyxia generally leading to death in less than 5 minutes, compatible with saxitoxin. Mice were killed by the toxin in macerates diluted 1:100 to 1:1000. No toxin was detected in Anadara tuberculosa (Bivalvia) or in peneids. Prevention rests on intersectoral actions between state and private sectors in charge of fishing, distribution and marketing of shellfish, as well as on education of the population at large.

Adult↗

PubMed2003

Improved solid-phase extraction procedure in the analysis of paralytic shellfish poisoning toxins by liquid chromatography with fluorescence detection.

The analysis of shellfish extracts for the determination of paralytic shellfish poisoning (PSP) toxins by liquid chromatography with fluorescence detection repeatedly showed the presence of a compound suspected to interfere with gonyautoxin 4. The first aim of this study was to confirm by liquid chromatography coupled to tandem mass spectrometry that this compound was not gonyautoxin 4. The second part of this work was to improve a nonvolumetric C(18) solid-phase extraction (SPE) procedure to evaluate the removal of the interference associated with the recovery of PSP toxins. The cleanup procedure was modified into a volumetric SPE procedure and proved to efficiently and totally remove the interference while recovering from 78 to 85% of the PSP toxins available as commercial standards (saxitoxin, neosaxitoxin, gonyautoxins 1-4) and considered as major PSP toxins in human intoxication, with 85% recovery for gonyautoxin 4. The efficiency of this cleanup procedure was checked on shellfish extracts containing this interference and originating from France and Turkey.

Chromatography, Liquid↗

PubMed2002

kappa-Carrageenan gel as agent to sequester paralytic shellfish poison.

The action of k-carrageenan gel to sequester paralytic shellfish poison (PSP) was tested and characterized. When an extract from a Philippine strain of Pyrodinium bahamense var. compressum was used as PSP solution, the PSP-sequestering property of kappa-carrageenan gel was found to be dependent on gel surface area, interaction time, and polysaccharide concentration. The interaction was also found to be affected by high concentrations of monovalent cations. The characteristics of kappa-carrageenan as a PSP-sequestering agent all point to cation exchange as its mechanism of action. It is also proposed that the polysaccharide gel can be utilized as an agent to alleviate PSP intoxication.

Journal Article↗

PubMed1993

[Clinical analysis of paralytic shellfish poisoning following ingestion of oysters].

We describe five patients of paralytic shellfish poisoning (PSP) following ingestion of oysters in January 1991, in Nagato, Yamaguchi. The five patients (four men and one woman, age range:37-80 years) developed symptoms three-nine hours after consuming oysters. Symptoms included paresthesia of the mouth and tongue (three), paresthesia of the extremities (five), and quadriparesis (five). The median duration of neurological symptoms was 16 hours. All five patients completely recovered. Although PSP is rare as compared with globe fish poisoning in Japan, the fatality rate of PSP is 8%-9%, with deaths occurring in one-12 hours secondary to respiratory failure. PSP is an important disorder for differential diagnosis of acute paralytic illnesses, such as globe fish poisoning, botulism, acute polyradiculoneuropathy, fisher syndrome, myasthenia gravis, and periodic paralysis.

Adult↗

PubMedDate not supplied

Assessment of MIST Alert, a commercial qualitative assay for detection of paralytic shellfish poisoning toxins in bivalve molluscs.

A recently developed commercial rapid test kit (MIST Alert) was assessed for determination of the presence of paralytic shellfish poisoning (PSP) toxins in shellfish. Several commercially important shellfish species obtained from the UK shellfish toxin monitoring program, containing a range of total PSP toxicities as determined by the mouse bioassay (MBA), were tested. The kit detected toxin in all samples containing the European Community tolerance level of 80 microg saxitoxin (STX) equivalents/100 g shellfish flesh as determined by the MBA. With one exception, the kit detected toxin in all samples that contained >40 microg STX equivalents/100 g according to the MBA. Among samples in which the MBA did not detect toxin, the kit disagreed in 25% of the tests, although further analysis by liquid chromatography (LC) and MBA of some samples confirmed the presence of toxins. These results suggest that MIST Alert may be suitable as an initial screen for PSP toxins as part of routine monitoring programs, thereby greatly reducing the number of MBAs. Trials were also performed by nonscientific personnel to evaluate the ease of use and interpretation of results obtained by MIST Alert. The results indicated that the kits could be readily used and accurately interpreted by individuals with no technical or scientific background.

Animals↗

PubMed2001

Determination of paralytic shellfish poisoning toxins by high-performance ion-exchange chromatography.

An efficient LC method has been developed for the determination of paralytic shellfish poisoning (PSP) toxins based on ion-exchange chromatographic separation of the toxins followed by electrochemical post-column oxidation and fluorescence detection as well as mass spectrometric (MS) detection. The method can be applied to the determination of PSP toxins in phytoplankton and to control seafood for PSP content.

Chromatography, High Pressure Liquid↗

PubMedDate not supplied

Immunoassay methods for paralytic shellfish poisoning toxins.

The current status of immunochemical techniques for analysis of paralytic shellfish poisoning (PSP) toxins is summarized. Important aspects regarding production of the biological reagents necessary for immunochemical methods, the characteristics of polyclonal and monoclonal antibodies against saxitoxin and neosaxitoxin, and the importance of test sensitivity and specificity are discussed. Applications of immunochemical techniques for PSP toxins include microtiter plate enzyme immunoasays and enzyme-linked immunofiltration assays for toxin detection, and immunoaffinity chromatography (IAC) for sample extract cleanup. A major advantage of enzyme immunoassay (EIA) is simplicity and rapidity of the test procedure, and higher sensitivity than other methods. However, quantitative agreement between EIA and mouse bioassay is dependent on antibody specificity and the toxin profile in the shellfish; thus, both over- and underestimation of total toxicity may occur. For screening purposes, however, EIAs offer major advantages over the mouse bioassay, which is criticized in Europe because of animal welfare. A major application of antibodies against PSP toxins is their use for extract cleanup by IAC, which gives highly purified extracts, thereby enhancing determination of PSP toxins by conventional physicochemical methods such as liquid chromatography. IAC can also be used to isolate PSP toxins for preparation of analytical standard solutions.

Animals↗

PubMed1997

Optimization of sample cleanup procedure for determination of diarrhoeic shellfish poisoning toxins by use of experimental design.

In routine monitoring of diarrhoeic shellfish poison (dinophysistoxin-1 and okadaic acid) there appeared to be an inconsistency between the mouse bioassay and existing chemical analysis based on liquid chromatography. The sample cleanup procedure has been subject to minor modifications in an effort to overcome the problem. However, further studies have appeared necessary and in this study all experimental factors that can influence the sample cleanup using solid-phase extraction columns prior to the LC analysis have been evaluated by use of experimental statistical design in order to understand the effect of the various factors and to optimize the conditions for recovery of the toxins. Based on our experiments we suggest using a solid-phase extraction silica column of 100 mg in the sample cleanup procedure and washing solvents composed of dichloromethane instead of chloroform to minimize the effect of stabilizing alcohol. It is sufficient to apply 7.5 ml hexane-dichloromethane (1:1) to the column in the first washing step and 2.5 ml of dichloromethane in the final washing. Elution is complete by use of 2.5 ml chloroform with 3.5% methanol. Toxic shellfish tested by this procedure confirmed the mouse bioassay.

Animals↗

PubMed2005

Isodomoic acid C, an unusual amnesic shellfish poisoning toxin from Pseudo-nitzschia australis.

An unusual isomer of domoic acid (1), isodomoic acid C (2), has been found in New Zealand shellfish contaminated by amnesic shellfish poisoning (ASP) toxins and was shown to be produced by a local strain of the pennate diatom Pseudo-nitzschia australis. A bulk culture of this strain was used to isolate 2. The structure was determined from spectroscopic data and was shown to correspond to that of 2 from a Japanese red seaweed, the only other reported occurrence of this compound. The affinity of 2 for GluR6 glutamate receptors was 240-fold lower than for 1, indicating low neurotoxic potential.

Amnesia↗

PubMed1999

Brevetoxin B4 isolated from greenshell mussels Perna canaliculus, the major toxin involved in neurotoxic shellfish poisoning in New Zealand.

A new brevetoxin B analog, brevetoxin B4 (BTXB4), was isolated as the major toxin in greenshell mussels, Perna canaliculus, collected in New Zealand at the time of the neurotoxic shellfish poisoning incident. The new analog accounted for nearly two-thirds of the mouse lethality of the shellfish and was determined to be a mixture of N-myristoyl-BTXB2 and N-palmitoyl-BTXB2.

Animals↗

PubMed1998

A high-throughput, microtiter plate assay for paralytic shellfish poisons using the saxitoxin-specific receptor, saxiphilin.

An isoform of the paralytic shellfish poison (PSP)-specific receptor saxiphilin, from the tropical centipede Ethmostigmus rubripes, was used as the basis for a radiometric, high-throughput, microtiter plate assay for this group of toxins. Characterization of the assay revealed that it was able to detect several representatives from the various structural PSP subgroups and yet was insensitive toward tetrodotoxin. To test the utility of the assay as a seafood toxin-monitoring tool, the assay was subjected to a variety of marine organism extracts, some of which were known to contain PSPs, and whole extract toxicity expressed as STX equivalents (STXeq) was measured by two methods: First, by comparison of values from a screening assay with a standard STX inhibition curve and, second, for highly active extracts, by calculation using the IC50 from a full inhibition curve of the extract. For extracts which could be quantified by both methods, there was almost 100% correlation between the derived values. STXeq derived by both methods from the bioassay highly correlated with absolute toxin quantities from HPLC analysis.

Amphibian Proteins↗

PubMed2002

Matrix effect and correction by standard addition in quantitative liquid chromatographic-mass spectrometric analysis of diarrhetic shellfish poisoning toxins.

An evaluation of the feasibility of liquid chromatography-mass spectrometry (LC-MS) with atmospheric pressure ionization was made for quantitation of four diarrhetic shellfish poisoning toxins, okadaic acid, dinophysistoxin-1, pectenotoxin-6 and yessotoxin in scallops. When LC-MS was applied to the analysis of scallop extracts, large signal suppressions were observed due to coeluting substances from the column. To compensate for these matrix signal suppressions, the standard addition method was applied. First, the sample was analyzed and then the sample involving the addition of calibration standards is analyzed. Although this method requires two LC-MS runs per analysis, effective correction of quantitative errors was found.

Animals↗

PubMedDate not supplied

Liquid chromatographic determination of domoic acid in mussels, using AOAC paralytic shellfish poison extraction procedure: collaborative study.

A liquid chromatographic method using the AOAC paralytic shellfish poison (PSP) extraction procedure for domoic acid, a marine toxin, in mussel tissue was collaboratively studied in 10 laboratories. Domoic acid is extracted by boiling the homogenized tissue for 5 min with 0.1N HCl. The mixture is cooled, diluted to a known volume, and then centrifuged. An aliquot of the supernate is diluted, filtered, and analyzed by reverse-phase liquid chromatography with a mobile phase containing acetonitrile and water adjusted to about pH 2.5. Each collaborator received a prepared standard solution, a practice sample, and 7 randomly numbered unknown samples (1 blank mussel tissue, 1 spiked at 14.1 micrograms domoic acid/g, 1 spiked at 18.9 micrograms/g, and duplicate samples with naturally incurred domoic acid at 75 micrograms/g and at 186 micrograms/g). Five of the laboratories had little or no experience in domoic acid analysis. Ten of 11 laboratories completed the study and submitted results. Two individual values out of a total of 70 were found to be outliers. Mean recovery of domoic acid from the spiked extracts was 75%. Relative standard deviations between laboratories (RSDR) ranged from 7.5 to 19.4%; within-laboratory RSDs (RSDr) for the 2 blind duplicate pairs were 1.9 and 4.8%. The detection limit was about 1 microgram domoic acid/g. The method has been adopted official first action by AOAC.

Animals↗

PubMedDate not supplied

Evaluation of a postcolumn electrochemical reactor for oxidation of paralytic shellfish poison toxins.

A liquid chromatographic method using a postcolumn electrochemical reactor that oxidizes paralytic shellfish poison toxins to fluorescent derivatives has been developed. Several experimental parameters, including pH and oxidation potential, were investigated. For nonhydroxylated toxins, the sensitivity improved with increasing pH and voltage. At optimum operating conditions, the sensitivity for saxitoxin and gonyautoxins 2 and 3 was an order of magnitude greater than that for neosaxitoxin and B1 and 2 orders of magnitude greater than that for B2. The limit of detection for saxitoxin was 0.10 ng (signal-to-noise ratio, 3:1). Electrochemical oxidation products were similar to those formed in the prechromatographic periodate oxidation method. Shellfish and plankton extracts were analyzed with the electrochemical system, and results agreed well with those obtained with established methods. Shellfish samples contaminated at the regulatory limit of 0.8 microgram/g were readily analyzed by the method.

Animals↗

PubMed2004

Paralytic shellfish poisoning toxins in green mussels (Perna viridis) from the Gulf of Paria, Trinidad.

Paralytic shellfish poisoning (PSP) toxins were determined in green mussels (Perna viridis) collected from one collection site in the Gulf of Paria in Trinidad in 1999 and 2000. Aqueous extracts of PSP were purified by passage through C-18 SPE cartridges, oxidized with peroxide and periodate, respectively, then analyzed by HPLC with fluorescence detection. This procedure provided rapid and highly sensitive screening of samples for PSP toxins. Further purification of PSP-containing extracts using COOH SPE cartridges resulted in the separation and identification of individual PSP toxins. The method of analysis was validated by spike and recovery experiments, with 85-103% recoveries of mixed toxins. PSP toxins determined in our samples in both years were GTX2,3, dcGTX2,3, STX, and dcSTX, while GTX1,4 and NeoSTX were only identified in 1999 and 2000, respectively. In 1999, GTX1,4, GTX2,3 and dcGTX2,3 predominated, as compared to NeoSTX, GTX2,3 and dcGTX2,3 in 2000. However, mussel samples in 2000 contained higher total concentrations of detected PSP toxins than those of 1999. These results represent the first identification of specific PSP toxins in local shellfish and provide a basis for effective monitoring and control of these toxins in Trinidad.

Animals↗

PubMed1999

A receptor binding assay for paralytic shellfish poisoning toxins: recent advances and applications.

We recently described a high throughput receptor binding assay for paralytic shellfish poisoning (PSP) toxins, the use of the assay for detecting toxic activity in shellfish and algal extracts, and the validation of 11-[3H]-tetrodotoxin as an alternative radioligand to the [3H]-saxitoxin conventionally employed in the assay. Here, we report a dramatic increase in assay efficiency through application of microplate scintillation technology, resulting in an assay turn around time of 4 h. Efforts are now focused on demonstrating the range of applications for which this receptor assay can provide data comparable to the more time consuming, technically demanding HPLC analysis of PSP toxins, currently the method of choice for researchers. To date, we have compared the results of both methods for a variety of sample types, including different genera of PSP toxin producing dinoflagellates (e.g. Alexandrium lusitanicum, r2 = 0.9834, n = 12), size-fractioned field samples of Alexandrium spp. (20-64 microm; r2 = 0.9997, n = 10) as well as its associated zooplankton grazer community (200-500 microm: r2 = 0.6169, n = 10; >500 microm: r2 = 0.5063, n = 10), and contaminated human fluids (r2 = 0.9661, n = 7) from a PSP outbreak. Receptor-based STX equivalent values for all but the zooplankton samples were highly correlated and exhibited close quantitative agreement with those produced by HPLC. While the PSP receptor binding assay does not provide information on toxin composition obtainable by HPLC, it does represent a robust and reliable means of rapidly assessing PSP-like toxicity in laboratory and field samples. Moreover, this assay should be effective as a screening tool for use by public health officials in responding to suspected cases of PSP intoxication.

Animals↗

PubMed1993

Characterization of the oxidation products of paralytic shellfish poisoning toxins by liquid chromatography/mass spectrometry.

Periodate oxidation of the toxins responsible for paralytic shellfish poisoning (PSP) yields fluorescent products that are useful for trace analysis by reversed-phase liquid chromatography. Some toxins are oxidized to the same product, while others are transformed to a mixture of products; in all cases, the products are too unstable to be isolated preparatively. A mobile phase containing heptafluorobutyric acid as an ion-pair agent was developed to improve compatibility with liquid chromatography/mass spectrometry (LC/MS) analysis. Several LC/MS techniques were investigated: chemical ionization with the moving belt interface, atmospheric-pressure chemical ionization, thermospray, continuous-flow fast-atom bombardment, and ion-spray. Only LC/ion-spray MS was able to provide spectra of the oxidized products. The products were characterized as purines resulting from: (a) breakage of the C4-C12 bond and aromatization of the remaining rings (loss of 4 hydrogens); (b) partial elimination of a carbamoyl or N-sulfocarbamoyl side-chain (this is promoted by the presence of an N-1-hydroxy group); and (c) partial hydrolysis of an N-sulfocarbamoyl side-chain to a carbamoyl group.

Gas Chromatography-Mass Spectrometry↗