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Levels of male-specific RNA bacteriophage and Escherichia coli in molluscan bivalve shellfish from commercial harvesting areas.

AIMS: Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator organism, male-specific RNA (FRNA) bacteriophage has been proposed for this role. This study compared the distribution of E. coli and FRNA bacteriophage in shellfish harvesting areas. METHODS AND RESULTS: A total of 608 shellfish samples from 49 shellfish harvesting areas were analysed for E. coli and FRNA bacteriophage using standard published methods. The geometric mean concentration of FRNA bacteriophage in all samples was over three times greater than that of E. coli (1800 and 538 counts/100 g for FRNA bacteriophage and E. coli, respectively). In contrast to E. coli, FRNA bacteriophage concentrations were strongly influenced by season with a geometric mean count of 4503 PFU/100 g in the winter (October-March) compared with 910 PFU/100 g in the summer (April-September). CONCLUSIONS: FRNA bacteriophage were present in shellfish at higher concentrations than E. coli. Elevated levels of FRNA bacteriophage observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in the UK. Levels of FRNA bacteriophage found in many shellfish from category B harvesting areas would not be eliminated by conventional treatment processes. SIGNIFICANCE AND IMPACT OF THE STUDY: Data from this study will inform future proposals to introduce FRNA bacteriophage as an indicator of the viral risk associated with shellfish.

Animals↗

Derivation of shellfish harvest reopening criteria following the New Carissa oil spill in Coos Bay, Oregon.

Oil spills in Alaska, California, Maine, and other states have raised concerns regarding potential contamination of fish and shellfish, and have led to temporary closures of seafood harvests while health risks are assessed. Lacking standardized protocols, these assessments are generally ad hoc, site-specific efforts, with significant differences in risk evaluation criteria. This article describes the response of a state health agency to shellfish contamination following an oil spill on the Oregon coast, and discusses some of the factors that can complicate the evaluation of potential health risks from consumption of oil-contaminated shellfish. On 4 February 1999, the Japanese-owned cargo ship M/V New Carissa, carrying an estimated 400,000 gallons of light diesel and heavy fuel oil, ran aground 2 miles north of Coos Bay, Oregon. Damage to the ship's hull from the grounding and pounding surf caused the release of an estimated 25,000 to 70,000 gallons of oil. Concern for potential contamination of local recreational shellfish and commercial oyster beds prompted the Oregon Department of Agriculture (ODA) to close shellfish harvesting in Coos and Douglas counties. ODA requested assistance from the Oregon Health Division in the derivation of risk-based criteria for reopening the shellfish harvest. Criteria were developed for the primary contaminants of concern, polycyclic aromatic hydrocarbons (PAHs) expressed as total benzo[a]-pyrene (BaP) equivalents. "Safe" (<10 microg/kg) and "unsafe" (>45 microg/kg) BaP equivalent levels were derived based on upper end (32.5 g/d) and average (7.5 g/d) estimates of shellfish consumption, respectively. Composite samples of oysters, clams, and mussels (15-20 per composite) were collected from target areas and analyzed for PAHs by gas chromatography/mass spectroscopy (GC/MS). Carcinogenic PAHs were converted to total BaP equivalents (wet weight) and compared with criteria. Two oyster samples, collected from a slough off of Coos Bay, contained 33.9 and 34.5 microg/kg BaP equivalents; all other samples had less than 10 microg/kg BaP equivalents. An evaluation of the PAH profiles in the two higher oyster samples indicated a primary source other than the New Carissa oil. Because shellfish sample BaP equivalents attributable to the New Carissa oil spill did not exceed 10 microg/kg, shellfish harvesting was reopened on 4 March 1999. This study revealed some of the inherent difficulties in attempting to quantify health risks from contaminated shellfish following an oil spill and demonstrated the clear need for standardized protocols for responding to such events.

Animals↗

A nested reverse transcriptase PCR assay for detection of small round-structured viruses in environmentally contaminated molluscan shellfish.

We describe the evaluation of a nested reverse transcriptase PCR (RT-PCR) procedure for the detection of small round-structured viruses (SRSVs) in molluscan shellfish and the application of this assay for the detection of SRSVs in commercially produced shellfish and in shellfish implicated in outbreaks of gastroenteritis. The range of virus strains detected and the sensitivity of detection were evaluated by using a representative panel of 21 well-characterized SRSV strains. The nested RT-PCR detected 15 of 21 SRSVs, demonstrating that the assay detects a broad range of SRSVs including strains from both genogroup I and genogroup II. Seeding experiments showed the nested RT-PCR assay to be 10 to 1,000 times more sensitive than the single-round RT-PCR assay for the detection of SRSV in shellfish. SRSV-contaminated samples were identified by nested RT-PCR for shellfish grown in polluted harvesting areas and for shellfish associated with outbreaks of gastroenteritis which were negative by a previously described single-round RT-PCR. The assay was shown to be effective for investigation of virus elimination during commercial shellfish processing procedures such as depuration and relaying and has potential applications for monitoring at-risk shellfish harvesting areas, for investigation of SRSV contamination in shellfish from producers linked to gastroenteritis outbreaks, and for the direct detection of virus in shellfish implicated in outbreaks.

Polymerase Chain Reaction↗

[Carrying capacity of shellfish culture in Dadeng Island sea area of Xiamen].

To fully and rationally exploit local living marine resources while have a sustainable, efficient and healthy development of shellfish culture in the Dadeng Island sea area of Xiamen, this paper determined and analyzed the related model parameters of this area, including chlorophyll a, primary productivity, phytoplankton organic carbon tent, wild filter feeder yields in subtidal and intertidal zones and suspension culture area, cultured shellfish filtration rate and organic carbon content, shellfish's total weight to fresh meat ratio, and adopted the Nutrient namic Model and Coastal Waters' Energy Flow Analysis Model to estimate the ecological capacity of shellfish this area, from which, the wild filter feeder yields were deducted for estimating shellfish carrying capacity. model established by Fang Jianguang was also used to estimate the shellfish carrying capacity. Statistics analysis was used to estimate the suitable culture area of shellfish and other species, aiming at limiting local shellfish ture and optimizing the culture of various species mollusks. According to the estimation of the three models, shellfish carrying capacity in this area should be 35,248-39,990 tons, with an average of 37,488 tons, 140,008 x 10(4) - 158,850 x 10(4) individuals, averaging 148,903 x 10(4). The theoretically suitable culture area 2 145 hm2, 1,900 hm2 for Ostreidae, 81 hm2 for razor clam (Sinonovacula constricta), 20 hm2 for blood (Tegillarca granosa), and 144 hm2 for musculus (Musculus senhousei). In 2000, the actual culture area of shellfish and other species in the waters around Dadeng surpassed the estimated suitable culture area. It is proposed that some measures should be taken to reduce the overexploited area.

Animals↗

Paralytic shellfish poisoning in southern China.

The rapidly expanding mariculture and commercial region along the southern coast of China has experienced sporadic outbreaks of paralytic shellfish poisoning for nearly 30 years, yet virtually nothing is known of the nature of that toxicity or of the causative organisms. This study presents the first direct comparisons of the high performance liquid chromatography toxin composition profiles of shellfish implicated in paralytic shellfish poisoning outbreaks in Daya Bay with Alexandrium tamarense cultures established from those waters. The three cultures that were analyzed produced an unusually high proportion of the low potency N-sulfocarbamoyl toxins C1 and C2 (nearly 90% of the total), and only trace quantities of the other saxitoxin derivatives. Total toxicity was thus very low with mild acid extraction, ranging between 7.2 and 12.7 fmole cell-1, or 0.7-0.9 pg saxitoxin equiv. cell-1. Following acid hydrolysis using the standard AOAC extraction method, the dominant toxins in the cultures were gonyautoxins 2 and 3 and decarbamoyl gonyautoxins 2 and 3. Total potency increased fourfold to 2.6-3.4 pg saxitoxin equiv. cell-1 following acid hydrolysis. These cultures are thus at the low end of the range of toxicities recorded for members of the A. tamarense species complex. Two scallop samples and one mussel sample collected from Daya Bay during paralytic shellfish poisoning episodes in 1990 and 1991 were also analyzed following the AOAC extraction procedure. The toxin profiles were similar for the three shellfish samples, in that the same suite of toxins were present in each, but the relative proportion of those toxins varied. The dominant toxins were gonyautoxins 2 and 3 and toxins C1-C4. Total toxicity was 336 and 654 micrograms saxitoxin per 100 g meat for the scallop samples, and 723 for the mussels. Toxins C3,4 were present in the shellfish at up to 22 mole%, but were not detected in cultures, even when mild acid was used for extraction. Despite the otherwise similar nature of the culture versus the shellfish toxin signatures, the presence of C3,4 indicates that another strain or species of Alexandrium, or possibly a paralytic shellfish poisoning-producing species of another genus was responsible for the 1990 and 1991 paralytic shellfish poisoning outbreaks in Daya Bay. Since the cultures analyzed were of low intrinsic toxicity, A. tamarense may be more widespread along the south coast of China than is suggested by the sporadic pattern of past paralytic shellfish poisoning outbreaks. Blooms with high cell density are required to generate sufficient toxin to be dangerous. The alarming increase in algal blooms in Chinese waters due to persistent and growing pollution may make these low toxicity populations more problematic in the future.

Animals↗

Contamination of Atlantic coast commercial shellfish with Cryptosporidium.

Shellfish (oysters and/or clams) were obtained from 37 commercial harvesting sites in 13 Atlantic coast states from Maine to Florida and one site in New Brunswick, Canada. Gill washings from each of 25 shellfish at each site were examined by immunofluorescence microscopy (IFA) for oocysts of Cryptosporidium. Gill washings from another 25 shellfish at each site were grouped into five pools of five shellfish each. DNA from each pool was utilized for PCR and genotyping. Oocysts were found in 3.7% of 925 oysters and clams examined by IFA in shellfish from New Brunswick and 11 of 13 states. Cryptosporidium DNA was detected by PCR in 35.2% of 185 pools. Cryptosporidium parvum genotypes 1 and 2, and Cryptosporidium meleagridis,all of which have been identified in infected humans, were identified at 37.8% of the sites. Gill washings from every site were tested for the presence of infectious oocysts by biological assay in neonatal BALB/c mice but no mice were found infected, suggesting that either the oocysts were no longer infectious or infections in mice were below the level of detection. Collectively, these findings indicate that Cryptosporidium species, indicative of pollution from human and animal feces and potentially infectious for humans, were found in commercial shellfish from 64.9% of sites examined along the Atlantic coast by either microscopy or molecular testing. Previous reports link periods of high rainfall with the elevated numbers of pathogen contaminated shellfish. Because shellfish in the present study were examined during a period of exceptionally low precipitation, the data are thought to underestimate the number of Cryptosporidium contaminated shellfish likely to be found during periods of normal or above normal precipitation.

Animals↗

Paralytic shellfish poisoning in Alaska: a 20-year retrospective analysis.

Outbreaks of paralytic shellfish poisoning have occurred worldwide. The authors reviewed records at the Alaska Division of Public Health to determine the epidemiologic characteristics of this disease. To assess risk factors for illness, the authors conducted a case-control study. A case was defined as illness compatible with paralytic shellfish poisoning within 12 hours of the consumption of shellfish, and a control was defined as a non-ill participant at a meal in which at least one case occurred. The authors documented 54 outbreaks of paralytic shellfish poisoning involving 117 ill persons from 1973 to 1992. One person died, four (3%) required intubation, and 29 (25%) required an emergency flight to a hospital. Outbreaks occurred with multiple shellfish species, during all four seasons, and at many locations. During the case-control study, illness was not associated with the shellfish toxin level, method of preparation, dose, race, sex, or age; alcohol consumption was associated with a reduced risk of illness (odds ratio = 0.05; p = 0.03). Although paralytic shellfish poisoning causes significant illness, the authors could not identify risk factors with clear implications for prevention strategies. This suggests that shellfish from uncertified beaches should not be eaten. Alcohol may protect against the adverse effects of paralytic shellfish poison.

Adolescent↗

Bioavailability of cadmium from shellfish and mixed diet in women.

Dietary intake and uptake of cadmium (Cd) were studied in nonsmoking women, 20-50 years of age, consuming a mixed diet low in shellfish (N = 34) or with shellfish once a week or more (N = 17). Duplicate diets were collected during 4 consecutive days for the determination of Cd content. The women kept detailed dietary records, and the intake of energy and various nutrients was calculated. The shellfish diets (median 22.3 micrograms Cd/day) contained twice as much Cd as the mixed diets (median 10.5 micrograms Cd/day; p < 0.0001). Cadmium in feces corresponded to 100 and 99% of that in duplicates of shellfish diets and mixed diets, respectively, indicating a low average absorption of the dietary Cd. In spite of the differences in the daily intake of Cd, there was no statistically significant difference in the concentrations of Cd in blood (B-Cd, shellfish group 0.25 micrograms/liter, mixed diet group 0.23 micrograms/liter) or urine (U-Cd, 0.10 micrograms Cd/liter in both groups). This indicates a lower absorption of Cd in the shellfish group than in the mixed diet group or a difference in the kinetics. A higher gastrointestinal absorption of Cd in the mixed diet group could partly be explained by lower body iron stores as measured by the concentrations of serum ferritin (S-fer, median 18 micrograms/liter, compared to 31 micrograms/liter in the shellfish group). In the mixed diet group, S-fer was negatively correlated with B-Cd and the main determining for B-Cd besides U-Cd in the multiple regression analysis, indicating an increased absorption of Cd at low body iron stores. When women with S-fer exceeding 20 micrograms/liter were compared, the higher dietary intake of Cd in the shellfish group compared to the mixed diet group (24 versus 10 micrograms/day) resulted in higher B-Cd (0.26 versus 0.16 micrograms/liter), although not in proportion to the difference in Cd intake. Thus, there seems to be differences in the bioavailability and/or kinetics of dietary Cd related to the type of diet. This is, to our knowledge, the first study where the influence of various types of diets and nutritional factors on the intake and uptake of cadmium in human subjects has been studied.

Adult↗

Raw shellfish consumption among renal disease patients. A risk factor for severe Vibrio vulnificus infection.

BACKGROUND: Raw shellfish-associated Vibrio vulnificus septicemia, with a case-fatality rate of nearly 50%, occurs most commonly in immunocompromised patients or those with liver disease. METHODS: Sixty patients with renal disease treated with hemodialysis at The George Washington University and awaiting renal transplantation completed an initial survey that assessed their raw shellfish eating habits and knowledge regarding the pathogen V. vulnificus. Patients were then given educational materials describing the risks of eating raw shellfish and, one month later, completed a second survey that assessed their knowledge retention and intent to eat or not eat raw shellfish in the future. RESULTS: Sixty of 68 (88%) eligible patients completed the survey. Forty-eight percent of patients reported having eaten raw shellfish after being diagnosed with kidney disease, with the highest rates reported among subjects < or = 49 years old and subjects with more than a high school education. Prior to receiving the educational materials, no patient had heard of the pathogen V. vulnificus. Three quarters of patients reported never having been advised by a physician to avoid eating raw shellfish. One month after reading the educational materials, 75% of patients said they would refrain from eating raw shellfish in the future. CONCLUSIONS: In view of their immunocompromised status, patients with end-stage renal disease should be counseled to abstain from eating raw shellfish.

Adolescent↗

A serosurvey of pathogens associated with shellfish: prevalence of antibodies to Vibrio species and Norwalk virus in the Chesapeake Bay region.

Recent concerns regarding the safety of shellfish consumption have focused on the risk posed by naturally occurring marine bacteria such as Vibrio species and by viruses such as Norwalk and related agents. Despite the widespread environmental presence of Vibrio species in the Chesapeake Bay, the rate of reported infections remains low; there have also been no reports of major Norwalk outbreaks associated with shellfish in this area. As infections with these agents may not always be recognized because of difficulties in making the diagnosis and/or their mild or subclinical presentation, a serosurvey was conducted among healthy volunteers living in the Chesapeake Bay region. Serum and questionnaire data were collected during the fall of 1987 from 267 persons with varying levels of exposure to shellfish: shellfish industry workers, persons attending a local seafood festival, and Seventh-day Adventists (who traditionally abstain from eating shellfish). In comparisons among groups, a significant association could not be demonstrated between shellfish consumption or contact and antibody response to Vibrio cholerae O1 or Norwalk virus. Rates of seropositivity were high for both agents (up to 22% seropositive with a V. cholerae O1 Inaba vibriocidal assay, 14% with an enzyme-linked immunosorbent assay for cholera toxin, and up to 70% seropositive with an enzyme-linked immunosorbent assay for antibodies to Norwalk virus); the basis for these responses in population-based studies remains to be determined. Shellfish industry workers did have a significantly elevated antibody response to the unencapsulated phase variant of Vibrio vulnificus as compared with the other groups studied. Infection with V. vulnificus may be relatively common among persons with high levels of exposure to shellfish.

Adult↗

Evaluation of potential indicators of viral contamination in shellfish and their applicability to diverse geographical areas.

The distribution of the concentration of potential indicators of fecal viral pollution in shellfish was analyzed under diverse conditions over 18 months in diverse geographical areas. These microorganisms have been evaluated in relation to contamination by human viral pathogens detected in parallel in the analyzed shellfish samples. Thus, significant shellfish-growing areas from diverse countries in the north and south of Europe (Greece, Spain, Sweden, and the United Kingdom) were defined and studied by analyzing different physicochemical parameters in the water and the levels of Escherichia coli, F-specific RNA bacteriophages, and phages infecting Bacteroides fragilis strain RYC2056 in the shellfish produced, before and after depuration treatments. A total of 475 shellfish samples were studied, and the results were statistically analyzed. According to statistical analysis, the presence of human viruses seems to be related to the presence of all potential indicators in the heavily contaminated areas, where E. coli would probably be suitable as a fecal indicator. The F-RNA phages, which are present in higher numbers in Northern Europe, seem to be significantly related to the presence of viral contamination in shellfish, with a very weak predictive value for hepatitis A virus, human adenovirus, and enterovirus and a stronger one for Norwalk-like virus. However, it is important to note that shellfish produced in A or clean B areas can sporadically contain human viruses even in the absence of E. coli or F-RNA phages. The data presented here will be useful in defining microbiological parameters for improving the sanitary control of shellfish consumed raw or barely cooked.

Animals↗

A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid.

We developed a competitive enzyme-linked immunosorbent assay (ELISA) to analyze brevetoxins, using goat anti-brevetoxin antibodies obtained after immunization with keyhole limpet hemocyanin-brevetoxin conjugates, in combination with a three-step signal amplification process. The procedure, which used secondary biotinylated antibodies, streptavidine-horseradish peroxidase conjugate, and chromogenic enzyme substrate, was useful in reducing nonspecific background signals commonly observed with complex matrices. This competitive ELISA detected brevetoxins in seawater, shellfish extract and homogenate, and mammalian body fluid such as urine and serum without pretreatment, dilution, or purification. We investigated the application of this technique for shellfish monitoring by spiking shellfish meat with brevetoxins and by analyzing oysters from two commercial shellfish beds in Florida that were exposed to a bloom of Karenia brevis (formerly Gymnodinium breve). We performed brevetoxin analysis of shellfish extracts and homogenates by ELISA and compared it with the mouse bioassay and receptor binding assay. The detection limit for brevetoxins in spiked oysters was 2.5 microg/100 g shellfish meat. This assay appears to be a useful tool for neurotoxic shellfish poisoning monitoring in shellfish and seawater, and for mammalian exposure diagnostics, and significantly reduces the time required for analyses.

Animals↗

Quantitation of diarrhetic shellfish poisoning toxins in Chilean mussel using pyrenyldiazomethane as fluorescent labeling reagent.

Diarrhetic shellfish poisoning (DSP) is a gastrointestinal disease caused by lipid soluble polyether toxins produced by dinoflagellates and accumulated in shellfish. Diarrhetic shellfish poisoning is a worldwide threat to public health and the shellfish industry. To date, only four lipid soluble polyethers have been known as diarrhetic shellfish toxins. Among them, Okadaic acid (OA), Dinophysistoxin 1 (DTX-1, 35-methyl OA), Dinophysistoxin 2 (DTX-2, OA isomers) and Dinophysistoxin 3 (DTX-3, 7-O-acyl-35-methyl OA), all of which have free carboxilic groups. To perform quantitative analysis of DSP toxins in shellfish samples is a requirement, because DSP toxins are endemic in the Chilean mollusks of the southern regions, and although human symptoms of DSP appear relatively mild in comparison with the Paralytic Shellfish Poisoning (PSP), the necessity of monitoring the chronic effects of continued uptake of low doses of DSP toxins more closely is imperative, since DSP toxins have been described as potent tumor promoters. This paper shows the synthesis pathway of a chromophore, 1-pyrenyldiazomethane (PDAM), a fluorescent labeling reagent for determination of carboxilic acids, using High Performance Liquid Chromatography with fluorescence on-line detection. This procedure was developed in order to have a quantitative method for DSP toxins analysis that would be useful for health public services and private shellfish industries. The features of this labeling reagent are compared against ADAM and used for quantitative analysis of DSP toxins in Chilean mussels and cultured dinoflagellates samples.

Animals↗

Raw shellfish consumption in California: the 1992 California Behavioral Risk Factor Survey.

We used the 1992 California Behavioral Risk Factor Surveillance System to study the prevalence of raw shellfish consumption in California and the demographic and behavioral characteristics of raw shellfish consumers. We used the logistic regression analysis of the weighted survey data with PC SAS and SUDAAN to adjust for the effects of age and gender. Twenty-three percent of the respondents in the survey reported that they ate raw shellfish; one third of these reported eating raw shellfish once a month. Higher prevalences of raw shellfish consumption were reported by men, persons 18-49 years old, those with income above $25,000 and education beyond high school than by women, individuals older than 49 years, and those with an income of $25,000 or less per year and 12 or fewer years of school. A higher percentage of persons with liver disease, stomach surgery, and a history of chronic alcohol drinking reported consumption of raw shellfish than did individuals without liver disease, previous stomach surgery, or a history of alcohol abuse. After adjustment for gender and age, those who reported acute (P < .01) and chronic (P < .01) drinking and driving while intoxicated (P < .01) were more likely to report consumption of raw shellfish. Two variables (lack of seat belt usage [P = 2] and cigarette smoking [P = .13]) were not significantly associated statistically with raw shellfish consumption.

Adult↗

Improved high-performance liquid chromatographic method for the determination of domoic acid and analogues in shellfish: effect of pH.

Domoic acid (DA) is a naturally-occurring amino acid that causes a form of human intoxication called amnesic shellfish poisoning (ASP) following the consumption of shellfish. A rapid and sensitive HPLC-UV method has been developed for analysis of DA and analogues in shellfish without the need for SPE clean-up. Isocratic chromatographic separation of DA and its isomers from shellfish matrix interferences and from the prevalent amino acid, tryptophan, was achieved by careful control of the mobile phase pH. The optimised pH was found to be 2.5 when using a Luna(2) C18 column. Sample extraction was verified with control extracts from shellfish spiked at 5.0 and 10.0 microg/g of DA and with certified reference material. The average extraction efficiency was 98.5%. The calibration, based on mussel tissue spiked with DA standard, was linear in the range 0.05-5.0 microg/ml (r = 0.9999) and the detection limit (signal:noise 3:1) was better than 25 ng/ml. The DA assay achieved good precision; %RSD = 1.63 (intra-day, n = 6) and %RSD = 3.7 (inter-day, n = 8). This method was successfully applied to a variety of shellfish species, allowing the rapid screening of a large number of samples per day (20-30), without the need for SPE clean-up. Quantitative data were obtained for shellfish samples containing domoic acid in the concentration range 0.25-330 microg/g. Using the same chromatographic conditions, LC-MS3 was used to determine DA and its isomers, isodomoic acid D and epi-domoic acid, in scallop tissues.

Animals↗

Seasonality of diarrhetic shellfish poisoning at a coastal lagoon in Portugal: rainfall patterns and folk wisdom.

Of the three types of toxicity known so far in Portuguese shellfish, only diarrhetic shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) are produced by microalgae that seem to have been present in the last decades or centuries. The most important paralytic shellfish poisoning (PSP) producer, Gymnodinium catenatum, is hypothesised to have been introduced quite recently as only in 1976 PSP toxicity was detected for the first time in shellfish from Galicia, NW Iberian Peninsula. While ASP presents very short episodes of contamination, the concentration of DSP toxins in some years surpasses human safety values for much longer periods. It is traditionally stated that shellfish should be consumed in 'months with R' (September-April). A retrospective study of the maximum monthly DSP levels attained in mussels from a coastal lagoon-Ria de Aveiro-between 1994 and 2001, showed that the highest frequency of months with concentrations surpassing the safety level of 2 microg/g digestive glands were found in June-September, followed by May and October. These months correspond with the months of lowest historical average rainfall in the period 1941-1998. Oscillations in the rainfall pattern coincided with earliest (or latest) detection by HPLC of DSP toxins in mussel in the years studied. In a semi-closed lagunar environment prone to in situ growth of DSP-producer microalgae, like Dinophysis acuminata, rainfall affects river output, lowering salinity and disrupting water column stability that favours Dinophysis growth. The seasonality of DSP recurrence may be connected to the folk adage on safety of shellfish consumption, after many years of empirical observations by coastal populations of diarrhoea episodes in summertime.

Animals↗

Metabolic transformation of dinophysistoxin-3 into dinophysistoxin-1 causes human intoxication by consumption of O-acyl-derivatives dinophysistoxins contaminated shellfish.

This paper describes for the first time a massive intoxication episode due to consumption of shellfish contaminated with 7-O-acyl-derivative dinophysistoxin-1, named Dinophysistoxin-3 (DTX-3). 7-O-acyl-derivative dinophysistoxin-1, a compound recently described in the literature, was found in shellfish samples collected in the Chilean Patagonia fjords. This compound does not inhibit Protein Phosphatases and also does not elicit the symptoms described for Diarrheic Shellfish Poisoning (DSP). The data showed here, give evidence of metabolic transformation of 7-O-acyl-derivative dinophysistoxin-1 (DTX-3) into Dinophysistoxin-1 (DTX-1, Methyl-Okadaic acid) in intoxicated patients. This metabolic transformation is responsible for the diarrheic symptoms and the intoxication syndrome showed by patients that consumed contaminated shellfish, which showed only the presence of 7-O-acyl-derivative dinophysistoxin-1. Patients fecal bacterial analysis for the presence of enteropathogens was negative and the mouse bioassay for DSP, performed as described for regulatory testing, was also negative. The HPLC-FLD and HPLC-MS analysis showed only the presence of DTX-3 as the only compound associated to DSP toxins in the contaminated shellfish samples. No other DSP toxins were found in the shellfish sample extracts. However, the patient fecal samples showed DTX-1 as the only DSP toxins detected in fecal. Moreover, the patient fecal samples did not show DTX-3. Since 7-O-acyl-derivative dinophysistoxin-1 (DTX-3) was the only compound associated to DSP toxins detected in the shellfish samples, an explanation for the diarrheic symptoms in the intoxicated patients would be the metabolic transformation of DTX-3 into DTX-1. This transformation should occur in the stomach of the poisoned patients after consuming 7-O-acyl-derivatives dinophysistoxin-1 (DTX-3) contaminated bivalves.

Abdominal Pain↗

[Determination of saxitoxin in canned shellfish (author's transl)].

Poisonings by saxitoxin-containing shellfish occur regularly in shore areas. The reason is increased growth of the dinoflagellates Gonyaulax tamarensis and Gonyaulax catenella. Due to the widespread consumption of canned shellfish these kinds of poisoning also occurs in continental areas. Therefore it is necessary to determine saxitoxin in canned shellfish products. Because of their sensitivity fluorospectrophotometric determinations of saxitoxin are preferred. However, the methods described in the literature can only be applied to fresh shellfish. Consequently a method for the determination of saxitoxin in canned shellfish was developed. This method offers the advantage that parallel with the fluorophotometric determination a biotest with mice can be carried out with the same extract for forensic corroboration of the results. The extent of saxitoxin occurence in Spanish canned shellfish in Austria in the years 1976-1979 is described. Apparently the producers of canned shellfish were able to solve this problem since mid - 1978.

Animals↗