Radionuclides in Washington shellfish, January 1963-December 1967.
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Diarrhetic shellfish poisoning (DSP) is a gastrointestinal disease caused by fat-soluble polyether toxins produced by dinoflagellates and accumulated in shellfish. Up to the present, only four fat-soluble polyethers have been known as diarrhetic shellfish toxins. Among them,*** Okadaic acid, Dinophysistoxin-1, Dinophysistoxin-2 and Dinophysistoxin-3. Outbreaks associated with DSP have occurred in the Chilean Patagonia fjords since 1970. Native people, who live in small communities close to the southern fjords, smoke fresh shellfish. During this popular smoking procedure, they impregnate the shellfish with polycyclic aromatic hydrocarbons, as incomplete combustion products, which are potent carcinogenic compounds, this product is sold in local markets without phycotoxins analysis or inclusion in any monitoring program. The present paper shows, DSP phycotoxins quantitation, using high performance liquid chromatography with fluorescent and mass spectrometric detection and the measurements of polycyclic aromatic hydrocarbons by gas chromatography with mass detection, in smoked shellfish samples. The presence of Dinophysistoxin-3, the Dinophysistoxin-1 ester (7-O-acyl-derivatives of dinophysistoxin-1), was assessed in all shellfish samples analyzed. The 7-OH in Dinophysistoxin-1 was esterified with palmitic fatty acid. The shellfish meat contains seven polycyclic aromatic hydrocarbons, among them fluoranthene, phenanthrene, anthracene, pyrene and benzo[a]pyrene. The gas chromatography-mass spectrometry analysis showed four of the six most frequent carcinogenic polycyclic aromatic hydrocarbons reported. The content of benzo[a]pyrene in the Razor Clam and Ribbed Mussel were 78.61 and 4.94 ng/g of shellfish dry weight, respectively. In both cases the benzo[a]pyrene amounts were greater than the acceptable tolerance limits of 1 microg/kg of sample. The Razor Clam samples also show amount further above the maximum label regulated by FAO/WHO (10 microg/kg). The presence of both type of compounds in the smoked shellfish samples analyzed, correspond to a dangerous combination, where the polycyclic aromatic hydrocarbons are carcinogenic compounds by themselves and DTX-1, is a potent tumor promoter.
In the United States of America, seafood ranked third on the list of products which caused foodborne disease between 1983 and 1992. Outbreaks connected with fish vectors were caused by scombroid, ciguatoxin, bacteria and unknown agents; in shellfish, unknown agents, paralytic shellfish poisoning, Vibrio spp. and other bacteria, followed by hepatitis A virus, were responsible for the outbreaks. At least ten genera of bacterial pathogens have been implicated in seafood-borne diseases. Over the past twenty-five years, bacterial pathogens associated with faecal contamination have represented only 4% of the shellfish-associated outbreaks, while naturally-occurring bacteria accounted for 20% of shellfish-related illnesses and 99% of the deaths. Most of these indigenous bacteria fall into the family Vibrionaceae which includes the genera Vibrio, Aeromonas and Plesiomonas. In general, Vibrio spp. are not associated with faecal contamination and therefore faecal indicators do not correlate with the presence of Vibrio. Viruses are the most significant cause of shellfish-associated disease: in New York State, for example, 33% and 62% of 196 outbreaks between 1981 and 1992 were caused by Norwalk virus and gastrointestinal viruses (small round structured viruses), respectively. In addition, several illnesses are a result of toxic algal blooms, the growth of naturally occurring bacteria and diatoms causing neurotoxic shellfish poisoning, paralytic shellfish poisoning, diarrhoetic shellfish poisoning, amnesic shellfish poisoning and ciguatera. Current estimates place the annual number of ciguatera cases at 20,000 world-wide. Scombroid poisoning is the most significant cause of illness associated with seafood. Scombrotoxin is of bacterial origin and halophilic Vibrio spp. causing high histamine levels are implicated as the source. Scombroid poisoning is geographically diverse and many species have been implicated, namely: tuna, mahi-mahi, bluefish, sardines, mackerel, amberjack and abalone. Temperature abuse has been cited as a major cause of scombroid poisoning. For routine work, the use of faecal indicators to predict the relative level of faecal contamination should not be disposed of. However, the main source of seafood illness is due to species which are not predicted by these organisms. In order to protect public health, routine surveillance using new pathogen-specific techniques such as polymerase chain reaction should be used. This, in combination with risk assessment methods and hazard analysis and critical control points, will begin to address the need for improvement in the safety of seafood.
Domoic acid (DA) was first detected in shellfish in New Zealand after the implementation of a comprehensive biotoxin monitoring programme for amnesic, paralytic, diarrhetic and neurotoxic shellfish toxins, following a suspected neurotoxic shellfish poisoning (NSP) event in early 1993. Both phytoplankton monitoring and shellfish flesh testing programmes have led to an extensive database which has helped link species of Pseudo-nitzschia to specific DA outbreaks. In 1994, P. pungens and P. turgidula were associated with DA contamination of shellfish, and cultured isolates of these species proved to be toxin producers. During 1996 the use of species-specific ribosomal RNA (rRNA)-targeted oligonucleotide probes and DA immunoassays led to the discovery of toxin production by P. fraudulenta, and showed the nontoxic P. heimii to be a major bloom former. Pseudo-nitzschia delicatissima, P. pseudodelicatissima and P. multiseries, also identified using rRNA-targeted probes, have been linked to DA contamination of New Zealand shellfish; P. australis is the main cause of DA in scallops. The relative amnesic shellfish poisoning (ASP) risk associated with different species, largely determined by DA immunoassays of cultured isolates, is now used by some regulators to refine risk assessments. Species identification is therefore vital so that shellfish growers, and health and industry officials, can make safe and economically sound harvesting decisions. The development and field trialling of DNA probes is proving invaluable in this context.
During May and June 1994, the authors interviewed and requested a shellfish sample from a population-based sample of 170 residents from Kodiak and Old Harbor, Alaska. Of 51 Old Harbor and 68 Kodiak residents who had eaten shellfish gathered from Kodiak Island, 18 and 6 percent, respectively, had a history of PSP. We calculated the incidence of paralytic shellfish poisoning in Old Harbor and Kodiak as 15 and 1.5 per 1000 persons per year, respectively. Of 12 butter clam batches collected from residents, 6 had a paralytic shellfish poison toxin level greater than the regulatory limit of 80 micrograms saxitoxin equivalent per 100 g of tissue; one of the 29 people who ate these shellfish developed illness. People who eat shellfish collected from non-commercial beaches have a high rate of paralytic shellfish poisoning. It may be possible to raise the regulatory level for paralytic shellfish poison toxin without affecting the public health.
Because shellfish (oysters, clams, and mussels) are filter-feeders, pathogens become concentrated within them, and human consumption of raw, or under-cooked shellfish can result in disease outbreaks. Identification of hepatitis A virus (HAV) in shellfish has been difficult for several reasons: the concentration of virions in shellfish tissues are very low, detection methods based on in vitro propagation are unreliable, recovery of virions from shellfish tissues is inefficient, and PCR inhibitors in shellfish tissues limit the success of RT-PCR. These facts underlie difficulties in determining cause and effect relationships between hepatitis A outbreaks and detection of HAV contamination in shellfish samples. We have developed a reliable and highly sensitive method for detection of HAV in oyster tissues at low levels (0.001 FFU/ml-fluorescent focus units per milliliter). Our method combines dissection of the gastrointestinal oyster tract, organic extraction before PEG precipitation, and RNA extraction with Trizol LS, followed by RT-PCR and hybridization using a digoxigenin-labeled HAV cDNA probe. Our results will benefit both public health officials concerned about hepatitis A infections caused by consumption of HAV-contaminated oysters and shellfish producers who require reliable methods for quality control of commercial oyster production.
A study of the presence of hepatitis A virus (HAV) and enterovirus (EV) in shellfish from the northwestern coast of Spain, one of the most important mussel producers in the world, was carried out employing dot-blot hybridization and RT-PCR techniques. In addition, bacterial contamination of the samples was evaluated by Escherichia coli (EC) counts, according to the European Union (EU) standards of shellfish microbiological quality. Shellfish samples included raft-cultured and wild mussels, as well as wild clams and cockles. Bacterial counts showed that the majority of samples (40.8%) could be classified as moderately polluted following the EU standards, and therefore should undergo depuration processes. However, differences in bacterial contamination were observed between cultured mussel and wild shellfish. Thus, percentage of clean samples (<230 EC/100 g shellfish) was clearly higher in cultured mussels (49.1%) than in wild mussels (22.8%) or clams and cockles (10.7%). HAV was detected in 27.4% and EV in 43.9% of the samples that were analyzed. Simultaneous detection of both viral types occurred in 14.1% of the samples. Statistical tests of dependence (chi-square test) showed no relationship either between viral and bacterial contamination, or between the presence of HAV and EV. Comparative analysis of hybridization and RT-PCR for viral detection yielded different results depending on the virus type that was studied, RT-PCR being effective for HAV but not for EV detection. The obtained results reinforce once again the inadequacy of bacteriological standards to assess viral contamination and suggest that although virological analysis of shellfish is possible by molecular techniques, interlaboratory standardization and validation studies are needed before the routine use in monitoring shellfish microbiological safety.
Gastroenteritis outbreaks linked to shellfish consumption are numerous and Norwalk-like viruses (NLVs) are frequently the responsible causative agents. However, molecular data linking shellfish and clinical samples are still rare despite the availability of diagnostic methods. In a recent outbreak we found the same NLV sequence in stool and shellfish samples (100% identity over 313 bp in the capsid region), supporting the epidemiological data implicating the shellfish as the source of infection. A semiquantitative approach using most-probable-number-RT-PCR (MPN-RT-PCR) demonstrated the presence of a hundred of RT-PCR units per oyster. Follow-up of the oysters in the harvest area, for approximately 2 months, showed persistence of NLV contamination of the shellfish at levels up to a thousand RT-PCR units per oyster prior to depuration of the shellfish. This finding is useful in beginning to understand shellfish contamination and depuration for use in future hazard analyses.
AIMS: To examine the relationships among increasing estuarine shellfish closings due to bacterial contamination, adjacent shoreline land uses and environmental variables. METHODS AND RESULTS: A 1 year study of faecal coliform bacterial contamination of a small estuary in central NC, USA was done relative to adjacent land uses. The area has experienced rapid growth in residential shoreline development including the installation of adjacent, separate docking facilities for larger boats, each <11 slips (pseudomarina) that appear to be a single marina (individual facilities of >10 slips). Six near-shore sites were selected [old developed shore (OD), undeveloped shore (UD), two pseudomarinas (P1, P2), newly developed shore (ND) and a real marina (RM)]. Five locations were spaced along the shore near each site. Paired Thursday/Monday samples were collected biweekly (summer) and monthly (other seasons). Results indicate that OD had the highest bacteria counts followed by ND, RM and P1 & P2. Three sites (OD, ND and RM) failed to meet NC shellfishing waters standards at all locations. At the pseudomarina sites 4 of 10 locations failed to meet shellfish standards while two locations at UD failed to meet these standards. There were no significant differences between paired Thursday/Monday samples. At three sites (OD, UD and P2) bacteria counts were positively correlated with increased water level due to wind tides. CONCLUSIONS: Any type of estuarine shoreline development may result in closing of adjacent shellfishing waters. ND had bacterial counts second only to OD in spite of the retention of vegetated shoreline buffers and very new septic systems. As expected, the RM also failed to meet shellfish standards. Unexpectedly, only four of the 10 pseudomarina locations failed to meet the standards. Weekend boat use had no effect on bacterial counts. Surface runoff from rain and shoreline flooding from increased water levels increased bacterial counts, probably as a result of suspension of surface deposited faeces from wildlife and domestic animals. SIGNIFICANCE AND IMPACT OF THE STUDY: Multiple docking facilities do not necessarily result in violations of shellfish water quality standards. However, the elevated bacterial counts observed along the newly developed shore suggest caution in approving the practice of allowing individual 'oyster gardening' off private piers if the oysters are intended for human consumption. The practice of automatic closure of shellfish waters around RMs was supported. Correlations of bacterial counts with time following significant rainfall suggests a sampling strategy to separate local sources of bacteria from more remote sources thus focusing limited remedial resources more effectively.
Infective Cryptosporidium parvum oocysts were detected in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) from a shellfish-producing region (Gallaecia, northwest Spain, bounded by the Atlantic Ocean) that accounts for the majority of European shellfish production. Shellfish were collected from bay sites with different degrees of organic pollution. Shellfish harboring C. parvum oocysts were recovered only from areas located near the mouths of rivers with a high density of grazing ruminants on their banks. An approximation of the parasite load of shellfish collected in positive sites indicated that each shellfish transported more than 10(3) oocysts. Recovered oocysts were infectious for neonatal mice, and PCR-restriction fragment length polymorphism analysis demonstrated a profile similar to that described for genotype C or 2 of the parasite. These results demonstrate that mussels and cockles could act as a reservoir of C. parvum infection for humans. Moreover, estuarine shellfish could be used as an indicator of river water contamination.
Noroviruses (called formerly "Norwalk-like viruses") cause food-borne gastroenteritis outbreaks. These outbreaks were thought to be related to shellfish consumption, although non-shellfish related outbreaks also occurred frequently in Japan. To clarify the epidemiology of Norovirus outbreaks, 435 stool samples were collected from 60 acute non-bacterial gastroenteritis outbreaks occurring over 8 years in Okayama, Japan. Using reverse transcription-PCR (RT-PCR), Noroviruses were detected in 257 cases (59.1% of all samples) from 46 outbreaks (77% of all outbreaks). The majority of the 46 Norovirus outbreaks (89%) occurred during November to March; notably one-third occurred in December. Restaurants, schools, and welfare institutions accounted for the major settings in 50%, 20%, and 15% of the Norovirus outbreaks, respectively. This was similar to other reports from Japan, but differed from those from the United Kingdom. The transmission routes were assigned in 27 of the Norovirus outbreaks. In 18 outbreaks the routes were related to human contact (7 from food handlers and 11 from person-to-person contact), whereas those related directly to shellfish occurred only in 9 outbreaks. These results suggest that transmission routes related to human contact are more important than recognized previously in the context of preventive medicine. Furthermore, all outbreaks in which some of the samples contained dual genogroups of Noroviruses were related to shellfish, suggesting that consumption of contaminated shellfish frequently results in mixed Norovirus infections in contrast to other transmission routes and that coexistence of genogroups is a useful marker for shellfish-related outbreaks.
National standards and guidelines for pesticides can be useful tools in water-quality assessment for evaluating potential human health or ecological effects of measured pesticide residues in water, bed sediment, or aquatic organisms. However, valid use of a given standard or guideline requires an understanding of its technical basis and underlying assumptions. Each type of standard or guideline is specific for one sampling medium (water, bed sediment, and fish and shellfish tissue) and is aimed at protection of one or more beneficial uses of the hydrologic system (drinking water, fish and shellfish consumption, aquatic organisms, and wildlife). These characteristics can be used to identify which standards and guidelines are appropriate for comparison with measured pesticide concentrations in environmental samples from a given hydrologic system. A review of standards and guidelines can be restricted to the applicable sampling medium. Then, the beneficial uses of the hydrologic system need to be identified and the measured pesticide concentrations compared with standards and guidelines for all beneficial uses that apply to that system. Several key factors that must be considered when applying this general process to water-quality assessment are summarized below. Two precautions need to be considered regarding sampling media: 1. Standards and guidelines for water distinguish between finished drinking water (potable water, often treated) and ambient surface water. If standards and guidelines for drinking water (EPA primary drinking-water regulations and drinking-water health advisories) are applied to measured pesticide concentrations in ambient water samples, the effects of water treatment (such as filtration) need to be considered. 2. Standards and guidelines for fish and shellfish tissue distinguish between edible fish and shellfish tissue and whole fish tissue. Comparison of pesticide concentrations in whole fish tissue with standards or guidelines for edible fish and shellfish tissue is appropriate only as a screening procedure to determine whether additional sampling and analysis for contaminants in edible fish fillets are warranted. For some sampling media (water, fish and shellfish tissues), both standards and guidelines may exist for a given pesticide. Standards and guidelines may differ in their technical bases and in the implications or consequences of finding measured concentrations in exceedance of the standard or guideline value. Therefore, comparison of measured pesticide concentrations with both standards and guidelines is useful because each provides different information about the hydrologic system.(ABSTRACT TRUNCATED AT 400 WORDS)
A randomized telephone survey of 3015 women was conducted in an effort to assess the effectiveness of local sport-fish consumption advisories. Survey participants were between the ages of 18 and 45 and lived in the states of Arkansas, California, Connecticut, Florida, Maine, Minnesota, Montana, New Jersey, New Mexico, North Carolina, and Wisconsin. At the time of the women's interview, fish and shellfish consumption information was obtained for children under 18 years of age living in the household. One child (aged 2-17) from each household (1852) was randomly selected to evaluate fish consumption among children. Based on maternal recall, 84% of these children had consumed fish or shellfish at least once during the previous 12 months. This percentage ranged from 73% in New Jersey to 94% in Louisiana and was higher among children who lived with a licensed angler compared to those who did not. Eight percent of the children ate fish and/or shellfish more than twice a week. Of the total number of fish and shellfish meals eaten by children, 67% was commercial finfish, 22% was shellfish, and 11% was sport-caught finfish. Among those who ate fish, the average consumption rate was 47 meals per year-slightly less than one meal per week. This consumption frequency rate varied by state of residence ranging from 37 meals per year in Montana and Wisconsin to 62 in Florida. Because of these regional differences, the use of national average fish consumption rates may over- or under-estimate consumption in localized areas. This survey suggests that targeting information to women who eat fish may also protect children; more than 80% of children have fish consumption patterns that are similar to that of their mothers. Additional research and biomonitoring is needed to improve our understanding of the risk and benefits associated with childhood consumption of fish and shellfish.
1. The present paper reviews the toxic dinoflagellates found in Singapore waters that produce toxins that can accumulate through marine food chains to cause seafood poisonings. 2. Singapore waters contain dinoflagellate species linked to three types of seafood poisoning: paralytic shellfish poisoning, diarrhetic shellfish poisoning (DSP) and ciguatera. 3. Paralytic shellfish poisoning and DSP occur by eating bivalve shellfish contaminated with saxitoxins and okadaic acid analogues, respectively. Shellfish accumulate these toxins from filter feeding on a number of species of (mostly) planktonic dinoflagellates. 4. In contrast, benthic species of dinoflagellates of the genus Gambierdiscus produce the ciguatoxins that are bioaccumulated into finfish to cause ciguatera. 5. Paralytic shellfish poisoning and DSP are the major concern for local and regionally produced seafood. To the best of our knowledge, ciguatera poisoning in Singapore only originates from imported reef fish.
Seafood, including fish, shrimp, lobster, crab, crayfish, mussel, and clam are among the most frequent causes of food allergy. Seafood poisoning, including reactions to natural toxins, frequently masquerades as an allergic reaction on presentation. Ingestion of contaminated shellfish results in a wide variety of symptoms, depending on the toxins present, their concentrations in the shellfish, and the amount of contaminated shellfish consumed. Five types of shellfish poisoning have been identified clearly including paralytic, neurotoxic, diarrhetic, amnestic, and azaspiracid shellfish poisonings. Based on the presence or absence of the toxin at the time of capture, fish poisoning can be considered conceptually in two categories. In ciguatera and puffer fish poisoning, the toxin is present in live fish, whereas in scombroid, it is produced only after capture, in the fish flesh, by contaminating bacteria because of improper refrigeration. Most shellfish-associated illness is infectious in nature (bacterial or viral), with the Norwalk virus accounting for most cases of gastroenteritis.
Toxin profiles were determined in phytoplankton cell concentrates and Greenshell mussels (Perna canaliculus) exposed to a dinoflagellate bloom dominated by Dinophysis acuta and Protoceratium reticulatum. This was achieved by using a method for the simultaneous identification and quantification of a variety of micro-algal toxins by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionisation (+/-) and monitoring of daughter ions in multiple reaction modes. Plankton concentrates and shellfish contained high levels of yessotoxins (YTXs) and pectenotoxins (PTXs) and low levels of okadaic acid (OA). A high proportion (>87%) of the OA in both plankton and shellfish was released by alkaline hydrolysis. An isomer of pectenotoxin 1 (PTX1i) was nearly as abundant as pectenotoxin 2 (PTX2) in the plankton and shellfish, and the latter contained high levels of their respective seco acids. DTX1, DTX2, and PTX6 were not detected. MS-MS experiments revealed that the shellfish contained several other oxygenated metabolites of YTX in addition to 45-hydroxy yessotoxin (45OH-YTX). Gymnodimine (GYM) was present in the shellfish but not plankton and it was probably the residue from a previous GYM contamination event. Unlike the other toxins, GYM was concentrated in tissues outside the digestive gland and levels did not decrease over 5 months. The depuration rates of YTX and PTXs from mussels were modelled.