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Evaluation of 11-[3H]-tetrodotoxin use in a heterologous receptor binding assay for PSP toxins.

This report describes the preparative scale production of 11-[3H]-tetrodotoxin (TTX) and its evaluation as a substitute for [3H]-saxitoxin (STX) as the radioligand in a receptor binding assay for paralytic shellfish poisoning (PSP) toxins. Restrictions on the world-wide distribution of [3H]-STX imposed by the international Chemical Weapons Convention served as the primary impetus for this study. We have incorporated on a preparative scale, a nonexchangeable tritium label into the TTX molecule at a specific activity of 12.90 Ci/mmol and recovered material of high radiochemical purity (98%). The resulting 11-[3H]-TTX was found to exhibit site-specific binding characteristics in the receptor assay (dissociation constant(K(d))=4.77+/-1.54nM; maximum binding(B(max))=1. 62+/-0.24pmol/mg of synaptosomal protein). The inhibition constant (K(i)) for the assay was 1.46+/-0.28 nM STX equiv. (n=6), with an estimated detection limit of ca. 2-4 ng STX equiv./ml in a sample extract. Moreover, quantitative comparisons indicated that 11-[3H]-TTX could be used interchangeably with [3H]-STX in the receptor assay for determination of PSP toxicity in shellfish and algal extracts without compromising assay performance. We conclude that the 11-[3H]-TTX produced and evaluated herein exhibits physical, chemical and biological characteristics suitable not only for use in the PSP receptor binding assay, but likely for other applications employing [3H]-STX as the radioligand.

Evaluation Studies as Topic↗

Quadrupole ion storage tandem mass spectrometry and high-resolution mass spectrometry: complementary application in the measurement of 2,3,7,8-chlorine substituted dibenzo-p-dioxins and dibenzofurans in US foods.

The US Food and Drug Administration has simultaneously utilized both high-resolution mass spectrometry (HRMS) and quadrupole ion storage tandem mass spectrometry (QISTMS) in the measurement of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) in 147 food samples collected in 1998 and 1999 in the US. In 1998, 20 egg samples, six scallop, 10 blue crab, eight American lobster, 10 pollack, 15 striped bass, five rockfish, 10 crawfish, seven aqua-cultured and 13 wild-caught salmon, along with 19 cream and 18 mozzarella cheese samples were measured for PCDD/Fs. QISTMS provided limits of detection (LODs) close to those produced using HRMS for many congeners in 56 samples analyzed by both techniques in 1998 and three salmon and three striped bass collected in 1999. The I-TEQs of the mean levels for measured congeners in 40 samples of fish and shellfish and 16 cheese and eggs from 1998 analyzed by HRMS and QISTMS were 0.99 and 1.1 ng/kg wet weight, respectively. The I-TEQ for mean congener levels in the 40 fish and shellfish measured by HRMS was 1.4 ng/kg wet weight. A higher sample throughput with greater data quality at a lower cost is achievable by using both QISTMS and HRMS.

Animals↗

Screening-level ecological risk assessment of polychlorinated dibenzo-p-dioxins and dibenzofurans in sediments and aquatic biota from the Venice Lagoon, Italy.

Recent monitoring data indicate that portions of Italy's Venice Lagoon ecosystem have been degraded due to biological and chemical pollution from a variety of potential sources. Using polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) data collected from sediment, fish and shellfish in the Lagoon, a screening-level ecological risk assessment (ERA) was performed to evaluate the risks to representative aquatic biota and wildlife receptors. Risks to aquatic invertebrates posed by PCDD/Fs in sediment were evaluated by comparing measured tissue concentrations in fish and shellfish to appropriate ecotoxicological reference values. For mammalian and avian receptors, risks posed by theoretical exposures to PCDD/Fs through the food chain were calculated using conservative wildlife exposure models. Results of the screening-level approach indicate that the potential for adverse effects to fish and aquatic invertebrate receptors from PCDD/Fs in surficial sediments are unlikely. Adverse effects to wildlife are possible but highly uncertain, and warrants further investigation in a more comprehensive ERA.

Animals↗

Contamination of bivalve molluscs by Cryptosporidium oocysts: the need for new quality control standards.

A yearlong study was carried out to investigate the presence and viability of Cryptosporidium oocysts in 203 samples of cultured shellfish from Galicia (NW Spain) and 38 samples imported from other European Union (EU) countries. Shellfish samples included mussels, oysters, clams and cockles. Cryptosporidium oocysts were detected, using a direct immunofluorescence antibody test (IFAT), in 34.4% of the samples analyzed; use of the fluorogenic dye propidium iodide (PI) revealed viable potentially infective oocysts in 53.0% of these samples. There was no relation between the presence of Cryptosporidium oocysts and the microbiological contamination detected in the samples expressed as Most-Probable-Number (MPN) of fecal coliforms, the different species of mollusc, or the month of sampling. One important finding was that the depuration process was ineffective in totally removing oocyst contamination. Furthermore, the existence of viable oocysts in samples with microbiological contamination levels lower than 300 fecal coliforms/100 g, which in accordance with current legislation are considered suitable for human consumption, suggests the need to include parasitological analyses in the quality control for these molluscs.

Animals↗

Selective surface adhesion of the toxic microalga Alexandrium minutum induced by contact with substituted polystyrene derivatives.

On the basis of observations that biospecific random copolymers (RACS) could induce phenotypic changes on contact with selected eukaryotic or prokaryotic cell lines, polystyrene derivatives of known compositions and obtained by random substitutions of sodium sulfonate and of sulfamides of aspartic acid dimethyl ester, phenylalanine and leucine, were placed in contact with swimming dinophytes of the PSP toxin producing species Alexandrium minutum and of the non-toxic species Heterocapsa triquetra. A. minutum cells exhibited higher adhesion for the random copolymer made up of polystyrene (29%), polystyrene aspartic acid dimethyl ester sulfamide (47%) and polystyrene sodium sulfonate (24%), than for samples of this series with different compositions. In contrast to this, A. minutum adhesion remained very low throughout the phenylalanine and leucine copolymer series. These results indicate that the cell-substrate adhesion phenomenon is dependent upon the final composition of the copolymer, i.e. that it is composition-specific. Taxonomic specificity was then demonstrated by presenting the PSAspOMe copolymer series with cells of the non toxic species H. triquetra (Peridinialia) related to A. minutum (Gonyaulacacea), and by observing no specific association, i.e. no signal above background levels at any composition. Specific ligand-cell adhesion is evidenced for the first time between biospecific RACS and phytoplankton, which may inspire a new generation of structures to be used in aquaculture as protective nets over shellfish clusters, or as selective filtering devices to assist in shellfish depuration from toxic microalgae.

Animals↗

Gas chromatographic-mass spectrometric determination of 4-nonylphenols and 4-tert-octylphenol in biological samples.

A simple and rapid method is described for the GC-MS determination of 4-nonylphenols (NOs) and 4-tert-octylphenol (OC) in biological samples. The NOs and OC in the sample are extracted with acetonitrile and the lipid in the sample extract is eliminated by partitioning between hexane and acetonitrile. After Florisil PR column clean-up, the sample extract is analyzed by GC-MS in the selected ion monitoring (SIM) mode. Average recoveries in pale chub (fish) and corbicula (shellfish) are 86.0 and 93.4% for NOs, and 95.8 and 96.4% for OC, respectively, spiked at the levels of 1.0 microg of NOs and 0.1 microg of OC per 5 g of fish and shellfish samples. The detection limits are 20 ng/g for NOs and 2 ng/g for OC.

Animals↗

A comparison of strategies for the detection and recovery of Vibrio vulnificus from marine samples of the western Mediterranean coast.

We have compared the effectiveness of culture-based methods and a DNA-based method for the detection, of Vibrio vulnificus from a seawater and three types of shellfish collected from the costal waters of Valencia, Spain. For culture-based method, we used two selective media, thiosulphate-citrate-salts-sucrose (TCBS), and cellobiose-polymyxin B-colistin (CPC) agars with and without previous enrichment in alkaline-saline-peptone-water (APWS). Presumptive colonies were confirmed as V. vulnificus by the polymerase chain reaction (PCR) using previously described 23S rRNA V. vulficus-specific sequences as primers (Dvu 9V and Dvu 45R). Direct detection was accomplished by a nested-PCR procedure developed for environmental samples, with the above mentioned primers for the second amplification. Of 32 seawater samples, only one yielded positive results by direct detection by PCR, whereas five were positive by culture methods. Of the 32 bivalve samples, two were positive by PCR and five by culture methods. From a total of 675 presumptive colonies selected on the two media, only 48 (20 from seawater and 28 from bivalves) were confirmed as V. vulnificus by PCR. Forty-six V. vulnificus isolates were obtained after enrichment and only two after direct inoculation of CPC. Except for one sampling, positive results by direct detection did not correlate with confirmed strains obtained from culture media. API 20E profiles were recorded for all isolates previously identified as V. vulnificus, revealing that around 20% of the strains were sucrose-positive. For our samples, the best strategy consisted in the combination of culture based methods (3 h enrichment in APWs at 40 degrees C followed by CPC at the same temperature) and DNA-based procedures (specific PCR amplification of the presumptive colonies with primers Dvu 9V and Dvu 45 R), which allowed the detection and accurate identification of V. vulnificus in less than 48h. This is the first report on the detection of cells of V. vulnificus naturally present in seawater and edible shellfish in the Spanish Mediterranean coast.

Animals↗

Brain-specific lipids from marine, lacustrine, or terrestrial food resources: potential impact on early African Homo sapiens.

The polyunsaturated fatty acid (PUFA) composition of the mammalian central nervous system is almost wholly composed of two long-chain polyunsaturated fatty acids (LC-PUFA), docosahexaenoic acid (DHA) and arachidonic acid (AA). PUFA are dietarily essential, thus normal infant/neonatal brain, intellectual growth and development cannot be accomplished if they are deficient during pregnancy and lactation. Uniquely in the human species, the fetal brain consumes 70% of the energy delivered to it by mother. DHA and AA are needed to construct placental and fetal tissues for cell membrane growth, structure and function. Contemporary evidence shows that the maternal circulation is depleted of AA and DHA during fetal growth. Sustaining normal adult human brain function also requires LC-PUFA.Homo sapiens is unlikely to have evolved a large, complex, metabolically expensive brain in an environment which did not provide abundant dietary LC-PUFA. Conversion of 18-carbon PUFA from vegetation to AA and DHA is considered quantitatively insufficient due to a combination of high rates of PUFA oxidation for energy, inefficient and rate limited enzymatic conversion and substrate recycling. The littoral marine and lacustrine food chains provide consistently greater amounts of pre-formed LC-PUFA than the terrestrial food chain. Dietary levels of DHA are 2.5-100 fold higher for equivalent weights of marine fish or shellfish vs. lean or fat terrestrial meats. Mammalian brain tissue and bird egg yolks, especially from marine birds, are the richest terrestrial sources of LC-PUFA. However, land animal adipose fats have been linked to vascular disease and mental ill-health, whereas marine lipids have been demonstrated to be protective. At South African Capesites, large shell middens and fish remains are associated with evidence for some of the earliest modern humans. Cape sites dating from 100 to 18 kya cluster within 200 km of the present coast. Evidence of early H. sapiens is also found around the Rift Valley lakes and up the Nile Corridor into the Middle East; in some cases there is an association with the use of littoral resources. Exploitation of river, estuarine, stranded and spawning fish, shellfish and sea bird nestlings and eggs by Homo could have provided essential dietary LC-PUFA for men, women, and children without requiring organized hunting/fishing, or sophisticated social behavior. It is however, predictable from the present evidence that exploitation of this food resource would have provided the advantage in multi-generational brain development which would have made possible the advent of H. sapiens. Restriction to land based foods as postulated by the savannah and other hypotheses would have led to degeneration of the brain and vascular system as happened without exception in all other land based apes and mammals as they evolved larger bodies.

Animals↗

Neurotoxic marine poisoning.

Marine poisoning results from the ingestion of marine animals that contain toxic substances and causes substantial illness in coastal regions. Three main clinical syndromes of marine poisoning have important neurological symptoms-ciguatera, tetrodotoxin poisoning, and paralytic shellfish poisoning. Ciguatera is the commonest syndrome of marine poisoning and is characterised by moderate to severe gastrointestinal effects (vomiting, diarrhoea, and abdominal cramps) and neurological effects (myalgia, paraesthesia, cold allodynia, and ataxia), but is rarely lethal. Tetrodotoxin poisoning and paralytic shellfish poisoning are less common but have a higher fatality rate than ciguatera. Mild gastrointestinal effects and a descending paralysis are characteristic of these types of poisoning. In severe poisoning, paralysis rapidly progresses to respiratory failure. Diagnosis of all types of marine poisoning is made from the circumstances of ingestion (type of fish and location) and the clinical effects. Because there are no antidotes, supportive care, including mechanical ventilation in patients with severe paralysis, is the mainstay of treatment.

Animals↗

The distribution of Vibrio parahaemolyticus in British coastal waters: report of a collaborative study 1975--6.

A collaborative survey has shown that V. parahaemolyticus is widely distributed in British coastal waters, sediments and shellfish, especially in southern and western areas. The relatively small numbers in the environment do not present significant health hazards from marine products, provided that processing, storage and distribution are adequate. The presence of this organism in small numbers in British coastal waters or in shellfish should not in itself be regarded as cause for concern.

Mollusca↗

Polybrominated diphenyl ethers (PBDEs) in foodstuffs: human exposure through the diet.

Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in a variety of materials, including synthetic polymers and textiles. Although these chemicals have been detected in environmental samples and human tissues, there is little information about human exposure to PBDEs through the diet. In the present study, we determined the concentrations of PBDEs in a number of food samples acquired in Catalonia (Spain) during 2000. The dietary intake of PBDEs was estimated for the general population living in this Spanish region. The highest PBDE concentrations were found in oils and fats, fish and shellfish, meat and meat products, and eggs, while the lowest levels corresponded to fruits, vegetables, and tubers. The dietary intake of PBDEs for an adult male was 97.3 ng/day (assuming not detected (ND) = (1)/(2) limit of detection (LOD)) or 81.9 ng/day (assuming ND = 0) The greatest contribution to these values corresponded to fish and shellfish, with approximately one-third of the total intake. TetraBDEs and pentaBDEs were the homologues showing the highest percentages of contribution to the sum of total PBDEs. The comparison of the current dietary intake with the suggested lowest observed adverse effect level value of 1 mg/kg/day for the most sensitive endpoints for toxic effects of PBDEs results in a safety factor over 5 orders of magnitude in relation to PBDE exposure from food.

Adult↗

Fishing in urban New Jersey: ethnicity affects information sources, perception, and compliance.

Recreational and subsistence angling are important aspects of urban culture for much of North America where people are concentrated near the coasts or major rivers. Yet there are fish and shellfish advisories for many estuaries, rivers, and lakes, and these are not always heeded. This paper examines fishing behavior, sources of information, perceptions, and compliance with fishing advisories as a function of ethnicity for people fishing in the Newark Bay Complex of the New York-New Jersey Harbor. We test the null hypothesis that there were no ethnic differences in sources of information, perceptions of the safety of fish consumption, and compliance with advisories. There were ethnic differences in consumption rates, sources of information about fishing, knowledge about the safety of the fish, awareness of fishing advisories or of the correct advisories, and knowledge about risks for increased cancer and to unborn and young children. In general, the knowledge base was much lower for Hispanics, was intermediate for blacks, and was greatest for whites. When presented with a statement about the potential risks from eating fish, there were no differences in their willingness to stop eating fish or to encourage pregnant women to stop. These results indicate a willingness to comply with advisories regardless of ethnicity, but a vast difference in the base knowledge necessary to make informed risk decisions about the safety of fish and shellfish. Although the overall median income level of the population was in the $25,000-34,999 income category, for Hispanics it was on the border between $15,000-24,999 and $25,000-34,999.

Black or African American↗

Rapid screening of fish tissue for polychlorinated dibenzo-p-dioxins and dibenzofurans.

A rapid, simple method for screening polychlorinated dibenzo-p-dioxin and dibenzofuran classes in shellfish tissue at pg g-1 wet mass concentrations is described. The method does not require a clean room facility and is based on saponification followed by extraction into hexane, clean-up using gel-permeation chromatography and sulfuric acid treatment, and measurement using capillary gas chromatography-low resolution mass spectrometry with selected ion monitoring. Selected ion monitoring using multiple ions eliminates interferences not removed by clean-up or chromatography. The detector response factor is constant for isomers within a class, e.g., tetrachlorodibenzo-p-dioxin isomers gave a mean response of 0.977 +/- 0.075 area counts fg-1, but varied significantly between classes. Thus one isomer serves as a 'standard' for all members of its class. Recoveries of added polychlorinated dibenzo-p-dioxins and dibenzofurans (20-500 pg g-1 wet mass) averaged 95.6 +/- 6.9 and 99.0 +/- 5.7%, respectively. The limits of detection (five times the noise level) are 20 pg g-1 tetrachlorodibenzo-p-dioxins and dibenzofurans, 20 pg g-1 pentachlorodibenzo-p-dioxins and dibenzofurans, 40 pg g-1 hexachlorodibenzo-p-dioxins and dibenzofurans, 40 pg g-1 heptachlorodibenzo-p-dioxins and dibenzofurans, and 100 pg g-1 octachlorodibenzo-p-dioxin and dibenzofuran. Above the limits of detection, the method gave results for polychlorinated dibenzo-p-dioxin and dibenzofuran classes in shellfish tissue comparable to those obtained by gas chromatography-high resolution mass spectrometry.

Animals↗

A case of type F botulism in southern California.

BACKGROUND: Botulism caused by type F botulinum toxin accounts for less than 0.1% of all human botulism cases and is rarely reported in the literature. CASE REPORT: A 45-year-old woman presented to an emergency department complaining of blurred vision, difficulty focusing, and dysphagia. The treating physician initially considered the possibility of paralytic shellfish poisoning due to a report of shellfish ingestion, which was later determined to be frozen shrimp and a can of tuna, but no gastroenteritis or paresthesias were present. During the emergency department observation, the patient developed respiratory distress with hypercapnea and required intubation and mechanical ventilation. Within hours, ptosis, mydriasis, and weakness in the arms and legs developed. Bivalent (A, B) botulinum antitoxin was administered approximately 24 h from the onset of initial symptoms, but over the next two days complete paralysis progressed to the upper and lower extremities. Shortly thereafter a stool toxin assay demonstrated the presence of type F botulinum toxin. The patient subsequently received an experimental heptavalent botulinum antitoxin on hospital day 7 but paralysis was already complete. Her three-week hospital course was complicated by nosocomial pneumonia and a urinary tract infection, but she gradually improved and was discharged to a rehabilitation facility. Anaerobic cultures and toxin assays have yet to elucidate the source of exposure. CONCLUSION: We report a rare case of type F botulism believed to be foodborne in etiology. Administration of bivalent botulinum antitoxin did not halt progression of paralysis.

Animals↗

Sensitization to king scallop (Pectin maximus) and queen scallop (Chlamys opercularis) proteins.

OBJECTIVE: To report a case of occupational asthma and urticaria due to the queen scallop (Chlamys opercularis) and king scallop (Pectin maximus). BACKGROUND: A 40-year-old female worked in a shellfish-processing plant, handling king and queen scallops for 5 years. At the time of investigation, she described a 2-year history of work-related respiratory symptoms. METHODS: Serial peak expiratory flow rate readings were recorded and an OASYS study completed. A workplace visit was undertaken and specific immunoglobulin (IgE) radioallergosorbent (RAST) testing of scallop extracts was performed. RESULTS: The OASYS study was consistent with occupational asthma. RAST testing demonstrated evidence of specific sensitization (IgE) to queen and king scallop. There was also some cross-reactivity observed with other shellfish (prawns and crabs). CONCLUSION: Workers exposed to aerosols from scallop species are at risk of occupational asthma and require effective respiratory health surveillance.

Adult↗

The functional characteristics conserved in tropomyosins.

1. Tropomyosin, one of the regulatory proteins in muscle contraction, was prepared from chickens, rabbits, frogs, shrimps, and shellfish, and conserved characteristics were studied using an enzymological technique. 2. All tropomyosins tested, irrespective of their sources, were found to have the ability to mediate the inhibitory activity of rabbit troponin toward rabbit Mg2+-activated actomyosin ATPase (Mg2+-ATPase) activity in the absence of Ca2+ ions. 3. The effect of tropomyosin on the Mg2+-ATPase activity in the presence of Ca2+ ions varied, depending on the source, and this variation appeared to reflect the evolutionary course of this protein. 4. Tropomyosin from shellfish adductor muscle had the ability to bind to rabbit skeletal muscle troponin and actin. This ability is also considered to be a basic characteristic of tropomyosin which has been conserved during evolution.

Actins↗

Radiation doses to critical groups since the early 1950s due to discharges of liquid radioactive waste from Sellafield.

First, some of the early work is reviewed on exposure pathways in connection with proposed and early liquid radioactive waste discharges from Sellafield. The main historical features of these discharges, affected by relevant plant operations, are then briefly described. The important radiological exposure pathways resulting from the discharges and people's consumption and occupancy habits are considered. To place the changing scenario onto a consistent basis using present-day methodology, a reconstruction of exposures has been carried out using environmental monitoring data and models. The three major pathways are examined of Porphyra/laverbread consumption in South Wales, fish and shellfish consumption near Sellafield, and external exposure over local and more distant sediments. The results show that over the period 1952 to about 1970 the laverbread pathway was probably critical, taking a cautious approach. Effective dose rates fluctuated at around 1 mSv y(-1) from about 1956 to 1971. From about 1970 to 1985, the fish and shellfish pathway was likely to have been critical, with effective dose rates peaking at about 2 mSv y(-1) in 1975-1976. External exposure was likely to have been of lesser importance than the other two pathways until about 1985, when with the retention of previously-released radiocesium on sediments it has become dominant. This phenomenon applies particularly further afield where radiocesium concentrations have been slower to decline; in the Ribble estuary, houseboat dwellers have been the critical group from about 1985. Effective doses have been at about 0.3 mSv y(-1) and declining; they are due to the effects of radiocesium discharges in earlier years. Dose rates have remained within contemporary ICRP dose limits.

Actinoid Series Elements↗

The effects of E-beam irradiation and microwave energy on Eastern Oysters (Crassostrea virginica) experimentally infected with Cryptosporidium parvum.

Shellfish have been identified as a potential source of Cryptosporidium infection for humans. The inactivation of C. parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The present study examined the efficacy of two alternative food-processing treatments, e-beam irradiation and microwave energy, on the viability of C. parvum oocysts in Eastern Oysters (Crassostrea virginica), which were artificially infected with the Beltsville strain of C. parvum. The effects of the treatments were evaluated by oral feeding of the processed oyster tissues to neonatal mice. Significant reductions (P<0.05) in infectivity were observed for in-shell and shucked oysters treated with e-beam irradiation at doses of 1.0, 1.5, or 2 kGy vs. untreated controls. A dose of 2 kGy completely eliminated C. parvum infectivity and did not adversely affect the visual appearance of the oysters. Oyster tissue treated with microwave exposures of 1 s (43.2 degrees C), 2 s (54.0 degrees C), and 3 s (62.5 degrees C) showed a reduction in C. parvum mouse infectivity, but the effects were not significantly different (P>0.05) from controls. Microwave energy treatments at 2 and 3 s showed extensive changes in oyster meat texture and color. Thus, because of lack of efficacy and unacceptable tissue changes, microwave treatment of oysters is not considered a viable food-processing method.

Animals↗