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A new component of paralytic shellfish poison was isolated from a Bangladeshi freshwater puffer Tetraodon cutcutia. Its structure was deduced to be carbamoyl-N-methylsaxitoxin based on electrospray ionization mass spectrometry, [1H] NMR, and conversion experiments.
The cross-reactivity of the 6/50 monoclonal anti-okadaic acid antibody (mAb) to the recently discovered diarrhetic shellfish poisoning (DSP) metabolites dinophysistoxin-4 (DTX-4), dinophysistoxin-5 (DTX-5), and an okadaic acid (OA) diol ester was determined using a competitive indirect enzyme-linked immunosorbent assay (ELISA). The reactivity of the antibody to these molecules was compared to that with OA; the 6/50 mAb recognized all of these DSP compounds with equal sensitivity within the working range of the antibody (10-100 nM for OA). This confirms the ability of the antibody to detect all DSP compounds when used in analyses including ELISA and immunocytochemistry.
A number of algal toxins were tested for the ability to induce apoptosis (regulated cell death) in primary hepatocytes from salmon and rat. The tested toxins included the liver targeting substances microcystin-LR and nodularin, substances associated with the diarrhetic shellfish poison complex (okadaic acid, dinophysistoxin-1 and pectenotoxin-1) and calyculin A. All toxins induced apoptosis in both salmon and rat hepatocytes in less than 2 h. The apoptotic changes were evident both by electron and light microscopy and were counteracted by the caspase inhibitor ZVAD-fmk and by the Ca2+/calmodulin dependent kinase II inhibitor KN-93. The salmon hepatocytes were 10-20-fold more sensitive to okadaic acid and dinophysistoxin-1 (EC50=20 nM) than rat hepatocytes and other mammalian cell lines tested. An assay was devised using hepatocyte apoptosis as parameter for detection of algal toxins. This assay was at least as sensitive as HPLC determination for okadaic acid in mussel extracts. It also detected algal toxins which do not inhibit protein phosphatases, like pectenotoxin-1. Subapoptotic concentrations of the toxins inhibited hepatocyte aggregation. Using this parameter, less than 200 pg okadaic acid could be detected. In conclusion, salmon hepatocytes in suspension culture provide a rapid and sensitive system for detection of a broad range of apoptogenic toxins.
Animals may acquire toxicity by absorbing toxic compounds from their food, e.g. from plants or other animals. Sequestration and accumulation of toxins may provide protection from predators, which learn to avoid this prey because of unpleasant experiences such as bitter taste. This is a common phenomenon in marine as well as in terrestrial ecosystems. Moreover, toxins may enter food chains where they accumulate reaching high, often lethal concentrations. Palytoxin which had been primarily detected in marine zoanthids (Palythoa sp.), occurs also in a wide range of other animals, e.g. in sponges, corals, shellfish, polychaetes and crustaceans, but also in fish, which feed on crustaceans and zoanthids as well. These animals exhibit a high resistance to the toxin's action. The mechanisms which protect the Na+, K+-ATPase of their cell membranes, the primary target of palytoxin, is unknown. Sequestration of the toxin by other animals may cause health problems due to food poisoning.
The diarrhetic shellfish toxin composition in the hepatopancreas of mussels from the northern Adriatic sea was investigated. The major toxins were shown to be yessotoxin (YTX), homoyessotoxin (homoYTX) and 45-hydroxyyessotoxin (45-OHYTX), identified by comparison of their chromatographic and spectral properties with those reported in the literature.
Mediterranean strains of Prorocentrum minimum do not appear to have the same toxic component as Japanese strains since they showed no cytotoxicity for hepatocytes in culture. However, their toxic components, which appear to block calcium channels, were detectable by the immobilisation test on Diptera larvae. A bio-accumulation experiment in the laboratory showed that the toxins could accumulate in nearly equivalent amounts in the hepatopancreas and meat of cultured mussels. The same toxicity was found in natural samples collected in a period of bloom of P. minimum. These results suggest that P. minimum could be responsible for shellfish toxicity in the natural environment and thus present a risk for human health.
Okadaic acid was isolated from a strain of Prorocentrum arenarium Faust (Prorocentrales, Dinophyceae) collected from Europa Island (40 degrees 22'E, 22 degrees 20'S, SW Indian Ocean). The presence of okadaic acid in the algal extract was suspected after cytotoxicity and phosphatase 2A inhibition testing. It was confirmed by ADAM derivatization, immunoaffinity extraction and liquid chromatography with fluorimetric detection analysis as well as by liquid microchromatography with mass spectrometric detection. Results indicate that the P. arenarium strain was toxinogenic and could be potentially involved in the toxin production associated with the human diseases, diarrhetic shellfish poisoning and possibly ciguatera fish poisoning in the SW Indian Ocean area.
Total diet study (TDS) samples of 14 food groups from 16 locations in Japan, collected in 1999 and 2000, were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) to estimate the update of daily intake of these contaminants from food. The mean daily intake of toxic equivalency (TEQ) for an adult weighing 50 kg, calculated at non-detected isomer concentrations equal to zero (ND = 0), was estimated to be 2.25 pg TEQ/kg b.w./day. When non-detected isomer concentrations are assumed to be equal to half of the limits of detection (ND = 1/2 LOD), the mean daily intake was estimated to be 3.22 pg TEQ/kg b.w./day. These values were below the tolerable daily intake (TDI) of 4 pg TEQ/kg b.w. for PCDD/Fs and dioxin-like PCBs set in Japan. In both the estimates, the mean daily intakes were highest from fish and shellfish (76.9% at ND = 0 and 53.9% at ND = 1/2 LOD of the total TEQs), followed by those from meat and eggs (15.5% at ND = 0 and 11.7% at ND = 1/2 LOD of the total TEQs). Congener specific data revealed that these total TEQ levels were dominated by 1,2,3,7,8-PeCDD, 2,3,4,7,8-PeCDF and 3,3',4,4',5-PeCB in each case (71.7% at ND = 0 and 63.1% at ND = 1/2 LOD of the total TEQs). The dioxin-like PCBs (non-ortho and mono-ortho PCBs) accounted for about 50% of these total TEQs. These data will be very useful in the risk assessment of PCDD/Fs and dioxin-like PCBs from food in Japan.
Concentrations and accumulation profiles of PCDDs/DFs and coplanar-PCBs (co-PCBs) in aquatic biota (e.g., plankton, shellfish, benthic invertebrate, and fish) and sediment from Tokyo Bay were examined to elucidate the relationship between bioaccumulation and trophic level in the food web as determined by the stable nitrogen isotope analysis. Bioaccumulation patterns of PCDDs/DFs and co-PCBs varied greatly among congeners. Accumulation patterns of PCDDs/DFs and co-PCBs are not solely explained by their physicochemical properties. Biota-sediment accumulation factors (BSAFs) for co-PCBs in biota from Tokyo Bay were significantly greater than those of PCDDs/DFs. Furthermore, the slopes of the plots of delta15N and BSAF values and water solubility of 2,3,7,8-substituted PCDDs/DFs and co-PCBs were highly correlated. The results of our study would provide the valuable information to understand the accumulation properties of PCDDs/DFs and co-PCBs that can be used as a scientific basis to determine the sediment quality criteria of PCDDs/DFs and co-PCBs.
In the Pearl River Delta of China, the rapidly developing industrial and agricultural activities, municipal development and use of chemicals caused serious environmental problems. This report summarizes the published scientific data on POPs in the environment of the Pearl River Delta, including the levels of POPs in the air, water, soil, river and estuarine sediments, the marine organisms like fish and shellfish in this region. The data preliminarily reveal the state of contamination in this region and give insight into the fate of POPs in this sub-tropical area. However, most research in this area is limited to a few kinds of POP compounds.
From 1998 to 2000, the dioxin levels were measured in the milk of 299 mothers who lived in Saitama prefecture, Japan. Factors that influenced the dioxin levels were investigated based on a questionnaire given to the milk donors at that time. It was found that the dioxin levels in the milk of the mothers who smoked were lower than those of non-smokers, and the dioxin levels were generally low in the mothers who were heavy smokers. The average dioxin levels in the milk of mothers who had been breast-fed were higher than those given formula, and there was a significant difference in the dioxin congeners. It was also found that the dioxin levels in milk of the women who regularly consumed fish and shellfish were generally higher. There was a strong correlation between summation operator TEQ and PCB126. The data suggested that PCB126 could be a useful indicator for the simplified analysis of dioxin in human milk.
The average person in Hong Kong consumes fish or shellfish four or more times a week averaging about 60 Kg of fish per year. Even though the mean mercury level in store-bought Hong Kong fish was only 0.12 mg/kg, corroborating evidence is presented from numerous studies to support the view that mercury bioaccumulates. By the time a typical Hong Kong male reaches 30 years of age he will have accumulated approximately 4 mg/kg mercury in his hair. By age 60, his hair mercury levels will have increased to about 7.5 mg/kg. Hair is a useful indicator of mercury exposure. In the U.S. over a million hair samples have been examined for mercury (mean, 1.5 mg/kg). The mean hair mercury concentration for over 200 Hong Kong residents was 3.3 mg/kg which is more than double the U.S. mean (well over one standard deviation above the U.S. mean). Two lines of evidence support the hypothesis that fish is the major source of methyl mercury in the diet of Hong Kong residents. 1. Individuals consuming 4 or more meals of fish per week had a hair mercury of 4.07 mg/kg dry weight of hair while those consuming fish less frequently had significantly lower levels (2.56 mg/kg). 2. Hong Kong residents that consume no fish had only 0.38 mg/kg hair mercury. The World Health Organization has adopted the U.S. EPA levels for mercury and recommends that food with mercury concentrations of 0.5 mg/kg or more should not be sold for human consumption. Data presented in this paper are consistent with the notion that adoption of a 0.3 mg/kg mercury guideline would benefit residents in countries where rates of fish consumption are significantly higher than in the U.S. Japan, for example, has already adopted a 0.3 mg/kg mercury guideline. In Hong Kong there is a significant correlation (p < 0.05) between male subfertility and the level of mercury in the hair of males between the ages of 25 and 75. Our study of individuals who have been eating mercury contaminated fish steadily for many years suggests that a daily mercury intake of only 0.3 to 0.7 mg/kg body weight may be sufficient to inhibit spermatogenesis in some Hong Kong males. Male subfertility has been correlated with both elevated mercury and the presence of various organochlorines in the diet. To determine whether fish sold in Hong Kong with elevated levels of mercury also had elevated levels of organochlorines we analyzed fish for both mercury and organochlorine content of their dorsal muscle tissue. Because analysis of fish tissue for lipids and a wide range of organochlorines is both very time consuming and expensive, only 15 different species of fish were tested. Organochlorine concentrations were low and there was no correlation between mercury and organochlorine in the 15 fish tested for both organochlorines and mercury. As a result of these tests we concluded that mercury could not be ruled out as the principal causal factor associated with the lack of fertility in Hong Kong males.
Kastela Bay, located on the central part of east Adriatic coast, is heavily contaminated by mercury. The main source of contamination was a chlor-alkali plant, which was operating for 40 years since 1949. Previous studies showed that mercury concentrated in the sediment close to the plant is being dispersed throughout the bay by remobilization processes. Subsequent remobilization of mercury buried in the sediment may be harmful to various marine organisms. In order to assess the availability of remobilised mercury to marine organisms and to assess the health risk of consumption of shellfish species from the bay, a monitoring programme was initiated in September 1997. One thousand six hundred specimens of mussels (Mytilus galloprotincialis) of the same size and age, collected in a very clean area, were transferred to four stations in the bay 1.5 m above the sea bottom. Every month during the summer period and every second month in the winter period, 50 mussels were taken from each station for the analysis of mercury and monomethylmercury (MeHg) content. The results obtained during 1 year of biomonitoring suggest that the equilibrium concentration in transplanted mussels was established in a relative short period of time. The digestive gland is preferential organ for the accumulation of total mercury. Seasonal variation of mercury content both in the whole soft tissue, and organs (gills and digestive gland) was observed at all stations. However, methyl mercury is more concentrated in soft tissues than in digestive gland and gills, despite the fact that it is absorbed through these organs. The mercury levels in tissues were below accepted limits for human consumption. Obtained results showed that the implantation of mussels in an area containing sediment contaminated by mercury may be a good monitoring tool for the assessment of the availability of remobilised mercury to marine organisms.
The consumption of contaminated seafood has been reported as an important route of human exposure to metals in Taiwan. We consider the concentrations of TBT, Cu, Zn, Cd, As, and the consumption of oysters of Taiwanese to be the important information related to public health in Taiwan. Therefore, the aim of this study was to evaluate the public health risks associated with TBT, Cu, Zn, Cd and As from shellfish for the general population and fishermen of Taiwan. In general, TBT concentrations in various oysters ranging from 0.32 to 1.51 microg/g dry wt. varied with sampling locations. The highest TBT, Cu, and Zn geometric mean (GM) concentrations in oysters of 1.51, 1180 and 1567 microg/g dry wt. were obtained from the Hsiangshan coastal area. The values of oyster consumption for fishermen were 94.1 and 250 g/day for typically and maximally exposed individuals, respectively. In particular, the highest intake (250 g/day) from fishermen was almost two times greater than that of the general population (139 g/day). The THQ (target hazard quotient) values of Hsiangshan's fishermen are 3.87 and 20.50 for TBT and Cu for maximally exposed individuals are higher than other oyster culture areas. It is interesting that those consuming oysters from Hsiangshan, Lukang, Taishi caused abnormally high THQs of TBT and other metals (100% over 1.0), and TBT was attributed to only 3-21% of the total THQs in different fishermen of Taiwan. Our results suggest that current environmental levels of TBT and other metals are associated with a significant potential threat to human health for fishermen resident in coastal areas of Taiwan.
The focus of the present study was on the relationship between Hong Kong male subfertility and fish consumption. Mercury concentrations found in the hair of 159 Hong Kong males aged 25-72 (mean age = 37 years) was positively correlated with age and was significantly higher in Hong Kong subjects than in European and Finnish subjects (1.2 and 2.1 ppm, respectively). Mercury in the hair of 117 subfertile Hong Kong males (4.5 ppm, P < 0.05) was significantly higher than mercury levels found in hair collected from 42 fertile Hong Kong males (3.9 ppm). Subfertile males had approx. 40% more mercury in their hair than fertile males of similar age. Although there were only 35 female subjects, they had significantly lower levels of hair mercury than males in similar age groups. Overall, males had mercury levels that were 60% higher than females. Hair samples collected from 16 vegetarians living in Hong Kong (vegans that had consumed no fish, shellfish or meat for at least the last 5 years) had very low levels of mercury. Their mean hair mercury concentration was only 0.38 ppm.
BACKGROUND: Although cross-reactivity between mollusks and other crustaceans in shrimp-sensitive subjects has been reported, the mechanism of this allergenic cross-reactivity has not been studied in detail. OBJECTIVE: To investigate this cross-reactivity in vitro, we have taken advantage of a complementary DNA that expresses tropomyosin, the immunodominant shrimp allergen. METHODS: Serum IgE from nine patients with known anaphylaxis to shrimp and five normal volunteers were analyzed by immunoblotting against 13 distinct crustaceans and mollusks. As additional antigens, muscle preparations were isolated from grasshopper, cockroach, fruit fly, chicken, and mouse. RESULTS: Sera from all patients, but not control subjects, reacted specifically with a 38 kd protein in all crustaceans and mollusks studied. In addition, 8 of 9 sera from patients, but from none of the normal control subjects, recognized proteins of various other molecular weights among the mollusk extracts studied. The 38 kd protein was identified as tropomyosin and was shown to share immunodominant epitopes among all species of crustaceans and mollusks tested by specific absorption studies. Moreover, sera from all nine subjects with shrimp allergy demonstrated IgE reactivities against grasshopper, cockroach, and fruit fly but not chicken or murine muscle. CONCLUSION: The allergic epitopes on tropomyosin are conserved among invertebrates including not only shellfish but also insects. This latter observation suggests that persons sensitive to shrimp should undergo further study for potential cross-reactive inhalant or ingested insect sensitivity.
Up to 8% of children less than 3 years of age and approximately 2% of the adult population experience food-induced allergic disorders. A limited number of foods are responsible for the vast majority of food-induced allergic reactions: milk, egg, peanuts, fish, and tree nuts in children and peanuts, tree nuts, fish, and shellfish in adults. Food-induced allergic reactions are responsible for a variety of symptoms involving the skin, gastrointestinal tract, and respiratory tract and may be caused by IgE-mediated and non-IgE-mediated mechanisms. In part 1 of this series, immunopathogenic mechanisms and clinical disorders of food allergy are described.
Harmful algal blooms (HAB) may impose a serious threat to aquatic lives and human health. We determined the effects of a toxic dinoflagellate Alexandrium tamarense (clone ATCIO1, isolated from Hong Kong coastal waters) on the energy budget, quantified as scope for growth (SFG), and the growth rate of the manila clam Ruditapes philippinarum and the green-lipped mussel Perna viridis. To quantify the SFG, clams and mussels were dosed with different amounts of toxic A. tamarense for 6 days, resulting in different toxin burdens in the tissues. Clearance rate, absorption efficiency, and respiration rate were subsequently measured in these toxin containing bivalves. Clearance rate significantly declined at the highest toxin burden in the clams only, while there was no significant difference in the clearance rate among different treatments for the mussels. The respiration rate of either bivalve was not significantly affected by toxin accumulation in the tissues. In contrast, the absorption efficiency of both clams and mussels decreased, in a concentration-dependent manner for mussels but not for clams, when the tissue accumulated the toxins. With an increase in paralytic shellfish poisoning (PSP) toxin burden, SFG in both clams and mussels was significantly reduced, primarily because of the decrease of absorption efficiency. The growth rate of juvenile clam R. philippinarum, measured as an increment in tissue dry weight over a 15 d exposure period, was significantly lower during their feeding on toxic dinoflagellate than the growth rate of clams feeding on the diatom Thalassiosira pseudonana. The juvenile mussel P. viridis, however, exhibited similar growth rates after feeding on the toxic dinoflagellates and the nontoxic diatom. This study showed that SFG can provide a sensitive and integrated measure of the effect of HAB on the physiology of bivalves. Clam R. philippinarum may be more sensitive, in terms of their energy budget, to PSP toxin accumulation than the mussel P. viridis.