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Prevalence of seafood allergy in the United States determined by a random telephone survey.

BACKGROUND: Seafood allergy is potentially severe, but the prevalence of this group of food allergies in the US population has not been determined. OBJECTIVE: To estimate the prevalence of seafood (fish, shellfish) allergy in the United States. METHODS: We performed a nationwide, cross-sectional, random telephone survey by using a standardized questionnaire. Criteria were established in advance to define seafood allergy by report of convincing symptoms and physician evaluation. RESULTS: A total of 5529 households completed the survey (67.3% participation rate), representing a census of 14,948 individuals. Fish or shellfish allergy defined by established criteria was reported in 5.9% (95% CI, 5.3%-6.6%) of households and among individuals as follows: 2.3% (95% CI, 2%-2.5%) for any seafood allergy, 2% for shellfish, 0.4% for fish, and 0.2% for both types. Seafood allergy was more common in adults compared with children (2.8% vs 0.6%; P <.001) and in women compared with men (3.6% vs 2%; P <.001). Recurrent reactions were reported by 58%, dyspnea or throat tightness was reported by more than 50%, and 16% were treated with epinephrine. Despite this level of acuity, only 8.6% were prescribed self-injectable epinephrine. The rate of reactions to multiple fish among those with any fish allergy was 67%; for Crustacea the rate was 38%, and for mollusks the rate was 49%; only 14% with crustacean allergy reported a mollusk allergy. CONCLUSIONS: Physician-diagnosed and/or convincing seafood allergy is reported by 2.3% of the general population, or approximately 6.6 million Americans. Affected individuals typically report recurrent and sometimes severe reactions, indicating that seafood allergy represents a significant health concern.

Adult↗

Cloning, expression and characterisation of a single-chain Fv antibody fragment against domoic acid in Escherichia coli.

Domoic acid is a potent neuroexcitatory toxin that causes amnesic shellfish poisoning in humans through ingestion of contaminated shellfish. The variable regions of the heavy chain (V(H)) and light chain (V(L)) of an antibody specific for domoic acid were cloned from a mouse hybridoma cell line and used to construct single-chain antibody fragments (scFvs) in a variety of formats. V(H)-linker-V(L) scFvs were expressed better in Escherichia coli than the V(L)-linker-V(H) format, while use of the commonly used (Gly4Ser)3 inter-domain linker resulted in higher yields than a longer (Gly4Ser)6 linker variant. Higher soluble protein yields were achieved in E. coli TOP 10 than in E. coli XL1-Blue cells and co-production of the E. coli disulfide bond isomerase enzyme DsbC allowed higher cell densities to be attained during scFv production, leading to increased yields of recombinant protein. The purified scFv exhibited binding similar to the parent monoclonal antibody and is being used to develop an immunosensor to detect domoic acid in contaminated shellfish samples.

Animals↗

Ecological characterization of toxic phytoplankton species (Dinophysis spp., Dinophyceae) in Slovenian mariculture areas (Gulf of Trieste, Adriatic Sea) and the implications for monitoring.

Diarrhetic shellfish poisoning (DSP) events are often registered in Slovenian mariculture areas (Gulf of Trieste, Adriatic Sea) and are related to the occurrence of Dinophysis spp. The annual dynamic of this genus and succession of the most important species were studied at two shellfish farms during monitoring fieldwork in the period 1995-2003. Results indicate that the Dinophysis genus maintains a relatively stable inter-annual dynamic at both sites. The Dinophysis community is characterized by two surface maxima in June and September, while in the middle layer only the autumn peak is pronounced (peak median 92 cells l(-1)). Occasional abundance maxima of around 2000 cells l(-1) in the surface layer indicate that potential outbursts of toxic species are less predictable than their seasonal dynamic. On the basis of multivariate analysis, Dinophysis sacculus was characterized as a typical late spring-early summer species, and Dinophysis caudata and Dinophysis fortii as autumn species. Correlation analysis revealed the influence of stratified conditions only on the most abundant species, D. sacculus. Ecological characteristics of the species were combined with shellfish safety requirements towards a more effective monitoring.

Animals↗

Methods for isolation and confirmation of Vibrio vulnificus from oysters and environmental sources: a review.

The gram-negative bacterium Vibrio vulnificus is a natural inhabitant of estuarine waters and poses a significant health threat to humans who suffer from immune disorders, liver disease, or hemochromatosis (iron overload). V. vulnificus enters human hosts via wound infections or consumption of raw shellfish (primarily oysters), and infections frequently progress to septicemia and death in susceptible individuals. Prevalence in waters and shellfish is not correlated with fecal indicator organisms; therefore, species-specific detection and enumeration of V. vulnificus in the environment has become a priority for agencies that are responsible for shellfish safety. The many selective-differential media developed for isolation of Vibrio spp., and specifically for V. vulnificus detection, are reviewed here; however, none of the media developed to date combines the sensitivity to low numbers with the specificity necessary to inhibit growth of other organisms. Therefore, immunological and molecular protocols are needed for confirmation of the identity of the organism and are discussed in detail. Methods under development that hold promise for rapid, accurate, and sensitive detection and enumeration of the organism include multiplex and real-time PCR. Developing technologies that have proven useful for detection and investigation of other pathogens such as biosensors, spectroscopy and microarrays may provide the next generation of tools for investigation of the prevalence and ecology of V. vulnificus.

Animals↗

Teratogenic effects of azaspiracid-1 identified by microinjection of Japanese medaka (Oryzias latipes) embryos.

Azaspiracid-1 (AZA-1) is a newly identified phycotoxin that accumulates in commercially important bivalve molluscs harvested in several European countries and causes severe human intoxications. Molluscan shellfish are known vectors for accumulation and subsequent transfer of phycotoxins such as brevetoxin and domoic acid through various trophic levels within food webs. Finfish can also accumulate phycotoxins, both directly from toxic algae or from consumption of contaminated shellfish and smaller intoxicated fish. To evaluate the teratogenic potential of AZA-1 and its relevancy to toxin accumulation in finfish, we have utilized a microinjection technique to mimic the maternal-egg toxin transfer of an AZA-1 reference standard and a shellfish extract containing azaspiracids in an embryonic Japanese medaka (Oryzias latipes) fish model. Microinjection of purified AZA-1 caused dose-dependent effects on heart rate, developmental rate, hatching success, and viability in medaka embryos. Within 4 days of exposure to doses > or = 40 pg AZA-1/egg, substantial retardation in development was observed as reduced somatic growth and yolk absorption, and delayed onset of blood circulation and pigmentation. Embryos treated to > or =40 pg AZA-1/egg had slower heart rates (bradycardia) for the 9 days in ovo period, followed by reduced hatching success. Microinjection of a contaminated mussel (Mytilus edulis) extract containing AZAs (AZA-1, -2, and -3), okadaic acid, and dinophysistoxin-2 resulted in similar responses from the fish embryos at equivalent doses. These studies demonstrate that AZA-1 is a potent teratogen to finfish. This work will complement future investigations on AZA-1 accumulation in marine food webs and provide a basis for understanding its toxicity at different trophic levels.

Animals↗

Tissue distribution, effects of cooking and parameters affecting the extraction of azaspiracids from mussels, Mytilus edulis, prior to analysis by liquid chromatography coupled to mass spectrometry.

This study used liquid chromatography coupled to mass spectrometry to identify some parameters important in the analysis of azaspiracids. The first aspect was the distribution of azaspiracids within mussels, in particular the content in the digestive gland as compared to the remaining tissues. In our study, azaspiracids accumulated in the digestive gland, similar to other lipophilic toxins. The ratio of toxin in the digestive gland compared to the whole mussel was on average circa 5, both for a bulk sample collected in Norway in 2004 and for 28 samples from Ireland collected over 3 years (2001-2003). These results may justify the practise to only analyse the digestive gland, a step considered necessary to achieve adequate detection limits for azaspiracids both in the mouse bioassay and other analytical techniques. Steaming of mussels as a sample pre-treatment was found to be another parameter affecting the result. Azaspiracids concentrated indirectly, i.e. through the loss of water or juice from the matrix. The cooked shellfish tissues had a concentration of azaspiracids 2-fold higher than the uncooked shellfish, both for whole flesh and for digestive gland tissue. This finding is of particular importance since it may affect the maximum guidance level at which shellfish may be allowed for human consumption. Finally, parameters affecting the extraction efficiency were studied, including the nature of the extraction solvent, the sample-to-solvent ratio and replicate extraction. The largest differences were observed between different solvents and between different sample-to-solvent ratios, while the effect of replicate extraction was minimal if large sample-to-solvent ratios were used. Duplicate extraction using 100% methanol was found to be the best combination of parameters.

Animals↗

Further improvements in the application of high-performance liquid chromatography, capillary electrophoresis and capillary electrochromatography to the analysis of algal toxins in the aquatic environment.

The presence of algal toxins in the aquatic environment represents an important socioeconomic concern in many places worldwide, due to the toxicity that these compounds can induce in seafood or freshwater organisms at very low levels. Several analytical alternatives have been proposed over the last years for the control of these contaminants, which acute or chronic toxicity requires low detection levels and demands for the search of sensitive methods for their detection and determination. HPLC has been widely used for this purpose, although several alternatives such as CE or capillary electrochromatography (CEC) are being lately developed with this aim. In this work we report on the application of improved HPLC, as well as CE and CEC, for the analysis of diarrhetic shellfish poisoning toxins, amnesic shellfish poisoning (ASP) toxins and microcystins (MCs) present in different matrices such as water, shellfish or algae. Improvements in sample preparation for increasing sensitivity and selectivity are also shown. While UV and fluorimetric detection are the detection methods generally used, mass spectrometric detection was also applied for ASP toxins and MCs, especially for confirmatory purposes. From the results obtained it can be concluded that both HPLC and CE offer a good potential for a sensitive and selective determination of these algal toxins in such complex matrices. The results obtained for CEC allow also to conclude that this technique can result in a promising technique for such application.

Chromatography, High Pressure Liquid↗

New fluorimetric method of liquid chromatography for the determination of the neurotoxin domoic acid in seafood and marine phytoplankton.

Domoic acid (DA) is a neurotoxic amino acid that is responsible for the human toxic syndrome, amnesic shellfish poisoning (ASP). A new rapid, sensitive liquid chromatographic (LC) method has been developed for the determination of DA in various marine samples. DA in marine biological materials was derivatised with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and analysed using isocratic reversed-phase LC with fluorimetric detection. The calibration, based on standard DA solutions, was linear in the range 0.04-2 microg/ml (r2=0.998) and the detection limit (3:1, signal/noise) was better than 1 ng/ml. Using the certified reference material (MUS-1B), recoveries of DA from shellfish tissue were >95% (n=5). When a strong anion exchange SPE cartridge was used for sample clean-up the detection limit was 6 ng DA/g mussel tissue. Good reproducibility was achieved with RSD values ranging from 3% for 8 microg DA/g (n=5), to 5% for 0.04 microg DA/g (n=5). This new method was successfully applied to the determination of DA in naturally contaminated shellfish and in marine phytoplankton cultures of Pseudonitzschia sp.

Chromatography, Liquid↗

Improved method for the recovery of hepatitis A virus from oysters.

Hepatitis A is one of the major infectious diseases epidemiologically associated with worldwide shellfish consumption. Molecular detection using polymerase chain reaction (PCR) to detect hepatitis A virus (HAV) in contaminated shellfish can be hindered by low virus recoveries during the concentration process and by natural PCR inhibitors in shellfish. This study evaluated and modified two major steps of a processing procedure for virus concentration from oysters: acid adsorption-elution and solvent extraction. With the addition of second and third elutions, the acid adsorption-elution step doubled the recovery to 46% of HAV seeded initially. Extraction with chloroform or chloroform-butanol resulted in lower HAV detection limits by reverse transcription-PCR (RT-PCR)-oligoprobing than extraction with the fluorocarbon, Freon. These results led to the following modified procedure: HAV was acid adsorbed at pH 4.8, eluted first with 0.05 M glycine, second with 0.5 M threonine, PEG-precipitated twice, chloroform-extracted twice, RNA-extracted, and RT-PCR (single round) amplified. Using the modified procedure, HAV was detected by RT-PCR in all trials with a seeding density of > or = 1 plaque forming unit (PFU)/g of oyster, and in which the equivalent detection limit was 0.33 PFU of HAV seeded per RT-PCR reaction (corresponding to 111 PCR units). The method developed is capable of detecting low levels of HAV in oysters environmentally contaminated.

Animals↗

A semi-quantitative seafood safety risk assessment.

As part of a semi-quantitative risk assessment of 10 seafood hazard/product combinations, a risk assessment tool was used to generate a Risk Ranking. The tool is in a spreadsheet software format and provides a risk estimate, which is scaled between 0 and 100, where 0 represents no risk and 100 represents all meals containing a lethal dose of the hazard. A full description of the tool is contained in Ross and Sumner (this issue). Based on their ranking, seafoods in Australia fell into three risk categories. Hazard/product pairs with ranking < 32 included mercury poisoning (Relative Risk = 24), Clostridium botulinum in canned fish (RR = 25), or in vacuum-packed cold-smoked fish (RR = 28), parasites in sushi/sashimi (RR = 31), viruses in shellfish from uncontaminated waters, (RR = 31), enteric bacteria in imported cooked shrimp (RR = 31) and algal biotoxins from controlled waters (RR = 31). It is noted that there have been no documented cases of food-borne illness from any of the above hazard/product pairings in Australia. Those with rankings 32-48 included Vibrio parahaemolyticus in cooked prawns (RR = 37), V. cholerae in cooked prawns (RR = 37), Listeria monocytogenes in cold-smoked seafoods (RR = 39), scombrotoxicosis (RR = 40), V. vulnificus in oysters (RR = 41), ciguatera in the general Australian population (RR = 45), L. monocytogenes in susceptible (RR = 45) and extremely susceptible populations (RR = 47) and enteric bacteria in imported cooked shrimp eaten by vulnerable consumers (RR = 48). Almost all the hazard/product pairs in this category have caused the outbreaks of food poisoning in Australasia. Those hazard/product pairs with rankings >48 included ciguatera from recreational fishing in susceptible areas (RR = 60), viruses in shellfish from contaminated waters (RR = 67) and algal biotoxins from uncontrolled waters in an algal event (RR = 72). There have been significant (>100 cases) food poisoning incidents involving viruses and biotoxins in shellfish, while ciguatera poisoning is prevalent among coastal communities in Australia's warmer waters.

Australia↗

Incidence of and risk factors for hepatitis A in Italy: public health indications from a 10-year surveillance. SEIEVA Collaborating Group.

BACKGROUND/AIMS: This study aimed to evaluate the incidence of and risk factors for acute viral hepatitis A (HAV) in Italy. METHODS: Data were from a surveillance system for type-specific acute viral hepatitis (SEIEVA). To estimate the association of hepatitis A cases with the potential risk factors (Odds Ratios) and the proportion of all hepatitis A cases attributable to a given risk factor (population attributable risk), hepatitis B cases were used as controls. Independent predictors of HAV were estimated by conditional multiple logistic regression. RESULTS: During the period 1985-1994, 25553 viral hepatitis cases were reported. Of these, 6408 (25%) were due to hepatitis A (HAV). HAV incidence declined from 10/100000 in 1985 to 2/100000 in 1987. Since 1991, however, an increase in HAV has been observed. The majority of cases were 15-24 years old; the incidence was higher in males and in subjects residing in southern Italy. Only one death (0.02%) was observed. Shellfish consumption was the most frequently reported risk factor (62%). The proportion of cases reporting personal contact with an icteric case, travel to a high-medium endemic areas, and family contact with a child attending a day-care centre (household of day-care child) was 17%, 19% and 15%, respectively. The results of multivariate analysis showed that shellfish consumption (OR=2.6; 95% CI=2.4-2.9), travel to endemic areas for people residing in northern and central Italy (OR=5.4; 95% CI=4.6-6.2) and household of day-care child (OR=1.2; 95% CI=1.03-1.4), were all types of exposure independently associated with HAV. The estimates of population-attributable risk show that shellfish consumption explained as many as 42.2%, travel to high-medium endemic areas for people residing in northern and central Italy 24.2%, and household of day-care child only 1.4% of all acute hepatitis A cases in Italy. CONCLUSIONS: These findings indicate that HAV in Italy is mainly a food-borne disease. Vaccination against hepatitis A is strongly recommended for travellers to endemic areas.

Adolescent↗

A survey of arsenic species in Chinese seafood.

In the present report, thirty different types of Chinese edible seafood, including brown algae, red algae, fish, crab, shrimp, mussels, oysters, and clams, which are very popular foodstuffs in the Chinese kitchen, were examined for their total content of As as well as its different species. Total arsenic concentration in algae samples was 1.7-38.7 microg/g (dry weight), and 0.086-7.54 microg/g in fish and shellfish (wet weight), respectively. The arsenic species in seafood extracts were determined by using anion and cation exchange high performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICPMS). Arsenosugars were detected in all of the extracted algae samples (1.5-33.8 microg/g dry weight) and fish samples (0.018-0.78 microg/g wet weight). Arsenobetaine was detected in all of the extracted fish and shellfish samples (0.025-6.604 microg/g wet weight). In contrast, inorganic arsenic in fish and shellfish samples occurred at levels below 2% of the total arsenic. No inorganic arsenic was detected in the algae samples. This study provides information about the distribution pattern of arsenic species in seafood products. Since the major share of arsenic components in seafood is organic arsenic with a low toxicity, we can conclude that arsenic in seafood does not pose any risk to human health.

Arsenicals↗

Insufficient calcium and iron intakes among general female population in Japan, with special reference to inter-regional differences.

Dietary intakes of calcium (Ca) and iron (Fe) were investigated in 227 women (mostly housewives) in 12 regions in Japan in 1991-1993 by the 24-hour food duplicate method. Nine regions out of 12 had been previously studied in 1977-1982. Utilizing Standard Food Composition database, mean Ca and Fe-intakes in 1991-1993 were estimated to be 602 and 10.4 mg/day, respectively; the former was barely sufficient and the latter was below sufficiency when compared with the Recommended Daily Allowance in Japan for pre-menopausal women. Ca- and Fe-intake did not increase in the 10-year period. Further analysis after classification of the women into three groups of farmers in Okinawa, farmers in Mainland Japan and urban residents showed that Ca and Fe insufficiency was most evident among Okinawa farmers. The leading Ca sources were milk, pulse, vegetables and fish-shellfish, but consumption of milk was generally low, especially among Okinawa farmers. Pulse, vegetables and fish-shellfish were 3 major Fe sources; Okinawa farmers depended more on vegetables and less on fish-shellfish.

Calcium↗

The human pathogenic vibrios--a public health update with environmental perspectives.

Pathogenic Vibrio species are naturally-occurring bacteria in freshwater and saline aquatic environments. Counts of free-living bacteria in water are generally less than required to induce disease. Increases in number of organisms towards an infective dose can occur as water temperatures rise seasonally followed by growth and concentration of bacteria on higher animals, such as chitinous plankton, or accumulation by shellfish and seafood. Pathogenic Vibrio species must elaborate a series of virulence factors to elicit disease in humans. Activities which predispose diarrhoeal and extraintestinal infections include ingestion of seafood and shellfish and occupational or recreational exposure to natural aquatic environments, especially those above 20 degrees C. Travel to areas endemic for diseases due to pathogenic Vibrio species may be associated with infections. Host risk factors strongly associated with infections are lack of gastric acid and liver disorders. Involvement of pathogenic Vibrio species in cases of diarrhoea should be suspected especially if infection is associated with ingestion of seafood or shellfish, raw or undercooked, in the previous 72 h. Vibrio species should be suspected in any acute infection associated with wounds sustained or exposed in the marine or estuarine environment. Laboratories serving coastal areas where infection due to pathogenic Vibrio species are most likely to occur should consider routine use of TCBS agar and other detection regimens for culture of Vibrio species from faeces, blood and samples from wound and ear infections.

Animals↗

Structure determination of carboxyhomoyessotoxin, a new yessotoxin analogue isolated from adriatic mussels.

The contamination of shellfish with marine biotoxins derived from microalgae represents a serious problem for shellfish industries and public health. This study investigated the composition of diarrhetic shellfish toxins in the digestive glands of mussels from the Northern Adriatic Sea. Along with known yessotoxins, identified by comparison of their chromatographic and spectral properties with those reported in the literature, we isolated a new analogue of yessotoxin, carboxyhomoyessotoxin, whose structure was determined by mass spectrometry and (1)H NMR spectroscopy.

Animals↗

Toenail mercury and dietary fish consumption.

New England is one of three areas in the United States with the highest annual deposition of mercury, an established environmental pollutant with a variety of health effects. We measured the mercury content in toenails of 27 individuals in New Hampshire who participated as controls in a health study in 1994-95. The mean total toenail mercury concentration was 0.27 mcg/g (median 0.16; SD 0.27; range 0.04-1.15 mcg/g). The best predictor of toenail mercury levels was the mean combined fish and shellfish consumption measured using four simple questions from a validated food frequency questionnaire. Toenail total mercury content was significantly correlated with the mean average weekly consumption of finfish and shellfish (Spearman correlation coefficient 0.48, P=0.012). Multivariate models confirmed that toenail total mercury concentration was best predicted by total finfish and shellfish consumption.

Animals↗

Dietary patterns are associated with sexual maturation in Korean children.

The purpose of this study was to investigate the association between dietary patterns and sexual maturation among Korean children. A cross-sectional study was conducted in 422 boys and 365 girls aged 9-12 years living in Seoul, Korea. Three-day food records were obtained, and pubertal stages were determined with a questionnaire using Tanner stages. Body fat was measured by bioelectrical impedance analysis, and bone mass content at the right calcaneus was measured by portable dual-emission X-ray absorptiometry. Exploratory factor analysis with a Varimax rotation was applied to identify dietary patterns using twenty-four food groups. Four distinct dietary patterns--"rice and Kimchi", "shellfish and processed meat", "pizza and drinks" and "milk and cereal"--were obtained. Twenty-six per cent (24 % stage 2, 2 % stage 3) of boys had genital development, and 79 % (63 % stage 2, 16 % stage 3) of girls showed breast development. In boys, genital development was weakly positively associated with "shellfish and processed meat" dietary factor scores (odds ratio 1.65, CI 0.95, 2.89, P for trend 0.07) after adjusting for confounders. In girls, breast development was significantly positively associated with the factor score of "shellfish and processed meat" (odds ratio 1.88, CI 1.08, 3.26, P for trend 0.05). These results suggest that dietary patterns were related to body composition and sexual maturation among the Korean children. Further investigations are needed to identify components of the foods consumed in high amounts in these patterns and how they are related to sexual maturation.

Anthropometry↗

US Food and Drug Administration survey of cadmium, lead and other elements in clams and oysters.

In Fiscal Years 1985/1986, the US Food and Drug Administration conducted a survey of cadmium, lead and other elements in fresh clams and oysters collected from US coastal areas in use for shellfish production. Shellfish were analysed for cadmium and lead by using a dry ash-anodic stripping voltammetric method. Other elements (aluminium, arsenic, beryllium, calcium, chromium, cobalt, copper, iron, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, selenium, sodium, strontium, vanadium and zinc) were determined by using inductively coupled plasma-atomic emission spectrometry, direct current plasma-atomic emission spectrometry or hydride generation atomic absorption spectrometry. A total of 75 hardshell clam, 59 softshell clam, 104 Eastern oyster and 40 Pacific oyster samples were analysed for cadmium and lead. Average levels found were 0.09, 0.05, 0.51 and 1.1 mg/kg wet weight for cadmium and 0.24, 0.30, 0.11 and 0.06 mg/kg wet weight for lead in hardshell clams, softshell clams, Eastern oysters and Pacific oysters, respectively. The other 19 elements were determined in 10-104 samples of the four types of shellfish. These data provide baseline values for elements in clams and oysters harvested from US coastal waters.

Animals↗