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PubMedDate not supplied

The effect of commercial processing on the paralytic shellfish poison (PSP) content of naturally-contaminated Acanthocardia tuberculatum L.

A study was undertaken to determine if any reduction in contamination of Acanthocardia tuberculatum L. (Mediterranean cockle) by paralytic shellfish poisons (PSP) could be enhanced by operations carried out during the industrial canning process, allowing contaminated raw material to be commercially marketed in safe conditions for edible purposes. A general decrease in PSP levels was consistently observed when comparing raw materials and their corresponding final products, these dropping to acceptable levels. PSP levels were determined by mouse bioassay and a fluorometric method, and saxitoxin was determined by HPLC. The detoxifying effects averaged over 71.7% and 81.8% (mouse bioassay), 70.6% and 90.9% (fluorometric method), 77.9% and 83.5% (HPLC), for boiling and sterilizing operations respectively. The highest level detected in raw material was 800 micrograms/100 g by mouse bioassay.

Animals↗
PubMedDate not supplied

Two formats of enzyme immunoassay for the detection of saxitoxin and other paralytic shellfish poisoning toxins.

A competitive direct enzyme-linked immunofiltration assay for the detection of saxitoxin was developed, using polyclonal antibodies against saxitoxin and a saxitoxin-horseradish peroxidase conjugate. The test was performed in an eight-well plastic test device, in which antibody-coated nylon membranes were pressed tightly to an absorbent cellulose layer. Saxitoxin standard or sample extract solution, saxitoxin-conjugate, and enzyme substrate/chromogen solution were sequentially added on to the membrane. The test was evaluated visually by comparing the intensity of the resulting coloured (blue) dot with that of a negative control. The detection limits for saxitoxin in buffer solution and in shellfish tissue were 4 ng/ml and 80 ng/g respectively, with an assay time of less than 15 min. Under the conditions of the immunofiltration assay, decarbamoyl-saxitoxin, gonyautoxin 2/3, and neosaxitoxin standards (in buffer) gave a positive response at concentrations of about 10 ng/ml, 40 ng/ml, and 80 ng/ml, respectively. The relative cross-reactivity of the antibody to these PSPs was similar when determined using both direct and indirect (using a saxitoxin-bovine serum albumin conjugate) competitive enzyme immunoassays in microtitre plate format. In competitive direct microtitre plate assays, the 50% binding values found for saxitoxin, decarbamoyl-saxitoxin, gonyautoxin 2/3 and neosaxitoxin were 15 pg/ml, 47.5 pg/ml, 163.5 pg/ml, and 510 pg/ml respectively. In competitive indirect microtitre assay, the respective values were 138 pg/ml, 404 pg/ml, 1582 pg/ml, and 6982 pg/ml.

Animals↗
PubMed1986

Determination of "heavy" organotin pollution of water and shellfish by a modified hydride atomic absorption procedure.

Tin speciation in aquatic environment is very complex. To the natural SnIV and methylated compounds, human activities add mainly butylated, octylated, phenylated or even methylated derivatives. The most environmentally significant, due to their high toxicity and direct introduction in water through biocidal use, are the trisubstituted ones. Several sophisticated speciation procedures have been proposed, they are not susceptible of common use. We propose a simple and fast procedure allowing routine global distinction of "heavy" tin species that are most susceptible of exerting harmful effects. This AA method use the differences in volatility of stannanes generated by reduction with NaBH4. SnIV and the methylated species have very close response coefficients whereas "heavy" compounds respond very slightly at room temperature and are eliminated in a -40 degrees C cold trap. "Heavy" tin determination in water is thus obtained by the difference between two hydride AA experiments, one performed on the untreated sample ("light tin") and the other on a UV mineralised subsample (total tin). (The mineralisation of organotins is realised by UV irradiation-2 hours--in a quartz container--yields 95-100%.) The analysis of shellfish tissue relies also on two experiments. Total tin is measured on a mineralised sample and "light tin" is obtained on a subsample "solubilised" with an Ultra Turrax homogeneizer in a diluted HCl solution.

Animals↗
PubMed1984

Trace metal residues in shellfish from Maryland waters, 1976-1980.

Levels of seven heavy metal residues, arsenic, cadmium, chromium, copper, lead, mercury and zinc were monitored in samples of the American oyster (Crassostrea virginica), the soft shell clam (Mya arenaria), the hard shell clam (Mercinaria mercinaria) and the blue crab (Callinectes sapidus). Samples were taken from the Maryland section of the Chesapeake Bay and its tributaries over a five year period (1976-80). This study was undertaken to provide an estimate of a baseline for values of trace heavy metals. Also, the oyster, being a non-mobile filter feeder, provides information regarding the level of metal residues in its environment (water and sediment), because heavy metal uptake is related to the surrounding metal concentrations. Additionally, this type of monitoring program is essential to ensure that shellfish sold for human consumption are within safe limits established for toxic substances by the United States Food and Drug Administration. Results of the study are consistent with data from previous years and no statistically significant year-to-year trends were observed over the period of the investigation.

Animals↗
PubMed2003

Chlorination for degrading saxitoxins (paralytic shellfish poisons) in water.

Chlorination was investigated as a treatment option for degrading and thus removing saxitoxins (paralytic shellfish poisons, PSPs) produced by cyanobacteria (blue-green algae) from water. It was found to be effective with the order of ease of degradation of the saxitoxins being GTX5 (B1) approximately dcSTX > STX > GTX3 approximately C2 > C1 > GTX2. However the effectiveness of chlorine was pH dependent. Degradation as a function of pH was not linear with the degree of degradation increasing rapidly at around pH 7.5. At pH 9 > 90% removal was possible provided a residual of 0.5 mg l(-1) free chlorine was present after 30 min contact time. The more effective degradation at higher pH was unexpected as chlorine is known to be a weaker oxidant under these conditions. The more effective degradation, then, must be due to the toxins, which are ionisable molecules, being present in a form at higher pH which is more susceptible to oxidation. The feasibility of using chlorine to remove saxitoxins during water treatment will therefore depend strongly on the pH of the water being chlorinated. Degradation may be improved by pH adjustment but may not be a practical solution. Although saxitoxins were degraded in that the parent compounds were not detected by chemical analysis, there is no indication as to the nature of the degradation products. However, acute toxicity as determined by the mouse bioassay was eliminated.

Animals↗
PubMed1996

Spoilage and shelf-life extension of fresh fish and shellfish.

Fresh fish and shellfish are highly perishable products due to their biological composition. Under normal refrigerated storage conditions, the shelf life of these products is limited by enzymatic and microbiological spoilage. However, with increasing consumer demands for fresh products with extended shelf life and increasing energy costs associated with freezing and frozen storage, the fish-processing industry is actively seeking alternative methods of shelf life preservation and marketability of fresh, refrigerated fish and at the same time economizing on energy costs. Additional methods that could fulfill these objectives include chemical decontamination, low-dose irradiation, ultra-high pressure, and modified atmosphere packaging (MAP). This review focuses on the biochemical and microbiological composition of fresh fish/shellfish, the spoilage patterns in these products, factors influencing spoilage, and the combination treatments that can be used in conjunction with refrigeration to extend the shelf life and keeping quality of fresh fish/shellfish. The safety concerns of minimally processed/MAP fish, specifically with respect to the growth of Clostridium botulinum type E, is also addressed.

Animals↗
PubMed1981

Lymphatic absorption of shellfish sterols and their effects on cholesterol absorption.

Studies have been conducted on the absorbability of individual sterols from a mixture of oyster sterols when administered intragastrically to rats with indwelling catheters in the left thoracic duct. In addition, the effect of oyster sterols on cholesterol absorption has been assessed using [4-14C] cholesterol in the mixture, and comparison against absorption of cholesterol alone. The order of absorbability (percentage absorption) of individual sterols from the mixture of oyster sterols was: cholesterol greater than or equal to 26-carbon sterols greater than or equal to dehydrocholesterol greater than 24-methylene cholesterol greater than brassicasterol greater than plant sterols. The absorption of noncholesterol sterols was 8.2 +/- 0.8% of the fed dose, or less than half of that for an equivalent level of cholesterol alone. The presence of these sterols in mixtures containing cholesterol reduced lymphatic absorption of cholesterol by 25 to 40% compared to absorption of the same amount of cholesterol administered alone, or to an amount of cholesterol equal to the total oyster sterols, respectively. These studies suggest that shellfish sterols are poorly absorbed, and, like plant sterols, effectively reduce dietary and/or endogenous cholesterol absorption from the intestine.

Absorption↗
PubMed1989

Recurrent epidemic hepatitis A associated with consumption of raw shellfish, probably controlled through public health measures.

Between April 1984 and January 1985, in the Italian seaport of Livorno, the annual incidence of serologically confirmed acute hepatitis A doubled to 46 per 100,000 population. The exposure histories of each of 75 jaundiced subjects with serologically confirmed hepatitis A were compared with up to four, randomly chosen-, age-, sex-, and neighborhood-matched controls. Illness was strongly associated with consumption of raw mussels and clams within six weeks of onset of illness. When the two thirds of the subjects who had been exposed were classified according to the frequency with which they had recently consumed any type of raw shellfish, there was a clear dose-response relation. In February 1985, comprehensive control measures were introduced and the annual incidence of hepatitis A fell to 2.3 per 100,000 population, a 10-fold decrease from the preepidemic period.

Adolescent↗
PubMed1982

The identification of critical groups and its application to fish and shellfish consumers in the coastal area of the North-East Irish Sea.

Control of radioactive waste disposal to the environment, based on the recommendations of the International Commission on Radiological Protection (ICRP), necessitates an identification of the critical group of members of the public exposed from a given practice. Criteria for identification of critical groups based mainly on ICRP recommendations are discussed. Methods existing up to 1974 are briefly reviewed in the light of these criteria. Two more recent methods are described and compared on the basis of previous data; these methods, which are complementary, are based on homogeneity considerations and satisfy the criteria generally rather better than do the earlier methods. The application of the newer techniques to recent surveys of fish and shellfish consumption in the coastal area of the North-East Irish Sea is described, in relation to discharges from the Windscale reprocessing plant. The results of this survey are presented, and estimates are given of the effective dose equivalent to members of these critical groups in recent years, to show the effect of liquid discharges from Windscale in terms of public radiation exposure.

Animals↗
PubMed1994

Analytical procedure for use of conductance measurement to estimate Escherichia coli in shellfish.

Assays were performed with a Malthus AT Microbiological Analyzer to define an analytical procedure to estimate Escherichia coli counts in live bivalve shellfish by conductance measurement. The growth conditions used (Malthus Coliform Broth at 44 degrees C) were selective for E. coli, and interference was noted only when Klebsiella pneumoniae were at least 100 times as numerous as E. coli. Different sample preparation procedures and seeding conditions were tested to obtain good quality conductance curves. The best results were observed when: (a) meat and shell liquor were diluted 1:3 with tryptone salt water and homogenized in a Waring blender for 1 min at 15,000 rev min-1; and (b) the inoculum was taken from the liquid phase of the homogenate 20 min after blending and mixed immediately with the culture medium. Detection parameter threshold values were adjusted (first difference 1.5 microS for the baseline and 3.5 microS for detection, second difference 0.2 microS) to improve detection time reliability. The repeatability of conductance measurements was very good (S.D. as % response mean ranged from 1.9 to 3.3) with the protocol used.

Animals↗
PubMed2001

[Detection of TDH-producing Vibrio parahaemolyticus O3:K6 from naturally contaminated shellfish using an immunomagnetic separation method and chromogenic agar medium].

We attempted to isolate TDH-producing Vibrio parahaemolyticus O3:K6 from shellfish. Asari samples were incubated with TSB supplemented with 2% (w/v) NaCl for 6 h, and then the 6-h cultures were incubated with salt polymyxin broth for 18 h. After the two-step enrichment, a 1 ml portion of the culture was treated with magnetic beads coated with K6 antibody for immunoconcentration of V. parahaemolyticus O3:K6. The immunoconcentrated and untreated cultures were plated onto a chromogenic agar and TCBS agar media for isolation of V. parahaemolyticus. TDH-producing V. parahaemolyticus O3:K6 was isolated from 3 out of 66 lots (4.5%) of naturally contaminated Asari. Six of 4,265 colonies suspected as V. parahaemolyticus (0.14%) were TDH-producing V. parahaemolyticus O3:K6.

Agar↗
PubMed1981

Transformation of paralytic shellfish toxins as demonstrated in scallop homogenates.

Toxins in shellfish, which are responsible for paralytic poisonings, undergo reductive transformation when incubated with the homogenate of various portions of the scallop, Placopecten magellanicus. The transformation includes the reductive elimination of O-sulfate groups, a change that is most evident in the locomotor tissue homogenates. The commercially important adductor muscles can also inactivate the toxins.

Animals↗
PubMed1976

Antibiotic resistance among coliform and fecal coliform bacteria isolated from sewage, seawater, and marine shellfish.

Seawater and shellfish samples collected in the vicinity of a marine sewage outfall were examined for the incidence of antibiotic resistance among coliform and fecal coliform bacteria over a 2-year period. Seventy percent or more of these two groups of bacteria from both sources were resistant to one or more antibiotics. Forty-five percent of the isolates resistant to streptomycin or tetracycline were capable of transferring all or part of their resistance pattern to an antibiotic-susceptible strain of Escherichia coli K-12.

Anti-Bacterial Agents↗
PubMed1976

Microbiological quality of frozen breaded fish and shellfish products.

A survey was made of the microbiological quality of seven frozen, breaded, precooked fish and shellfish products and of frozen, breaded, uncooked shrimp at the retail level. Geometric mean aerobic plate counts per gram (and number of units examined) were as follows: fish sticks, 8,300 (1,539); fish cakes, 5,600 (1,378); crab cakes, 4,900 (1,226); scallops, 1,700 (1,392); clams, 450 (1,384); haddock, 15,000 (1,306); fish in fish and chips dinner, 7,200 (1,485); and uncooked shrimp, 220,000 (1,462). Geometric mean coliform, Escherichia coli, and Staphylococcus aureus counts for all eight products ranged from 1 to 10/g.

Cell Count↗
PubMed1992

Development of monoclonal antibodies that identify Vibrio species commonly isolated from infections of humans, fish, and shellfish.

Monoclonal antibodies (MAbs) against Vibrio species that infect humans, fish, and shellfish were developed for application in rapid identifications. The pathogens included Vibrio alginolyticus, V. anguillarum, V. carchariae, V. cholerae, V. damsela, V. furnissii, V. harveyi, V. ordalii, V. parahaemolyticus, and V. vulnificus. Three types of MAbs were selected. The first important group included MAbs that reacted with only a single species. A second group comprised a number of MAbs that reacted with two, taxonomically closely related Vibrio species. For example, of 22 MAbs raised against V. alginolyticus, 6 recognized a 52-kDa flagellar H antigen common to both V. alginolyticus and V. parahaemolyticus; V. anguillarum and V. ordalii also shared antigens. A third group included three genus-specific MAbs that reacted with almost all Vibrio species but did not react with other members of the family Vibrionaceae (e.g., members of the Aeromonas, Photobacterium, and Plesiomonas genera) or a wide range of gram-negative bacteria representing many genera. This last group indicated the possible existence of an antigenic determinant common to Vibrio species. Two of these three genus-specific MAbs reacted with heat-stable antigenic determinants of Vibrio species as well as lipopolysaccharide extracted from Vibrio species. The use of the MAbs in blind tests and diagnosis of clinical isolates indicated that three different types of bacteria, viz., live, formalin-fixed, and sodium azide-killed bacteria, were detected consistently. Overall, it was found that the genus-specific MAbs were very useful for rapidly identifying vibrios in the screening of acute infections, while the species-specific MAbs and others were useful for completing the diagnosis.

Animals↗
PubMed1994

In situ detection of hepatitis A virus in cell cultures and shellfish tissues.

An in situ transcription method was developed to detect hepatitis A virus RNA in both cell cultures and shellfish tissues. Radiolabeled cDNA copies were synthesized in situ by reverse transcriptase-directed transcription after annealing with a specific primer to the viral RNA. Both tritium (3H) and 35S were useful in the in situ transcription reaction, but the use of 3H resulted in a lower background and finer detail in the localization of viral particles. Application of the method to different organs of oysters which had bioaccumulated hepatitis A virus allowed the first in situ localization of the virus, specifically in stomach and hepatopancreatic tissues.

Animals↗
PubMed1995

Sequence of a cloned pR72H fragment and its use for detection of Vibrio parahaemolyticus in shellfish with the PCR.

The nucleotide sequence of pR72H cloned from Vibrio parahaemolyticus 93 was determined. We examined all V. parahaemolyticus gene sequences published in the GenBank-EMBL databases for homology and found that no other DNA sequence of V. parahaemolyticus was highly homologous to the sequence reported in this study. A pair of primers, VP33-VP32, derived from a pR72H fragment were selected to detect V. parahaemolyticus. The sensitivity of PCR detection for a pure culture of V. parahaemolyticus was 10 cells from crude bacterial lysates. Furthermore, a detection level of 2.6 fg, equivalent to 1 cell, was obtained by using purified chromosomal DNA as the template. The expected PCR products were obtained from all V. parahaemolyticus strains tested (n = 124), while no PCR amplicons were found in other vibrios or related genera (n = 50). High levels (10(6) to 10(10) CFU/ml) of Escherichia coli cells did not affect the PCR assay sensitivity. The presence of 10(8) V. parahaemolyticus cells or 10(9) E. coli cells in the PCR mixtures completely inhibited the PCR. When oyster samples were inoculated with V. parahaemolyticus 93 and cultured in tryptic soy broth containing 3% NaCl for 3 h at 35 degrees C, an initial sample inoculum level of 9.3 CFU/g was detected in a PCR assay with crude bacterial lysates. The PCR assay with enrichment culturing in salt polymyxin broth was compared with the conventional method for naturally contaminated shellfish and fish samples. We conclude that this PCR assay with enrichment culturing is a good alternative method for the detection of V. parahaemolyticus.

Animals↗