We evaluated the quality of seawater and ribbed mussels (Gukensia demissa) at six sites along the West Coast of Assateague Island National Seashore (ASIS), a barrier island popular with tourists and fishermen. Parameters evaluated were summertime temperature, pH, salinity, dissolved oxygen, total phosphorus, total ammonia nitrogen, and nitrite levels for seawater and total heterotrophic plate counts and total Vibrionaceae levels for the ribbed mussels. Approximately 150 feral horses (Equus caballus) are located on ASIS and, combined with agricultural runoff from animals and croplands, local wildlife, and anthropogenic inputs, contribute to nutrient loads affecting water and shellfish quality. The average monthly dissolved oxygen for June was 2.65 mg L(-1), below the minimum acceptable threshold of 3.0 mg L(-1). Along Chincoteague Bay, total phosphorus generally exceeded the maximum level of 0.037 mg L(-1), as set by the Maryland Coastal Bays Program management objective for seagrasses, with a high of 1.92 mg L(-1) in June, some 50-fold higher than the recommended threshold. Total ammonia nitrogen approached levels harmful to fish, with a maximum recorded value of 0.093 mg L(-1). Levels of total heterotrophic bacteria spiked to 9.5 x 10(6) cells g(-1) of mussel tissue in August in Sinepuxent Bay, leading to mussels which exceeded acceptable standards for edible bivalves by 19-fold. An average of 76% of the bacterial isolates were in the Vibrionaceae family. Together, these data suggest poor stewardship of our coastal environment and the need for new intervention strategies to reduce chemical and biological contamination of our marine resources. PMID:19132436
Lambert, Mary S; Ozbay, Gulnihal; Richards, Gary P
Although mussel beds are common in many intertidal habitats, the ecological significance of the aggregated distribution of mussels has not been examined. The ribbed mussel, Geukensia demissa, is found in dense aggregations on the seaward margin of many salt marshes in New England. Here, we examine the population structure of G. demissa in a New England salt marsh and investigate
Mark D. Bertness; Edwin Grosholz
The Atlantic ribbed musselGeukensia (Modiolus)demissa attaches itself to the roots of cord grass and other hard objects in tidal salt marshes by spinning adhesive byssal threads. The precursor of a protein apparently present in the adhesive plaques of the threads was isolated in quantity from the foot of the mussel. The protein has an apparent molecular weight of 130000, a
J. Herbert Waite; Douglas C. Hansen; Kathleen T. Little
The Atlantic ribbed mussel Geukensia (Modiolus) demissa attaches itself to the roots of cord grass and other hard objects in tidal salt marshes by spinning adhesive byssal threads. The precursor of a protein apparently present in the adhesive plaques of the threads was isolated in quantity from the foot of the mussel. The protein has an apparent molecular weight of 130,000, a pI of 8.1, and contains a high proportion of Gly, Glu/Gln, Lys and 3,4-dihydroxyphenyl-L-alanine (DOPA). Sequence of tryptic peptides suggests a pattern of repeated motifs, such as: Gly--DOPA--Lys, and X--Gly--DOPA--Y--Z--Gly--DOPA/Tyr--Lys, where X is Thr or Ala in octapeptides and Gln--Thr in nonapeptides. Y is variable, but more often than not hydrophobic; and Z is frequently Pro or 4-trans-hydroxyproline (Hyp). The presence of Pro--Gly and Hyp--Gly sequences of delta-hydroxylysine in the protein is reminiscent of typical collagens; however, the protein is not labile to clostridial collagenase, nor does collagen cross-react with antibodies raised against the mussel protein. Unlike typical collagens, Gly probably occurs only at every 4th or 5th residue in this unusual mussel protein. PMID:2481690
Waite, J H; Hansen, D C; Little, K T
Populations undergo physiological adaptations in response to environmental stressors. Our 5-year bio-monitoring study of the Bronx River Estuary demonstrates comparatively low dissolved oxygen concentrations in this urbanized watershed. Additionally, our current results establish altered hormonal levels, resulting from endocrine disruption, in Geukensia demissa (Atlantic ribbed mussel) from the Bronx River Estuary. No studies have yet investigated a correlation between low dissolved oxygen and endocrine disruption in field-collected bivalves. Testosterone, estradiol, and progesterone levels were collected from male and female mussels in the oxygen depleted Bronx River and well-oxygenated Greenwich Cove. Bronx River mussels exhibited higher testosterone levels and lower estradiol levels than Greenwich Cove mussels. The resulting abnormal hormonal ratio seems to indicate that environmental conditions in the Bronx River facilitate an allosteric inhibition of the cytochrome P450 aromatase enzyme, which aids conversion of testosterone to estradiol. Low progesterone levels suggest that Bronx River mussels are experiencing a delay in sexual maturation, and morphometric data show a stalling of shell and tissue growth. To confirm that the mussels collected from both sites are the same species, the universal mitochondrial cytochrome c oxidase subunit I gene was analyzed, through DNA barcoding. Minimal sequential heterogeneity confirmed the mussels are the same species. Such findings suggest intraspecific divergence in various endocrine processes, resulting from environmentally induced stress. PMID:24813673
Halem, Zachery M; Ross, Dustin J; Cox, Rachel L
We studied particle selection in the ribbed mussel Geukensia demissa, an important suspension-feeding inhabitant of estuaries and intertidal zones of salt marshes along the Atlantic coast of North America. Adult mussels were fed on several mixtures of microalgal cultures (1) in exponential or (2) in stationary phase of growth, and the proportional occurrence of algal species in pseudofeces was examined by flow cytometry. The Eastern oyster, Crassostrea virginica, was chosen as a reference. Results showed that both mussels and oysters were able to selectively ingest or reject our experimental microalgae. Moreover, the pre-ingestive particle selection was affected by microalgal growth phase, particularly in mussels. For instance, the sorting efficiency index increased significantly in mussels fed with a blend made of Nitzschia closterium, Isochrysis sp. and Tetraselmis suesica harvested in stationary growth phase, as compared to the same blend made with microalgae in exponential growth phase. Isochrysis sp. and T. suesica were preferentially ingested by both bivalves whereas N. closterium, was preferentially rejected in pseudofeces. These results demonstrate particle selection in ribbed mussel and underline the effect of algae growth phase on the sorting mechanisms.
Pales Espinosa, Emmanuelle; Allam, Bassem; Ford, Susan E.
During aerial exposure (emersion), most sessile intertidal invertebrates experience cellular stress caused by hypoxia, and the amount and types of hypoxia-induced stress will differ as exposure time increases, likely leading to altered metabolic responses. We examined proteomic responses to increasing emersion times and decreasing recovery (immersion) times in the mussel Geukensia demissa, which occurs in salt marshes along the east coast of North America. Individuals are found above mean tide level, and can be emersed for over 18 h during spring tides. We acclimated mussels to full immersion at 15°C for 4 weeks, and compared changes in gill protein expression between groups of mussels that were continually immersed (control), were emersed for 6 h and immersed during recovery for 18 h (6E/18R), were emersed for 12 h and recovered for 12 h (12E/12R), or were emersed for 18 h with a 6 h recovery (18E/6R). We found clear differences in protein expression patterns among the treatments. Proteins associated with anaerobic fermentation increased in abundance in 6E/18R but not in 12E/12R or 18E/6R. Increases in oxidative stress proteins were most apparent in 12E/12R, and in 18E/6R changes in cytoskeletal protein expression predominated. We conclude that G. demissa alters its strategy for coping with emersion stress over time, relying on anaerobic metabolism for short- to medium-duration exposure, but switching to an air-gaping strategy for long-term exposure, which reduces hypoxia stress but may cause structural damage to gill tissue. PMID:24501137
Fields, Peter A; Eurich, Chris; Gao, William L; Cela, Bekim
Two indigenous ribbed mussel (Geukensia demissa) populations were sampled approximately every four weeks during 1997 to investigate the relationships among concentrations of total lipid, lipid classes, and polychlorinated biphenyls (PCBs). One population was located in a highly c...
Abstract:Lecanora demissa (Körb.) Zahlbr. is a crustose, lobate lichen that produces soredia and conidiomata but no apothecia. Its placement in Lecanora has long been questioned but nothing better has been proposed. We have studied the nuclear rDNA of the ITS regions and the SSU of L. demissa. In an alignment of the ITS regions of several representatives of Lecanora s.
Ulf Arup; Martin Grube
This Fisheries and Oceans Canada website provides information regarding shellfish biotoxins and shellfish closures in the Pacific region of Canada. Links are provided to information about amnesic shellfish poisoning biotoxins, paralytic shellfish poisoning biotoxins, other shellfish biotoxins, and shellfish contamination closures.
Fish and shellfish are important in the American diet and economy. Nearly $27 billion are spent each year in the United States\\u000a on seafood products. Fish and shellfish are also important causes of food hypersensitivity. In fact, shellfish constitute\\u000a the number one cause of food allergy in the American adult. During the past decade, much has been learned about allergens
Laurianne G. Wild; Samuel B. Lehrer
This web page discusses Diarrheic Shellfish Poisoning (DSP), a gastrointestinal illness caused by the consumption of contaminated shellfish. The associated toxins are okadaic acid and its derivatives, which are produced by the marine dinoflagellate Dinophysis. This page describes clinical presentation (symptoms) of DSP, diagnosis, management and treatment, chemical structure of okadaic acid, molecular mechanism of action, and references.
This EnviroFacts informational page discusses how red tide develops and how people might be affected by this phenomenon. It covers physical damage, oxygen depletion, direct poisoning, and indirect poisoning including paralytic shellfish poisoning (PSP), diarrhetic shellfish poisoning (DSP), neurotoxic shellfish poisoning (NSP), amnesiac shellfish poisoning (ASP), and aerosol toxins. The page concludes with a discussion of red tide and the role of the sea fisheries research institute.
Maneveldt, Gavin W.; Matthews, Sue; Pitcher, Grant; Van Der Vyver, Irma
Neurotoxic shellfish poisoning (NSP) is caused by consumption of molluscan shellfish contaminated with brevetoxins primarily produced by the dinoflagellate, Karenia brevis. Blooms of K. brevis, called Florida red tide, occur frequently along the Gulf of Mexico. Many shellfish beds in the US (and other nations) are routinely monitored for presence of K. brevis and other brevetoxin-producing organisms. As a result, few NSP cases are reported annually from the US. However, infrequent larger outbreaks do occur. Cases are usually associated with recreationally-harvested shellfish collected during or post red tide blooms. Brevetoxins are neurotoxins which activate voltage-sensitive sodium channels causing sodium influx and nerve membrane depolarization. No fatalities have been reported, but hospitalizations occur. NSP involves a cluster of gastrointestinal and neurological symptoms: nausea and vomiting, paresthesias of the mouth, lips and tongue as well as distal paresthesias, ataxia, slurred speech and dizziness. Neurological symptoms can progress to partial paralysis; respiratory distress has been recorded. Recent research has implicated new species of harmful algal bloom organisms which produce brevetoxins, identified additional marine species which accumulate brevetoxins, and has provided additional information on the toxicity and analysis of brevetoxins. A review of the known epidemiology and recommendations for improved NSP prevention are presented.
Watkins, Sharon M.; Reich, Andrew; Fleming, Lora E.; Hammond, Roberta
This web page discusses Amnesic Shellfish Poisoning, a newly identified marine toxin disease associated with blooms of the diatom Pseudonitschia pungens. The page describes clinical presentation (symptoms), diagnosis, management and treatment, chemical structure of domoic acid, molecular mechanism of action, and references.
The aquaculture industry is growing, and shellfish are gaining importance as a food source. Yet shellfish diseases continue to decimate stocks and hamper production. This booklet highlights some of the recent Sea Grant-supported work in shellfish patholog...
This National SeaGrant (PDF) publication discusses Red Tide, Paralytic Shellfish Poisoning (PSP), Domoic Acid, and Amnesiac Shellfish Poisoning (ASP). It includes a description of symptoms as well as a contact number for the shellfish harvest hotline.
SeaGrant; Oregon State University; NOAA
French coastal water pollution has necessitated construction of shellfish depuration stations. Physical and chemical studies indicated that while ozone and bromates are absent from ozonized seawater, there is an increase in dissolved oxygen and residual oxidants. Water quality, temperature, and variations in shellfish filtration capacities appear to govern oxidant longevity.
Y. Fauvel; G. Pons
This report covers the public health aspects of establishing effective oyster production for export to the United States. It includes a review of oyster culture and production, shellfish administration and control activities, shellfish legislation, incide...
C. B. Kelly R. J. Hammerstrom J. B. Engle S. J. Kim K. M. Bae
This USA Today article provides very general information about the 2005 red tide outbreak off the Gulf of Maine. It offers possible explanations for the outbreak, what is being done to protect people from hazards related to red tide, and how the outbreak has devastated local shellfishers.
Hampson, Rick; Today, Usa
Recognition of the limited biological resources and the increasing environmental pollution has emphasised the need for better utilisation of by-products from the fisheries. Currently, the seafood industry is dependent on the processing of the few selected fish and shellfish species that are highly popular with consumers but, from economic and nutritional points of view, it is essential to utilise the
Fabienne Guérard; Daniel Sellos; Yves Le Gal
Contents: Out of an Inland Sea; physical setting, water quality, biology, conflicting uses, markets, management); Oyster; Clams; Musles and other mollusks; Crabs; Shrimp and other nonmolluscan shellfish; Seaweeds.
D. P. Cheney T. F. Mumford
Diarrhetic shellfish poisoning is a gastrointestinal illness caused by consumption of bivalves contaminated with dinophysistoxins. We report an illness cluster in the United States in which toxins were confirmed in shellfish from a commercial harvest area, leading to product recall. Ongoing surveillance is needed to prevent similar illness outbreaks. PMID:23876232
Lloyd, Jennifer K; Duchin, Jeffrey S; Borchert, Jerry; Quintana, Harold Flores; Robertson, Alison
Presents a literature review of disease outbreaks related to fish and shellfish, covering publications of 1976-77. This review covers the chemical, bacterial, and viral diseases that are transmitted by fish and shellfish. A list of 50 references is also presented. (HM)
Discusses the nature and cause of paralytic shellfish poisoning (PSP). Includes toxic dinoflagellate ecology, taxonomy and life history, and chemistry of the toxins. Recent work with trace metals and directions of future research are also given. (MA)
Dale, Barrie; Yentsch, Clarice M.
The purpose of the present study was to determine the protein quality and digestibility of shellfish commonly consumed in Chile, and to estimate its contribution to the protein needs of the Chilean population. The shellfish studied were chorito (Mytilus edulis chilensis), macha (Mesodesma donacium), loco (Concholepas concholepas), cholga (Aulacomya ater), erizo (Loxechinus albus) and almeja (no specific variety). The NPU method was used to determine protein quality. The percentage of protein adequacy for adult rations was calculated according to FAO/WHO 1973. The contribution of shellfish to the protein availability according to the family income of the Santiago population, was also calculated. Most of the shellfish presented NPU values of about 70; the lowest values were found for loco (54.9) and macha (63.3). The apparent and true digestibility gave an average of 83.6 and 90.4, respectively. The percentage of protein adequacy of habitual rations ranged between 27% (erizo) and 58% (loco). The availability of shellfish protein in relation to total protein increased from 0.4 to 2.5% when income increased. It is concluded therefore, that shellfish protein is, in general, of good quality. Nevertheless, it might be considered of poor influence insofar as fulfilling the protein needs of the population studied, whatever its socioeconomic level. PMID:3834878
Pak, N; Vera, G; Araya, H
The popularity of shellfish has been increasing worldwide, with a consequent increase in adverse reactions that can be allergic or toxic. The approximate prevalence of shellfish allergy is estimated at 0.5-2.5% of the general population, depending on degree of consumption by age and geographic regions. The manifestations of shellfish allergy vary widely, but it tends to be more severe than most other food allergens. Tropomyosin is the major allergen and is responsible for cross-reactivity between members of the shellfish family, particularly among the crustacea. Newly described allergens and subtle differences in the structures of tropomyosin between different species of shellfish could account for the discrepancy between in vitro cross-antigenicity and clinical cross-allergenicity. The diagnosis requires a thorough medical history supported by skin testing or measurement of specific IgE level, and confirmed by appropriate oral challenge testing unless the reaction was life-threatening. Management of shellfish allergy is basically strict elimination, which in highly allergic subjects may include avoidance of touching or smelling and the availability of self-administered epinephrine. Specific immunotherapy is not currently available and requires the development of safe and effective protocols.
...prescribed in 870.111-3, insert the following clause: Purchase of Shellfish (APR 1984) The bidder certifies that oysters, clams, and mussels will be furnished only from plants approved by and operated under the supervision of shellfish...
For feeding purposes shellfish filter large amounts of water but also concentrate infectious agents and toxins that are present in the marine environment either naturally or because of pollution. Thus, the consumption of raw or undercooked shellfish is a substantial source of foodborne poisoning, mostly epidemic and sometimes sporadic. Most of shellfish-borne infectious diseases are linked to fecal contamination of the marine environment; they include: thyphoid fever, salmonellosis, shigellosis, campylobacteriosis, cholera, Norwalk or Norwalk-like gastroenteritis and hepatitis A. In warm climates, shellfish contains naturally occurring halopilic Vibrios and may cause severe sporadic infections (septicemias) among very susceptible consumers (immunocompromised). Shellfish also causes outbreaks of paralytic shellfish poisoning (PSP) and diarrheic shellfish poisoning (DSP) when they are contaminated by toxins produced when Dinophisis, a marine plancton, proliferates. Chemical compounds (heavy metals and organic toxins) that are dumped in the environment (soil, air, and water) also reach shellfish harvesting waters where they are cocentrated. Although acute or chronic effects of the chemical contamination of shellfish have not been clearly documented, the cadmium pollution of some shellfish harvesting waters raises a serious problem. Since it is impossible to prevent completely the contamination of coastal waters by any of the agents cited above, the prevention of shellfish-borne diseases requires monitoring of the marine environment and shellfish flesh (coliform count, Dinophysis toxins, heavy metals...). This surveillance allows the classification of growing areas as suitable or not for harvesting and distribution of shellfish. However, this surveillance is not always sensitive enough. Indicators of fecal pollution are particularly not reliable for shellfish viral contamination. A better knowledge of marine biology, the limitation of coastal waters pollution, improved surveillance, the development of more sensitive indicators, the responsabilisation of the industry and the information of the public on the health hazards associated with shellfish consumption are the key issues for the improvement of shellfish-borne disease prevention. PMID:8966339
Desenclos, J C
During the months of November and December 2005, a harmful algal bloom in the northwestern region of Spain caused the accumulation of different algal toxins in several types of shellfish. Many of the shellfish were contaminated at levels above European regulatory limit, presenting serious risks for human health. The analysis of mussels sampled in the affected areas to search for lipophilic toxins, using liquid chromatography tandem mass spectrometry (LC-MS/MS), showed the presence of free okadaic acid (OA) and dinophysistoxin-2 (DTX-2) as well as esters of these toxins. The results also revealed the presence of minor amounts of 13-desmethyl spirolide C (SPX-1) in the analysed samples, although this toxin has never been reported before in Spain. The combination of different MS modes of operation, just as enhanced MS (EMS) and MS(3) experiments, allowed to confirm the first occurrence of spirolides in Spanish shellfish. PMID:17046040
Villar González, A; Rodríguez-Velasco, M L; Ben-Gigirey, B; Botana, L M
The consumption of shellfish has been associated with viral infections even in cases where shellfish complied with the current regulation, which is based on bacterial analysis. In this study, depuration rates of potential indicators and human viruses have been analysed in order to study the use of complementary parameters for evaluating the microbiological quality of depurated shellfish. Depuration of naturally highly polluted mussels has been evaluated and analyses for Escherichia coli, Clostridium perfringens, somatic coliphages, F-RNA phages and bacteriophages infecting Bacteroides fragilis RYC2056 and HSP40, human adenovirus, enterovirus have been done. Seawater of the depuration tank was disinfected by UV irradiation, ozone and passed through a skimmer and a biological filter. The correct functioning of the depuration tank was monitored by the quantification of total organic carbon (TOC), NH4+ and total aerobic bacteria in the seawater. To study the relation between the bacteriophages and the human viruses analysed, a logistic regression model was applied. F-RNA phages are significantly related to human adenoviruses and enteroviruses. Thus, they can be used as a complementary parameter for evaluating the efficiency of the depuration treatment. Somatic coliphages are also significantly associated with enteroviruses. Bacteriophages infecting B. fragilis HSP40 were analysed by the double-agar-layer (DAL) method, which quantifies infectious viruses, and by nested PCR, which detects the presence of the genome of these phages. The highest sensitivity of the molecular techniques was demonstrated and the results obtained are an indicator of a close relation between positive results by PCR and the presence of infectious viral particles in shellfish. All shellfish samples were negative for human viruses by PCR after 5 days of depuration treatment and the results obtained applying a regression model also showed negative results or nearly for F-RNA phages and bacteriophages infecting B. fragilis RYC2056. Thus, in this specific depuration treatment, 5 days may be necessary to assess the sanitary quality of shellfish. PMID:12076030
Muniain-Mujika, I; Girones, R; Tofiño-Quesada, G; Calvo, M; Lucena, F
We describe a case series of seven patients presenting to an emergency department with symptoms of paralytic shellfish poisoning. They developed varying degrees of nausea, vomiting, diarrhea, weakness, ataxia and paresthesias after eating mussels harvested from a beach near their resort. Four patients were admitted to the hospital, one due to increasing respiratory failure requiring endotracheal intubation and the remainder for respiratory monitoring. All patients made a full recovery, most within 24 hours. The ability to recognize and identify paralytic shellfish poisoning and manage its complications are important to providers of emergency medicine.
Hurley, William; Wolterstorff, Cameron; MacDonald, Ryan; Schultz, Debora
This University of Maryland SeaGrant web page explores outbreaks of paralytic shellfish poisoning (PSP), a problem on both the East and West coasts of the US that is caused by several closely related species in the genus Alexandrium. The page explores the underlying physical mechanisms and localization of PSP outbreaks, as well as economic impact.
Kane, Andrew; Jacobs, Dan; The Aquatic Pathobiology Center, University of Maryland; Maryland SeaGrant
This University of Maryland SeaGrant web page discusses the toxic dinoflagellate Gymnodinium breve and its role in red tide blooms and Neurotoxic Shellfish Poisoning (NSP). The page explores the economic, ecological, and health-related effects of red tide blooms, and the causative accumulation of G. breve into blooms that produce the powerful neurotoxins known as brevetoxins.
Kane, Andrew; Jacobs, Dan; The Aquatic Pathobiology Center, University of Maryland; Maryland SeaGrant
This University of Maryland SeaGrant web page discusses amnesic shellfish poisoning (ASP), also known as domoic acid poisoning (DAP). The page discusses the production of domoic acid by the diatom Pseudo-nitzschia, geographic locations of detection, and economic impacts.
Kane, Andrew; Jacobs, Dan; Seagrant, University O.
The resistance of shellfish poison was evaluated in the presence of 7 chemical disinfectants. Sodium hypochlorite was effective in neutralizing the toxicity of the poison at concentration of 3 parts NaOCl per million (ppm) per microgram of poison at room ...
C. D. Chin
Over the past decade, the shellfish and finfish aquaculture industry has expanded rapidly in coastal British Columbia (BC) Canada. Foreshore and nearshore shellfish and finfish aquaculture leaseholds are sited in close proximity or in direct competition with habitat for wild shellfish. As a result, some wild shellfish harvesters believe shellfish farms are significantly reducing access to beaches and estuarine areas
Alyssa Joyce; Rosaline Canessa
Paralytic shellfish toxins (PSTs) are produced by marine and freshwater microalgae and accumulate in shellfish including mussels, oysters, and scallops, causing possible fatalities when inadvertently consumed. Monitoring of PST content of shellfish is therefore important for food safety, with currently approved methods based on HPLC, using pre- or postcolumn oxidation for fluorescence detection (HPLC-FLD). CE is an attractive alternative for screening and detection of PSTs as it is compatible with miniaturization and could be implemented in portable instrumentation for on-site monitoring. In this study, CE methods were developed for C(4) D, FLD, UV absorption detection, and MS-making this first report of C(4) D and FLD for PSTs detection. Because most oxidized toxins are neutral, MEKC was used in combination with FLD. The developed CZE-UV and CZE-C(4) D methods provide better resolution, selectivity, and separation efficiency compared to CZE-MS and MEKC-FLD. The sensitivity of the CZE-C(4) D and MEKC-FLD methods was superior to UV and MS, with LOD values ranging from 140 to 715 ng/mL for CZE-C(4) D and 60.9 to 104 ng/mL for MEKC-FLD. With the regulatory limit for shellfish samples of 800 ng/mL, the CZE-C(4) D and MEKC-FLD methods were evaluated for the screening and detection of PSTs in shellfish samples. While the CZE-C(4) D method suffered from significant interferences from the shellfish matrix, MEKC-FLD was successfully used for PST screening of a periodate-oxidized mussel sample, with results confirmed by HPLC-FLD. This confirms the potential of MEKC-FLD for screening of PSTs in shellfish samples. PMID:24591173
Keyon, Aemi S Abdul; Guijt, Rosanne M; Gaspar, Andras; Kazarian, Artaches A; Nesterenko, Pavel N; Bolch, Christopher J; Breadmore, Michael C
In order to assess the putative toxigenic risk associated with the presence of fungal strains in shellfish-farming areas, Penicillium strains were isolated from bivalve molluscs and from the surrounding environment, and the influence of the sample origin on the cytotoxicity of the extracts was evaluated. Extracts obtained from shellfish-derived Penicillia exhibited higher cytotoxicity than the others. Ten of these strains were grown on various media including a medium based on mussel extract (Mytilus edulis), mussel flesh-based medium (MES), to study the influence of the mussel flesh on the production of cytotoxic compounds. The MES host-derived medium was created substituting the yeast extract of YES medium by an aqueous extract of mussel tissues, with other constituent identical to YES medium. When shellfish-derived strains of fungi were grown on MES medium, extracts were found to be more cytotoxic than on the YES medium for some of the strains. HPLC-UV/DAD-MS/MS dereplication of extracts from Penicillium marinum and P. restrictum strains grown on MES medium showed the enhancement of the production of some cytotoxic compounds. The mycotoxin patulin was detected in some P. antarcticum extracts, and its presence seemed to be related to their cytotoxicity. Thus, the enhancement of the toxicity of extracts obtained from shellfish-derived Penicillium strains grown on a host-derived medium, and the production of metabolites such as patulin suggests that a survey of mycotoxins in edible shellfish should be considered. PMID:24006923
Geiger, M; Guitton, Y; Vansteelandt, M; Kerzaon, I; Blanchet, E; Robiou du Pont, T; Frisvad, J C; Hess, P; Pouchus, Y F; Grovel, O
... smaller portions. Frequently Asked Questions about Mercury in Fish and Shellfish What is mercury and methylmercury? I' ... of fish. 3. Is there methylmercury in all fish and shellfish? Nearly all fish and shellfish contain ...
Bivalves are ancient animals that feed by filtering large volumes of water. In this way, phytoplankton, bacteria and viruses from the water column are greatly concentrated in the mussels. The hazards associated with the consumption of mussels are thus dependent on the occurrence and composition of toxic algae and human microbial pathogens in the areas where shellfish are grown. Diarrheic shellfish toxins have occurred regularly in Sweden during the past 27 years. Peaks of toxins in mussels are mostly recorded from October to December, but the pattern can differ significantly due to location and year, making it hard to predict toxin levels in mussels. With an expansion of aquaculture and a subsequent increase in seafood consumption, better risk management is needed to minimize the effects on humans of algal toxins and human pathogens. New control strategies that have to be implemented are: i) proper site selection of culture installations; ii) regular and cost-efficient monitoring of algae, bacteria and viruses; iii) new indicators for fecal contamination, suitable for the specific locations where shellfish grow; iv) rapid dissemination of information to the industry and public, including risk assessment and advice on how to cope with the situation. PMID:15865311
Rehnstam-Holm, Ann-Sofi; Hernroth, Bodil
Petroleum hydrocarbon concentrations in seawater, surface sediments and culture shellfish were investigated in shellfish culture area of Sanggou Bay from Jan. to Nov. in 2008. Investigation was conducted on the distribution and variation of petroleum hydrocarbon concentrations in seawater and sediments in the shellfish culture area of Sanggou Bay, as well as on the levels and the differences in petroleum hydrocarbon concentrations among the shellfish species. In addition, the petroleum hydrocarbon pollution status in the three media was evaluated and the effects of accumulated petroleum hydrocarbon in shellfish on the food safety risk were discussed. The results indicated: 1) Petroleum hydrocarbon concentrations in seawater in the shellfish culture area of Sanggou Bay were in the range of 3.61 - 98.21 microg/L; the mean values of petroleum hydrocarbon concentrations in sediments were in the range of 6.75-25.95 mg/kg; petroleum hydrocarbon concentrations in culture shellfish were in the range of 2.14- 42.87 mg/kg; and petroleum hydrocarbon concentrations in shellfish varied largely among different species, with the mean values in the sequence of clam Venerupis variegata > oyster > scallop; 2) Monthly petroleum hydrocarbon concentrations in seawater and surface sediments varied significantly in Sanggou Bay shellfish culture area, with the highest and the lowest values of petroleum hydrocarbon concentrations in seawater that occurred in July and in August, respectively, and with the highest and the lowest values of petroleum hydrocarbon concentrations in surface sediments that occurred in September and in March, respectively; 3) According to the corresponding evaluation criteria, the petroleum hydrocarbon pollution status in surface sediments in Sanggou Bay shellfish culture area was unpolluted but the status in surface seawater was polluted. The culture shellfish was also polluted by petroleum hydrocarbon with different degrees among three species, namely, the pollution degree of clam Venerupis variegata and oyster was more serious than that of scallop Chlamys farreri; 4) The results of linear regression analysis showed that petroleum hydrocarbon concentrations in shellfish had a significantly positive correlation with the concentrations in seawater, while the linear correlation between the concentrations in shellfish and surface sediments was not obvious; and 5) According to the pollution level of petroleum hydrocarbon in shellfish, the food safety risk of three culture shellfishes in Sanggou Bay was relatively low. PMID:22619968
Qiao, Xiang-Ying; Chen, Bi-Juan; Zhou, Ming-Ying; Cui, Zheng-Guo
Voltage-gated sodium channels (VGSCs) play a central role in the generation and propagation of action potentials in excitable neurons and other cells and are targeted by commonly used local anesthetics, antiarrhythmics, and anticonvulsants. They are also common targets of neurotoxins including shellfish toxins. Shellfish toxins are a variety of toxic secondary metabolites produced by prokaryotic cyanobacteria and eukaryotic dinoflagellates in both marine and fresh water systems, which can accumulate in marine animals via the food chain. Consumption of shellfish toxin-contaminated seafood may result in potentially fatal human shellfish poisoning. This article provides an overview of the structure, bioactivity, and pharmacology of shellfish toxins that act on VGSCs, along with a brief discussion on their pharmaceutical potential for pain management.
Zhang, Fan; Xu, Xunxun; Li, Tingting; Liu, Zhonghua
An investigation was conducted to determine the processes involved in the contamination of shellfish by viruses. Results of binding-release studies show that the process involves the attachment of viruses to mucus secreted, and then ingested, by shellfish during feeding. Analysis of the mucus-virus bond involved selective degradation of the mucus and use of chemical agents to block carboxyl and sulfate groups on the mucus. Results obtained indicate that the attachment of virus to mucus is primarily ionic and involves the binding of viral particles to sulfate radicals on the mucopolysaccharide moiety of shellfish mucus.
Di Girolamo, R; Liston, J; Matches, J
Abstract: Narragansett Bay salt marshes of similar geomorphology and hydrology were clustered into low, medium and high human impact based on land use and nutrient inputs. We measured abundance, condition index (CI) and growth of ribbed mussels (Geukensia demissa) from 10 salt ma...
The stable nitrogen isotope ratio in tissue of the ribbed mussel (Geukensia demissa) was investigated as an indicator of the source of nitrogen inputs to coastal salt marshes. Initially, mussels were fed a diet of 15N-enriched algae in the laboratory to determine how the tissue n...
The ribbed mussel, Geukensia demissa, is commonly found associated with the salt marsh cordgrass, Spartina alterniflora. Mussels attach to the basal portion of S. alterniflora stems with strong proteinaceous byssal threads and deposit fecal material on the surrounding sediment as a byproduct of their filter-feeding activity. Here I demonstrate by manipulating mussel densities in the field that the presence of
MARK D. BERTNESS
On June 6, 2011, the Section of Epidemiology (SOE) of the Alaska Division of Public Health was notified of a case of paralytic shellfish poisoning (PSP) in southeast Alaska. In collaboration with local partners, SOE investigated and identified a total of eight confirmed and 13 probable PSP cases that occurred during May--June 2011. Warnings to avoid noncommercially harvested shellfish were broadcast on local radio and television and displayed at beaches and in post offices, government offices, and businesses throughout the region. Commercially harvested shellfish, which are tested for the presence of PSP-causing toxins, were safe. Because the risk for PSP is unpredictable, persons who consume noncommercially harvested Alaskan shellfish should know that they are at risk for PSP, and suspected cases should be reported promptly to SOE to initiate control measures in the affected area. PMID:22089968
When Drakes Estero, which lies within the Point Reyes National Seashore (PRNS) about 25 miles northwest of San Francisco, California, was designated by Congress in 1976 as Potential Wilderness, it contained a commercial shellfish (nonnative oyster and cla...
Excess nutrients in the coastal environment have been linked to a host of environmental problems, and nitrogen reduction efforts have been a top priority of resource managers for decades. The use of shellfish for coastal nitrogen remediation has been proposed, but formal incorporation into nitrogen management programs is lagging. Including shellfish aquaculture in existing nitrogen management programs makes sense from environmental, economic, and social perspectives, but challenges must be overcome for large-scale implementation to be possible. PMID:24506309
Rose, Julie M; Bricker, Suzanne B; Tedesco, Mark A; Wikfors, Gary H
Okadaic acid (OA) and dinophysistoxins-1 and -2 (DTX1, DTX2), the toxins responsible for incidents of diarrhetic shellfish\\u000a poisoning (DSP), can occur as complex mixtures of ester derivatives in both plankton and shellfish. Alkaline hydrolysis is\\u000a usually employed to release parent OA\\/DTX toxins, and analyses are conducted before and after hydrolysis to determine the\\u000a concentrations of nonesterified and esterified toxins. Recent
Erin Doucet; Neil N. Ross; Michael A. Quilliam
Marine organisms such as fish and shellfish are rich sources of structurally diverse bioactive nitrogenous components. Based on emerging evidence of potential health benefits, these components show significant promise as functional food ingredients. Activities including antihypertensive, antioxidant, anti-microbial, anti-coagulant, anti-diabetic, anti-cancer, immunostimulatory, calcium-binding, hypocholesteremic and appetite suppression have been reported. Fish and shellfish waste components contain significant levels of high
Pádraigín A. Harnedy; Richard J. FitzGerald
A plan for developing the shellfish industry of Dukes County should be designed to maximize opportunities for 'off-season'--fall, winter and spring--employment. It should therefore concentrate upon the development of shellfish resources that are accessibl...
G. C. Matthiessen R. C. Toner
...2010-07-01 false Subsistence taking of fish, wildlife, and shellfish: general regulations...PUBLIC LANDS IN ALASKA Subsistence Taking of Fish and Wildlife Â§ 242.25 Subsistence taking of fish, wildlife, and shellfish: general...
The potential for poisoning of humans through their consumption of shellfish which have themselves consumed biotoxin producing marine phytoplankton exists in the UK. Toxins are bio-accumulated within the shellfish flesh allowing them to reach harmful concentrations. This threat is in most part mitigated by monitoring programmes that assess both the presence of potentially harmful phytoplankton and shellfish flesh toxicity. However, the medical profession in the UK remains relatively ignorant of the potential for biotoxin derived shellfish toxicity, preventing quantification of magnitude, frequency, and severity of health effects in the community or the medical significance of more recently discovered toxins. While the current causative species and their toxins are relatively well characterised there remains a lack of understanding of the factors governing the temporal and spatial appearance of harmful phytoplankton. Expansion of shellfish aquaculture is likely both worldwide and in the UK. Better understanding of how harmful phytoplankton interact with their environment to promote the sporadic harmful blooms that we observe is required to underpin risk assessments. PMID:20102579
Davidson, Keith; Bresnan, Eileen
The potential for poisoning of humans through their consumption of shellfish which have themselves consumed biotoxin producing marine phytoplankton exists in the UK. Toxins are bio-accumulated within the shellfish flesh allowing them to reach harmful concentrations. This threat is in most part mitigated by monitoring programmes that assess both the presence of potentially harmful phytoplankton and shellfish flesh toxicity. However, the medical profession in the UK remains relatively ignorant of the potential for biotoxin derived shellfish toxicity, preventing quantification of magnitude, frequency, and severity of health effects in the community or the medical significance of more recently discovered toxins. While the current causative species and their toxins are relatively well characterised there remains a lack of understanding of the factors governing the temporal and spatial appearance of harmful phytoplankton. Expansion of shellfish aquaculture is likely both worldwide and in the UK. Better understanding of how harmful phytoplankton interact with their environment to promote the sporadic harmful blooms that we observe is required to underpin risk assessments.
Summary form only given. Coastal shellfish aquaculture sites are subject to seasonal blooms of toxic microalgae, which may result in restrictions on shellfish harvest due to human health risk. In southeastern Nova Scotia, such toxic episodes are uncommon, although significant numbers of putatively toxigenic phytoplankton are often found in the water column. In most coastal regions where diarrhetic shellfish poisoning
A. Cembella; A. Bauder; N. Lewis; M. Quilliam; J. Lawrence; J. Manuel; U. Lobsiger
The distribution of the concentration of potential indicators of fecal viral pollution in shellfish was analyzed under diverse conditions over 18 months in diverse geographical areas. These microorganisms have been evaluated in relation to contamination by human viral pathogens detected in parallel in the analyzed shellfish samples. Thus, significant shellfish-growing areas from diverse countries in the north and south of
M. Formiga-Cruz; A. K. Allard; A.-C. Conden-Hansson; K. Henshilwood; B. E. Hernroth; J. Jofre; D. N. Lees; F. Lucena; M. Papapetropoulou; R. E. Rangdale; A. Tsibouxi; A. Vantarakis; R. Girones
Currently Angola does not possess a monitoring programme for shellfish contamination with marine biotoxins. Among other potentially toxic microalgae, the presence of Gymnodinium catenatum and Pyrodinium bahamense has been reported at the Angola coast, two species associated worldwide with production of paralytic shellfish poisoning toxins (PSTs). A preliminary assessment of contamination with PSTs was carried out by HPLC with pre-column derivatization in samples of Semele proficua from Luanda Bay and samples of Senilia senilis from Mussulo Bay, collected between June 2007 and February 2008. An unusual profile was found, not matching any of the 10 oxidation products expected from the known hydrophilic PSTs normally reported in marine dinoflagellates: the N-sulfocarbamoyl, the decarbamoyl or the carbamoyl analogues of saxitoxin (STX). Four major compounds were noted, and designated A through D. These were not autofluorescent, additionally A and D presented much stronger response after peroxide oxidation than after periodate oxidation. Fluorescence emission and ultraviolet absorption maxima were similar to oxidation products of STX analogues. Separation carried out in two different C18 columns clearly showed the retention times did not match the oxidation products of standards. In the period of August through October 2007, samples of S. proficua from Luanda Bay presented a strong cross-reaction with a commercial antibody towards STX, in the range of 2600-5800STXequiv./kg, while for the same period levels in the samples of S. senilis from Mussulo Bay ranged between <30 and 340STXequiv./kg. This reaction was directly proportional to the presence of the four unknown fluorescent peaks. The presence of these compounds was very persistent throughout the studied period, in particular in Luanda Bay. This contamination might not be characteristic of a dinoflagellate blooming, but could be related to a persistent cyanobacterial contamination, due to the strong sewage pollution of the area. PMID:19041661
Vale, Paulo; Rangel, Isabel; Silva, Bárbara; Coelho, Paulo; Vilar, Andrea
Several Dinophysis species produce diarrhoetic toxins (okadaic acid and dinophysistoxins) and pectenotoxins, and cause gastointestinal illness, Diarrhetic Shellfish Poisoning (DSP), even at low cell densities (<103 cells·L?1). They are the main threat, in terms of days of harvesting bans, to aquaculture in Northern Japan, Chile, and Europe. Toxicity and toxin profiles are very variable, more between strains than species. The distribution of DSP events mirrors that of shellfish production areas that have implemented toxin regulations, otherwise misinterpreted as bacterial or viral contamination. Field observations and laboratory experiments have shown that most of the toxins produced by Dinophysis are released into the medium, raising questions about the ecological role of extracelular toxins and their potential uptake by shellfish. Shellfish contamination results from a complex balance between food selection, adsorption, species-specific enzymatic transformations, and allometric processes. Highest risk areas are those combining Dinophysis strains with high cell content of okadaates, aquaculture with predominance of mytilids (good accumulators of toxins), and consumers who frequently include mussels in their diet. Regions including pectenotoxins in their regulated phycotoxins will suffer from much longer harvesting bans and from disloyal competition with production areas where these toxins have been deregulated.
Reguera, Beatriz; Riobo, Pilar; Rodriguez, Francisco; Diaz, Patricio A.; Pizarro, Gemita; Paz, Beatriz; Franco, Jose M.; Blanco, Juan
A chemical assay for saxitoxin, the paralytic shellfish poison, has been developed. The technique involves alkaline hydrogen peroxide oxidation of saxitoxin to 8-amino-6-hydroxymethyl-2-iminopurine-3(2H)-propionic acid, the fluorescence of which is measur...
H. A. Bates H. Rapoport
This map represents the Health Status of beaches in the state of Washington. The interactive map allows users to click on counties, water bodies, and beaches to view seasons and limits. The page also includes links to text bulletins regarding beach closures, descriptions of marine biotoxins and associated health effects, and a factsheet of shellfish program publications.
Health, Washington S.
Bivalve shellfish are at constant risk of being exposed to pathogens as a consequence of contamination of the shellfish beds with human or animal waste originating from sewage treatment plants or slurry fertilized fields. Consumption of contaminated oysters and mussels are frequently reported as causes of disease outbreaks caused by norovirus or hepatitis A virus. Other zoonotic pathogens such as hepatitis E virus (HEV), rotavirus (RV) and Salmonella from livestock may also be transmitted to shellfish via this route. In this study, 29 pooled samples from commercial Danish blue mussels were tested for porcine pathogens and indicator bacteria Escherichia coli (E. coli). All samples tested negative for HEV, RV and Salmonella, whereas E. coli and the highly stable porcine circovirus type 2 (PCV2) were detected in eight and 12 samples, respectively. This is the first study to report the detection of PCV2 in commercial mussels. Based on the detection of PCV2 in clean areas with low prevalence of the normally applied fecal indicator E. coli, testing for PCV2 may be a more sensitive and robust specific porcine waste indicator in shellfish harvesting areas. PMID:25016209
Krog, J S; Larsen, L E; Schultz, A C
Background and Objectives In the present study, isolation of Streptomyces associated with marine sponges and its bioactive potential against fish and shellfish pathogens were assessed. The Streptomyces sp. were isolated from the marine sponges namely Callyspongia diffusa, Mycale mytilorum, Tedania anhelans and Dysidea fragilis collected from Vizhinjam port, situated in the South-West coast of India. Materials and Methods The Streptomyces associated with marine sponges were isolated using specific ISP media. The isolates of Streptomyces were characterized for their colony characteristics, morphological properties, physiological and biochemical properties and were tentatively identified. The strains were cultivated on a lab scale level as shake-flask cultures and the crude extracts of the bioactive compounds obtained with ethyl acetate were screened biologically and chemically. By biological screening, the extracts were analyzed for their activity against fish and shellfish pathogens namely Aeromonas hydrophila, Serratia sp. and Vibrio spp, using the disk and agar-well diffusion bioassay method, while by chemical screening the crude culture extracts were analyzed by TLC and UV–Vis spectrophotometer. Results Ninety-four isolates were found to be associated with marine sponges, among them only seven strains showed antagonism against fish and shellfish pathogens. Analysis of morphological, physiological and biochemical characteristics suggested that these strains belonged to the genus Streptomyces. The initial screening of the isolates by spot inoculation method exhibited antibacterial activity against Aeromonas hydrophila. In-vitro screening of the submerge culture extracts showed positive inhibition against the fish and shellfish pathogens namely Aeromonas hydrophila, Serratia sp. and Vibrio spp. The screening of bioactive compounds confirmed the production of polyene substances by UV spectrum, which resulted in absorbance peaks ranging from 225 to 245 nm and TLC analysis yielded Rf values ranging from 0.40 to 0.78. Conclusion The results suggest that the seven Streptomyces strains isolated from marine sponges produce potential antibacterial compounds against fish and shellfish pathogens.
Selvakumar, D; Arun, K; Suguna, S; Kumar, D; Dhevendaran, K
Using a streptavidin-coated well plate, a biotin-labelled anti-gonyautoxin 2/3 monoclonal antibody GT-13A, and a decarbamoyl saxitoxin-peroxidase conjugate, a direct competitive enzyme-linked immunosorbent assay (PSP-ELISA) was developed for monitoring paralytic shellfish poisoning (PSP) toxins in shellfish. This assay is simple to perform and can be completed in approximately 20 min. The PSP-ELISA was compared to the mouse bioassay (MBA) for the detection of PSP toxins in shellfish samples (n=83) collected from the coast of Osaka Prefecture, Japan. When positive and negative results were indicated based on the regulatory limit for PSP toxins (4 mouse unit(MU)/g of shellfish meat), the PSP-ELISA results showed a sensitivity of 100% (25 of 25) and a specificity of 89.7% (52 of 58 samples) compared to the MBA results. These results suggest that the PSP-ELISA could be used as a rapid and simple screening method prior to the MBA. PMID:24874362
Kawatsu, Kentaro; Kanki, Masashi; Harada, Tetsuya; Kumeda, Yuko
A novel extraction and clean-up method has been developed for the determination of paralytic shellfish poisoning (PSP) toxins in shellfish samples. Raw shellfish material was extracted with an acidic acetonitrile/water (80:20, v/v) solution, whilst being homogenised. During the homogenisation the sample extraction solution was cooled with ice water. Subsequently, the extract was frozen at -20 degrees C for at least 4h. During freezing, two layers were formed, only the lower predominantly aqueous layer was used for the determination. The final extract solution was cleaned-up using a combination of Oasis HLB and Carbograph activated carbon SPE columns. The developed extraction and clean-up methods combined with gradient elution liquid chromatography (LC)-mass spectrometry/mass spectrometry (MS/MS) has resulted in a method which can determine the analogs GTX 1-5, C1-2, DcGTX 2-3, DcSTX, Neo, STX in a single analysis with an overall detection limit of 313 microg STXdiHCL-eq./kg shellfish meat. The use of the developed extraction method with post-column high performance liquid chromatography (HPLC) with fluorescence detection (FLD) method provided an overall limit of detection of 89 microg STXdiHCL-eq./kg shellfish meat for the same toxins. Both post-column HPLC-FLD and LC-MS/MS was used to investigate the Norwegian PSP toxin profile. It was found that the PSP toxins could be detected in shellfish samples from the Norwegian coastline for 10 months of the year, from March till December. The toxin profile consisted mainly of the carbamate toxins, GTX 1-4, Neo and STX, in terms of both concentrations and contribution to the overall toxicity. In addition, several of the n-sulfo-carbamoyl toxins were either detected in the samples at relatively low concentrations or their presence in the samples were indicated but could not be confirmed by the post-column HPLC-FLD and LC-MS/MS analyses. PMID:18619989
Sayfritz, S J; Aasen, J A B; Aune, T
This photograph shows Texas hornshell mussels (Popenaias popeii) marked with shellfish tags, an example of marks used on organisms for long-term studies of survival and movement. In addition, growth rings, thought to be produced annually, are clearly visible on these mussels. Flat shellfish tags (white with numbers) have been glued near the umbo on the hinge line of the mussel. The umbo is a light-colored raised area at the top of mussels in this photo. This mussel species is a priority 2 candidate for federal endangered species listing, listed as endangered by the State of New Mexico, and is listed as critically endangered on IUCNÃÂs red list. Mussels are amongst North AmericaÃÂs most endangered freshwater taxa.
A simple, rapid, high-performance liquid chromatographic experiment suitable for undergraduate students is described for determining amnesic shellfish poison in mussels. The poison itself is an unusual naturally occurring amino acid, domoic acid, that has been found in seafood, particularly shellfish, worldwide. The symptoms of poisoning include amnesia (memory loss), loss of balance, mental confusion, nausea, vomiting, diarrhea, coma, and in extreme cases death. The domoic acid is extracted from homogenized mussel tissue by boiling in water for 5 minutes. The homogenate is cooled and centrifuged, and an aliquot of the supernatant is diluted and analyzed by isocratic HPLC using a C18 column and an acetonitrile-water mobile phase at pH 2.5 with UV detection at 242 nm.
Among the thousands of unicellular phytoplankton species described in the sea, some frequently occurring and bloom-forming marine dinoflagellates are known to produce the potent neurotoxins causing paralytic shellfish poisoning. The natural function of these toxins is not clear, although they have been hypothesized to act as a chemical defence towards grazers. Here, we show that waterborne cues from the copepod Acartia tonsa induce paralytic shellfish toxin (PST) production in the harmful algal bloom-forming dinoflagellate Alexandrium minutum. Induced A. minutum contained up to 2.5 times more toxins than controls and was more resistant to further copepod grazing. Ingestion of non-toxic alternative prey was not affected by the presence of induced A. minutum. The ability of A. minutum to sense and respond to the presence of grazers by increased PST production and increased resistance to grazing may facilitate the formation of harmful algal blooms in the sea.
Selander, Erik; Thor, Peter; Toth, Gunilla; Pavia, Henrik
Among the thousands of unicellular phytoplankton species described in the sea, some frequently occurring and bloom-forming marine dinoflagellates are known to produce the potent neurotoxins causing paralytic shellfish poisoning. The natural function of these toxins is not clear, although they have been hypothesized to act as a chemical defence towards grazers. Here, we show that waterborne cues from the copepod Acartia tonsa induce paralytic shellfish toxin (PST) production in the harmful algal bloom-forming dinoflagellate Alexandrium minutum. Induced A. minutum contained up to 2.5 times more toxins than controls and was more resistant to further copepod grazing. Ingestion of non-toxic alternative prey was not affected by the presence of induced A. minutum. The ability of A. minutum to sense and respond to the presence of grazers by increased PST production and increased resistance to grazing may facilitate the formation of harmful algal blooms in the sea. PMID:16769640
Selander, Erik; Thor, Peter; Toth, Gunilla; Pavia, Henrik
For the past 11 years the annual Shellfish Biology Seminar at Milford CT has provided a unique forum for aquaculture scientists and industry officials to exchange information about estuaries facing increased pollution problems, especially Long Island Sound and the Great South Bay. Because these two areas are so rich in productivity and diversity, fish and shellfish farmers utilize their waters, shellfish beds, and shore land for hatcheries and grow-out facilities. These individuals seek better management of the coastal estuarine environment and its resources, providing a working example of environmental stewardship. In aquaculture, good science is required to understand the complex variables and interaction of estuarine currents, tides, temperature, and cycles of reproduction. Aquaculturists are beginning to understand the need for specific nutrients and how the wastes of one species can be utilized for enhanced production of another species. Over the years, this meeting has formed an amalgam of both the aquaculture industry and research scientists where both groups foster mutual environmental concern. Science is able to focus on the theoretical aspects of pollutant damage. while the aquaculture industry is able to define the problem and need for assistance to eliminate pollutants from their crops—shellfish and finfish. Overfishing is not an issue at these meetings, as the group accepts the damage already done to wild resources and seeks new technologies to grow food sources under controlled and stable market conditions. Therefore, it could be said that the seminar serves as a meeting ground where the theoretical knowledge of scientific study finds practical application in the industry and is fueled by the needs of that industry. This ideal blend of the two groups produces better management of the resource and a safer environment—the goal of stewardship.
Blogoslawski, Walter J.
Viral contamination in oyster and mussel samples was evaluated after a massive storm with hurricane wind named "Xynthia tempest" destroyed a number of sewage treatment plants in an area harboring many shellfish farms. Although up to 90% of samples were found to be contaminated 2 days after the disaster, detected viral concentrations were low. A 1-month follow-up showed a rapid decrease in the number of positive samples, even for norovirus. PMID:22344664
Grodzki, Marco; Ollivier, Joanna; Le Saux, Jean-Claude; Piquet, Jean-Côme; Noyer, Mathilde; Le Guyader, Françoise S
An interlaboratory trial was conducted to validate photostimulated luminescence (PSL) detection of irradiated shellfish. Five species of shellfish (Nephrops norvegicus, mussels, black tiger prawns, brown shrimps, and king scallops) were presented blind as nonirradiated and irradiated to 0.5 and 2.5 kGy. Precharacterization analysis of each product and treatment was performed on both whole (including shell) and intestinal samples. The results for whole samples (including shell) confirmed that the method was able to distinguish between nonirradiated and irradiated samples, regardless of dose. Intestinal data have identified that the method is dependent on the quantity and sensitivity of grits present within the intestinal tract, which can be assessed using calibration by normalization to 1 kGy. Five laboratories returned both initial screening and calibrated data and sample classification. All laboratories correctly identified all irradiated products using the screening criteria. There were no false positives. The results confirm the validity of the PSL method for shellfish, which has been adopted as a European standard method and by the Codex Alimentarius Commission. Calibration is required where only intestinal material is available. For whole samples with shell, screening alone is sufficient. PMID:14632401
Sanderson, David C W; Carmichael, Lorna A; Fisk, Saffron
Water pollution represents a threat of increasing importance to human health. Bivalve mollusks are filter-feeding organisms that can accumulate chemical and microbiological contaminants in their tissues from very low concentrations in the water or sediments. Consumption of contaminated shellfish is one of the main causes of seafood poisoning. Thus, marine bivalves are normally depurated in sterilized seawater for 48h to allow the removal of bacteria. However, this depuration time might be insufficient to eliminate chemical contaminants from their tissues. We have developed a novel technology that accelerates up to fourfold the excretion rate of xenobiotics in bivalves by treatment with the antioxidant and glutathione (GSH) pro-drug N-acetylcysteine (NAC) during the depuration period. NAC improved dose-dependently the detoxification of the organophosphate (OP) pesticide fenitrothion in the mussel Mytilus galloprovincialis, diminishing its levels up to nearly a hundred fold compared to conventional depuration, by enhancing the glutathione S-transferase (GST) activity and inducing the GSH anabolism (GSH synthesis and reduction by glutathione reductase). Notably, this induction in GSH anabolism and GST activity was also observed in uncontaminated bivalves treated with NAC. As the GSH pathway is involved in the detoxification of many pollutants and biotoxins from harmful algal blooms, we validated this proof of principle in king scallops (Pecten maximus) that naturally accumulated the amnesic shellfish poisoning (ASP) toxin domoic acid. We illustrate here a method that enhances the elimination of organic contaminants in shellfish, opening new avenues of depuration of marine organisms. PMID:24880785
Peña-Llopis, Samuel; Serrano, Roque; Pitarch, Elena; Beltrán, Eduardo; Ibáñez, María; Hernández, Félix; Peña, Juan B
Paralytic shellfish poisoning (PSP) is a potentially fatal human health condition caused by the consumption of shellfish containing high levels of PSP toxins. Toxin extraction from shellfish and from algal cultures for use as standards and analysis by alternative analytical monitoring methods to the mouse bioassay is extensive and laborious. This study investigated whether a selected MAb antibody could be coupled to a novel form of magnetic microsphere (hollow glass magnetic microspheres, brand name Ferrospheres-N) and whether these coated microspheres could be utilized in the extraction of low concentrations of the PSP toxin, STX, from potential extraction buffers and spiked mussel extracts. The feasibility of utilizing a mass of 25 mg of Ferrospheres-N, as a simple extraction procedure for STX from spiked sodium acetate buffer, spiked PBS buffer and spiked mussel extracts was determined. The effects of a range of toxin concentrations (20-300 ng/mL), incubation times and temperature on the capability of the immuno-capture of the STX from the spiked mussel extracts were investigated. Finally, the coated microspheres were tested to determine their efficiency at extracting PSP toxins from naturally contaminated mussel samples. Toxin recovery after each experiment was determined by HPLC analysis. This study on using a highly novel immunoaffinity based extraction procedure, using STX as a model, has indicated that it could be a convenient alternative to conventional extraction procedures used in toxin purification prior to sample analysis.
Devlin, Raymond; Campbell, Katrina; Kawatsu, Kentaro; Elliott, Christopher
Paralytic shellfish poisoning (PSP) is a potentially fatal human health condition caused by the consumption of shellfish containing high levels of PSP toxins. Toxin extraction from shellfish and from algal cultures for use as standards and analysis by alternative analytical monitoring methods to the mouse bioassay is extensive and laborious. This study investigated whether a selected MAb antibody could be coupled to a novel form of magnetic microsphere (hollow glass magnetic microspheres, brand name Ferrospheres-N) and whether these coated microspheres could be utilized in the extraction of low concentrations of the PSP toxin, STX, from potential extraction buffers and spiked mussel extracts. The feasibility of utilizing a mass of 25 mg of Ferrospheres-N, as a simple extraction procedure for STX from spiked sodium acetate buffer, spiked PBS buffer and spiked mussel extracts was determined. The effects of a range of toxin concentrations (20-300 ng/mL), incubation times and temperature on the capability of the immuno-capture of the STX from the spiked mussel extracts were investigated. Finally, the coated microspheres were tested to determine their efficiency at extracting PSP toxins from naturally contaminated mussel samples. Toxin recovery after each experiment was determined by HPLC analysis. This study on using a highly novel immunoaffinity based extraction procedure, using STX as a model, has indicated that it could be a convenient alternative to conventional extraction procedures used in toxin purification prior to sample analysis. PMID:22069687
Devlin, Raymond; Campbell, Katrina; Kawatsu, Kentaro; Elliott, Christopher
Estuarine intertidal flats are often exploited by humans and waders since they provide food, particularly shellfish. This raises important conservation issues. Waders can be affected by shellfishing activities in multiple ways, such as a reduction of the available shellfish, disturbance by the presence of shellfishers on their feeding areas, and changes in micro-habitat, due to sediment reworking. In this study we quantified the impact of traditional shellfishing on waders in the Tagus estuary. Particular attention was given to hand-raking of clams Scrobicularia plana, which constitutes most of the consumed food by waders. Shellfishers did not cause a relevant depletion of clams for waders; they removed less than 0.3% of its total production and focused on size classes that were usually not taken by birds. Hand-raking caused temporary changes in the vertical distribution and availability of invertebrate prey in the sediment. However, this did not affect the bird's feeding rates, presumably because prey availability remained above the threshold at which intake rates are expected to decline. The presence of shellfishers in the birds foraging areas potentially affects waders by keeping them away from foraging areas, but even the most affected species lost less than 10% of their foraging grounds due to this factor. Overall, we conclude that the current low harvesting levels of shellfishing are compatible with the preservation of the estuary as a key site for waders. Nevertheless, simulations showed that traditional shellfishing could have much greater potential to affect waders through disturbance than through prey removal. The results for the Tagus show that even small harvest rates, representing a negligible loss of food for waders and potentially considered sustainable by shellfish managers, could have a great impact on waders due to increased disturbance. This effect of disturbance likely occurs in most estuaries and should be taken into consideration when planning shellfish harvesting in important wader feeding areas.
Dias, Maria P.; Peste, Filipa; Granadeiro, José P.; Palmeirim, Jorge M.
One hundred eighty-four serologically confirmed cases of hepatitis A were reported in eastern Spain in 1999. A matched case-control study implicated imported coquina clams complying with European Union shellfish standards as the source of infection; this implication was confirmed by the detection by reverse transcription- PCR of hepatitis A virus (HAV) RNA in shellfish samples. In spite of the recognized
Gloria Sanchez; Rosa M. Pinto; Hermelinda Vanaclocha; Albert Bosch
We describe the evaluation of a nested reverse transcriptase PCR (RT-PCR) procedure for the detection of small round-structured viruses (SRSVs) in molluscan shellfish and the application of this assay for the detection of SRSVs in commercially produced shellfish and in shellfish implicated in outbreaks of gastroen- teritis. The range of virus strains detected and the sensitivity of detection were evaluated
J. GREEN; K. HENSHILWOOD; C. I. GALLIMORE; D. W. G. BROWN; D. N. LEES
Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator, male-specific B+ coliphages, have been investigated as viral indicators of faecal contamination that may provide source-specific information for impacted environmental waters. This study compared the distribution of E. coli and F+ RNA bacteriophages in shellfish grown in harvesting areas of Greece and also examined the presence and proportions of the different subgroups of F+ RNA coliphages in shellfish. F+ RNA bacteriophages were present in shellfish at higher concentrations than E. coli. Elevated numbers of F+ RNA bacteriophages observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in Greece. The majority of F+ RNA coliphages detected in shellfish samples belonged to group IV which indicated the possible presence of animal faecal material in sample harvesting areas. Phages of groups II and III (human waste and human faecal material, respectively) were present at low levels. Finally, 8% of the phages hybridised were found to belong to group I. The presence of group IV showed seasonal distribution (more in winter, less in summer) whereas the other groups did not show any difference. Monitoring of F+ coliphage subgroups may indicate the presence and major sources of microbial inputs to surface waters; however, environmental effects on the relative occurrence of different groups need to be considered. PMID:17037156
Vantarakis, A; Venieri, D; Komninou, G; Papapetropoulou, M
Speciation of arsenic has received significant attention over the past 20 years in both mechanistic and exposure assessment research. Because the toxicity of arsenic is related to its oxidation state and its chemical forms, the determination of the total arsenic contents in a sample is not adequate to allow its impact on living organisms to be estimated. The inorganic arsenic species, arsenite (As3+) and arsenate (As5+), have been classified as carcinogenic and the methylated forms, monomethyl arsonic acid (MMA) and dimethyl arsinic acid (DMA) have recently been identified as cancer promoters. The highly methylated compounds like as arsenobetaine (AsB) and arsenocholine (AsC) are considered to be nontoxic. Although organisms in marine environment contain high amounts of total arsenic (ppm level), it is not usually present as inorganic arsenic or simple methylated forms well known as one of the toxic species. Arsenobetaine is the dominant species in marine animals and arsenosugars are most abundant in marine algae. This study aims to clarify those arsenic species present in the whole body of eleven different shellfishes from Korea. And those arsenic species were separated and measured by characterization using high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) coupled system. The separation of arsenic species was achieved on anion exchange column and cation exchange column using phosphate and pyridine eluent, respectively. The ultrasonic extraction was employed for extraction of arsenic from whole body of shellfishes. The method was validated by analyzing three certified reference materials (DORM-2, TORT-2, 1566b). Total arsenic concentrations ranged from 0.1 mg/kg dry mass to 21.7 mg/kg dry mass. Most marine shellfishes contained higher arsenobetaine and arsenocholine with the exception of two shellfishes living in river. The lower amounts of inorganic arsenic species were also found in the some sample extracts. Detection of inorganic arsenic can be explained by the conversion of inorganic arsenic to organic arsenic compounds in digestion system in the body may be occurring.
Yoon, C.; Yoon, H.
Paralytic shellfish poisoning (PSP) toxins are highly toxic natural compounds produced by dinoflagellates commonly present in marine phytoplankton. Shellfish accumulate PSP toxins during toxic algal blooms. PSP poses a significant public health threat and economic loss to the shellfish industry. For this reason, many methods have been developed to analyse PSP toxins in suspected toxic shellfish samples. In our study,
Chih Yu Chen; Hong Nong Chou
An international interlaboratory trial was conducted using thermoluminescence for the detection of irradiated shellfish, aimed at validating the method for routine use. Nephrops norvegicus, mussels, brown shrimps, black tiger prawns, and king scallops were presented as nonirradiated and irradiated to 0.5 and 2.5 kGy. The protocol called for the use of 3 preparation methods: extraction of silicates from whole shellfish by acid hydrolysis and physical separation, and of carbonates from powdered shells. Homogeneity was tested on each product and each treatment. Results verified that all methods were able to distinguish between nonirradiated and irradiated samples regardless of dose. Silicate methods produced better discrimination than powdered shell, and acid hydrolysis showed some evidence of better separation between the 2 doses than the physical method. Participants received each product in each treatment category for blind analysis. Six participants returned results for acid hydrolysis, 7 for physical separation, and 5 for the powdered shell method. Their results confirmed the homogeneity testing. Qualitative results gave 100% correct classification for both silicate methods and 85.3% for powdered shell. Silicate methods are therefore preferable unless only shell is available. Overall, the results confirmed the case for validation. PMID:14632400
Sanderson, David C W; Carmichael, Lorna A; Fisk, Saffron
Mangrove oysters (Crassostrea rhizophorae) were screened for enteric viruses. For 18 months oysters were collected from Cano Boqueron, a tropical mangrove lagoon on the southwest coast of Puerto Rico. This popular tourist resort has two primary sewage treatment plants which service 158 single family cabanas. In spite of the heavy seasonal input of sewage to Cano Boqueron and high densities of fecal coliform bacteria, enteric viruses were not detected in shellfish meat. Because no viruses were detected in the oysters, a virus survival study was performed. Poliovirus type 1 was placed in diffusion chambers in situ at two sites in Cano Boqueron. More than 95% of the poliovirus inactivation occurred within 24 h. Virus inactivation was significantly different by site, indicating different inactivation rates within the lagoon. Chamber studies done simultaneously with Escherichia coli did not reveal differences between sites. It is suggested that the sewage effluent had an antiviral effect in the absence of an antibacterial effect. This study demonstrates the importance for establishing microbial contamination standards for shellfish growing waters in the tropics based upon in situ studies with tropical species, e.g. mangrove oyster.
Lopez de Cardona, I.; Bermudez, M.; Billmire, E.; Hazen, T.C. [Univ. of Puerto Rico, Rio Piedras (Puerto Rico)
It has been almost a decade since a previously unknown human toxic syndrome, azaspiracid poisoning (AZP), emerged as the cause of severe gastrointestinal illness in humans after the consumption of mussels (Mytilus edulis). Structural studies indicated that these toxins, azaspiracids, were of a new unprecedented class containing novel structural features. It is now known that the prevalent azaspiracids in mussels are AZA1, AZA2 and AZA3, which differ from each other in their degree of methylation. Several hydroxylated and carboxylated analogues of the main azaspiracids have also been identified, presumed to be metabolites of the main toxins. Since its first discovery in Irish mussels, the development of facile sensitive and selective LC-MS/MS methods has resulted in the discovery of AZA in other countries and in other species. Mice studies indicate that this toxin class can cause serious tissue injury, especially to the small intestine, and chronic exposure may increase the likelihood of the development of lung tumours. Studies also show that tissue recovery is very slow following exposure. These observations suggest that AZA is more dangerous than the other known classes of shellfish toxins. Consequently, in order to protect human consumers, proper risk assessment and regulatory control of shellfish and other affected species is of the utmost importance. PMID:20026101
Furey, Ambrose; O'Doherty, Sinead; O'Callaghan, Keith; Lehane, Mary; James, Kevin J
The GPTT virus RNA extraction method, originally developed for extraction of human norovirus and hepatitis A virus RNAs from contaminated shellfish, was evaluated for extraction of RNA from Aichi virus strain A846/88 (AiV), coxsackievirus strains A9 (CAV9) and B5 (CBV5), murine norovirus (strain MNV-1), and the norovirus surrogate, feline calicivirus (FCV) strain KCD, for the purpose of RT-PCR detection within seeded oyster (Crassostrea virginica) extracts. The RT-PCR equivalent sensitivities observed within seeded oysters as compared to virus stocks were 0.68, 6.8, 26, 5.6, and 14.5 RT-PCR(50) units when assaying 10% of total RNA extracted from seeded oyster extracts for CAV9, CBV5, AiV, FCV, and MNV-1, respectively. For oysters exposed to virus-contaminated seawater, the detection equivalent sensitivities observed were 680, 68, 2600, 560, and 14.5 RT-PCR(50) for CAV9, CBV5, AiV and FCV, and MNV-1, respectively. These results indicate that the GPTT method can be used as a general viral RNA extraction method for multiple picornaviruses and caliciviruses that could potentially contaminate shellfish. PMID:17184849
Kingsley, David H
Paralytic shellfish poisoning (PSP), due to saxitoxin and related compounds, typically results from the consumption of filter-feeding molluscan shellfish that concentrate toxins from marine dinoflagellates. In addition to these microalgal sources, saxitoxin and related compounds, referred to in this review as STXs, are also produced in freshwater cyanobacteria and have been associated with calcareous red macroalgae. STXs are transferred and bioaccumulate throughout aquatic food webs, and can be vectored to terrestrial biota, including humans. Fisheries closures and human intoxications due to STXs have been documented in several non-traditional (i.e. non-filter-feeding) vectors. These include, but are not limited to, marine gastropods, both carnivorous and grazing, crustacea, and fish that acquire STXs through toxin transfer. Often due to spatial, temporal, or a species disconnection from the primary source of STXs (bloom forming dinoflagellates), monitoring and management of such non-traditional PSP vectors has been challenging. A brief literature review is provided for filter feeding (traditional) and non-filter feeding (non-traditional) vectors of STXs with specific reference to human effects. We include several case studies pertaining to management actions to prevent PSP, as well as food poisoning incidents from STX(s) accumulation in non-traditional PSP vectors.
Deeds, Jonathan R.; Landsberg, Jan H.; Etheridge, Stacey M.; Pitcher, Grant C.; Longan, Sara Watt
Two marine mussels, Geukensia demissa (Dillwyn) and Mytilus edulis (L.) collected in 1990 in Old Silver Beach, Falmouth, Massachusetts, incorporated nitrogen when fed 15N-labelled organic aggregates produced from dissolved organic nitrogen released by the brown sea-weed Fucus vesiculosis. Uptake of 15N on the aggregate diet was linear over the course of 24 h, and unincorporated 15N was eliminated from the
M. Alber; I. Valiela
Noroviruses (NoVs) are commonly occurring pathogens that cause gastroenteritis. Outbreaks of viral diseases have often been ascribed to the consumption of contaminated shellfish. Our objective was to evaluate the presence and contamination levels of NoV in shellfish sold at seafood markets in China. We tested 840 shellfish samples (Crassostrea gigas, Mytilus edulis, Azumapecten farreri, SinoNoVacula constricta, Scapharca subcrenata, Ruditapes philippinarum) that were collected from seven cities around the Yellow and Bohai Seas in China between December 2009 and November 2011. We used real-time RT-PCR to detect NoV in purified concentrates from the stomach and digestive diverticula of these shellfish. NoV was detected in 19.35 % (N = 155), 16.67 % (N = 114), 5.70 % (N = 158), 8.82 % (N = 136), 13.74 % (N = 131), and 16.44 % (N = 146) of oyster, mussel, scallop, razor clam, ark shell, and clam samples, respectively. The average detection rate was 13.33 % (112/840). Nucleotide sequencing of the NoV RT-PCR products demonstrated that all strains belonged to NoV genotype GII.12, except two that belonged to GI.3. More than 10² copies of the NoV genome were detected in 69 of 112 positive shellfish samples. Our results suggest that ~13 % of shellfish harbor NoV, and GII.12 NoV is the primary strain in shellfish purchased at markets in seven coastal cities in China. PMID:23412724
Ma, Li-ping; Zhao, Feng; Yao, Lin; Li, Xin-guang; Zhou, De-qing; Zhang, Rui-ling
Vibrio vulnificus, an autochthonous inhabitant of the estuarine environment, was detected in water and oysters from the Great Bay Estuary System of New Hampshire and Maine. Previously, it had not been detected north of Boston Harbor on the east coast of the United States. V. vulnificus was detected in water and shellfish samples at five out of ten sites, and only in areas that were not open to recreational shellfishing. Although samples were collected from May into December, V. vulnificus was only detected in shellfish in July and August. Water sampling began in August, and V. vulnificus persisted at one site into October. PMID:2283033
O'Neill, K R; Jones, S H; Grimes, D J
This "Two if by Sea" newsletter article highlights proceedings from a workshop in 2000 that focused on shellfish disease, particularly as it relates to the aquaculture industry. Speakers, including pathologists, growers, and industry representatives, delivered a recurring message: shellfish diseases are not only here, they are here to stay. And, most likely, they concur, additional diseases will be discovered in the future. The article discusses the following shellfish diseases: MSX (multinucleated sphere unknown), dermo (short for Dermocystidium marinus, the initial classification of the parasite that causes the disease; the parasite was later found to be of the genus Perkinsus), JOD (juvenile oyster disease), and QPX (Quahog Parasite Unknown).
We performed a neuroblastoma cell (Neuro2a) culture assay modified slightly from a method reported previously to provide a simple and sensitive evaluation of paralytic shellfish poisoning (PSP) toxicity in shellfish. The cell bioassay was just as sensitive for C-toxins as for gonyautoxins. The sensitivity of our cell bioassay was 4 times that of the current standard mouse bioassay. Using the cell bioassay, we evaluated PSP toxicity in 361 shellfish samples collected from Mikawa Bay and Ise Bay, Aichi Prefecture, Japan, from April 1999-March 2002. The results were compared with those obtained in a postcolumn derivatization liquid chromatographic analysis. PSP toxins were detected in 236/361 samples by both assays, and there was a fairly good correlation (r = 0.9001, n = 236, p < 0.001) between the results from the two assays. We applied this cell bioassay when short-necked clams in the bay turned poisonous in 2001. The chronological changes in PSP toxicity in the short-necked clams were analyzed and compared with those of the cell density of poisonous plankton (Alexandrium tamarense) occurring in the bay. The PSP toxicity in shellfish peaked 2 weeks after the cell density reached a maximum. We recommend using the cell bioassay for routine monitoring of PSP toxicity in shellfish living in natural marine environments. PMID:16417278
Hayashi, Rumiko; Saito, Hiroshi; Okumura, Masanao; Kondo, Fumio
In the present work, we aimed at determining the relationship between the hepatitis A virus (HAV) numbers in imported frozen coquina clams involved in two hepatitis outbreaks, as well as the risk for human health. Due to HAV unculturability, a standardized TaqMan real-time reverse transcription-PCR controlling the virus/nucleic acid extraction and enzyme efficiencies was employed to figure the exposure dose for clams responsible for hepatitis cases. HAV numbers were then employed to figure the risk of infection based on a dose-response model for echovirus 12. The estimated risk of infection after consumption of lightly cooked clams matched actual attack rates. Our data show that prospective monitoring of bivalve samples may fail to prevent the occurrence of outbreaks, since HAV was detected in 44% of samples directly associated with cases but was undetectable in samples that were randomly collected from the importers and belonged to the same batches. A correlation was nevertheless observed between the prevalence of hepatitis A cases in the harvesting areas and positive HAV isolation in clams, which points to the need to identify and prevent hazards rather than relying on random sampling of finished products to ensure safety. However, when evidence shows that a critical limit of viral contamination has been exceeded in the potential sources of contamination discharging into the shellfish-growing beds, quantitative virological analysis addressing quality assurance and quality control requirements should be performed with the bivalves. This work provides the first evidence of accurate HAV levels in shellfish involved in outbreaks that could be of use for risk assessment purposes.
Pinto, Rosa M.; Costafreda, M. Isabel; Bosch, Albert
The interaction between paralytic shellfish poison (PSP) and melanin from butter clam (Saxidomus giganteus) and synthetic melanin followed a pattern similar to that reported between PSP and a cation exchanger. The close relation between the anatomical dis...
R. J. Price J. S. Lee
This study determined the concentrations of major and trace elements in shellfish (oysters, clams and mussels) and conducted an assessment of the health risks due to the consumption of contaminated seafood. Samples were collected at 34 sites along Todos os Santos Bay, Brazil. The elements were determined by ICP OES and Hg by Direct Mercury Analysis. Relatively high concentrations of trace elements (As, Zn, Se and Cu) were found in seafood tissues. Potential daily intake of As, Co, Se, Zn and Cu associated to shellfish consumption suggested relevant non-carcinogenic risk for all studied locations. Copper was the element that posed the greatest non-carcinogenic risk, while Pb posed the highest carcinogenic risk. Health risks for humans were greatest from the consumption of mussels. Contaminated shellfish offer the greatest risk for children, subsistence fishers and subsistence shellfish consumers. PMID:21803378
de Souza, Manuel M; Windmöller, Cláudia C; Hatje, Vanessa
Executive Summary Historically, the Peconic Estuary's shellfish resources have supported significant fisheries for a number of species including hard clams, oysters and bay scallops. However, distribution and abundance data for the tributaries and embayments within the Peconic Es...
...2010-10-01 false Subsistence taking of fish, wildlife, and shellfish: general regulations... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE...LANDS IN ALASKA Subsistence Taking of Fish and Wildlife Â§ 100.25...
...2009-10-01 false Subsistence taking of fish, wildlife, and shellfish: general regulations... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE...LANDS IN ALASKA Subsistence Taking of Fish and Wildlife Â§ 100.25...
Contents: Summary of activities of the Gulf Coast Shellfish Sanitation Research Center; Effect of turbidity on the rate of accumulation and elimination of Poliovirus Type 1 by the eastern oyster, Crassostrea virginica; Effect of ultraviolet light on survi...
R. J. Hammerstrom W. F. Hill
This coloring book contains pictures of more than 79 fish and shellfish found on the Atlantic Coast. Captions give information on habitats, behavior, or commercial uses of the species pictured. Indexes of both common and scientific names are given. (BB)
Flick, George J.
Paralytic shellfish poisoning (PSP), a human illness caused by the ingestion of shellfish contaminated with paralytic shellfish toxins (PSTs), has been reported in Alaska for decades. These poisoning incidents have resulted in losses to local economies due to shellfish harvest closures. Thus the development of an effective biotoxin monitoring program designed specifically for the remote regions of Alaska would provide protection for public health and allow for a viable shellfish industry. The present study provides data useful for the development of an effective toxin screening protocol by comparing PST levels quantified in shellfish by many of the currently available PST detection techniques. Seven bivalve species were collected along beaches of the Aleutian Islands from June 2006 to September 2007. The concentration of PSTs was quantified and compared using five different analytical methods: the mouse bioassay, high performance liquid chromatography (HPLC), receptor-binding assay, the commercially available Jellett Rapid PSP Test strips, and an enzyme linked immunosorbent assay technique. The Association of Official Analytical Chemists (AOAC)-approved HPLC method proved to be valuable for characterizing the suite of individual PSTs in each species for research purposes, but was not considered practical for rapid toxin screening in remote Alaskan regions due to its time-consuming nature and requirement of expensive equipment and considerable expertise. In the present study, Jellett test strips were shown to be an effective tool for rapid screening, however due to the high percentage of false positives, subsequent validation via AOAC-approved methods would be required to prevent unnecessary closures. PMID:19450616
Costa, Pedro Reis; Baugh, Keri A; Wright, Bruce; Ralonde, Raymond; Nance, Shelly L; Tatarenkova, Natália; Etheridge, Stacey M; Lefebvre, Kathi A
The problem of bacterial pollution in shellfishing areas is not uncommon in the coastal regions of the United States. Bacterial contamination from man's activities can effectively reduce our natural shellfish resource areas by forcing their closure because of high potential risk of diseases being spread by shellfish harvested in these areas. Tillamook Bay, a relatively small, enclosed drainage basin of nonurban character, presents an excellent study area for observing this problem. The high population density of animals, raised on a relatively small floodplain area, represents one of the major sources of pollution in the bay. This paper summarizes the history of the agencies involved with the problem and presents the current approach to alleviate bacterial pollution in the bay without unduly penalizing other industries in the Tillamook basin. The paper also presents some of the legal aspects of reducing water pollution in shellfish harvesting areas and the jurisdiction of federal agencies in these matters. Finally, recommendations are given to reduce bacterial output by the major source categories in the basin, and criteria for bay closure to shellfish harvest are developed to protect the public from bacterially contaminated shellfish.
Crane, Stuart R.; Moore, James A.
The illness of three people due to diarrhetic shellfish poisoning (DSP) following their ingestion of recreationally harvested mussels from Sequim Bay State Park in the summer of 2011, resulted in intensified monitoring for diarrhetic shellfish toxins (DSTs) in Washington State. Rapid testing at remote sites was proposed as a means to provide early warning of DST events in order to protect human health and allow growers to test "pre-harvest" shellfish samples, thereby preventing harvest of toxic product that would later be destroyed or recalled. Tissue homogenates from several shellfish species collected from two sites in Sequim Bay, WA in the summer 2012, as well as other sites throughout Puget Sound, were analyzed using three rapid screening methods: a lateral flow antibody-based test strip (Jellett Rapid Test), an enzyme-linked immunosorbent assay (ELISA) and a protein phosphatase 2A inhibition assay (PP2A). The results were compared to the standard regulatory method of liquid chromatography coupled with tandem mass spectroscopy (LC-MS/MS). The Jellett Rapid Test for DSP gave an unacceptable number of false negatives due to incomplete extraction of DSTs using the manufacturer's recommended method while the ELISA antibody had low cross-reactivity with dinophysistoxin-1, the major toxin isomer in shellfish from the region. The PP2A test showed the greatest promise as a screening tool for Washington State shellfish harvesters. PMID:24084788
Eberhart, Bich-Thuy L; Moore, Leslie K; Harrington, Neil; Adams, Nicolaus G; Borchert, Jerry; Trainer, Vera L
Aims: This study was carried out to evaluate the presence of Shiga toxin-producing Escherichia coli (STEC) and E. coli O157:H7 in shellfish from French coastal environments. Methods and Results: Shellfish were collected in six growing areas or natural beds (B-category) and non-farming areas (D-category) from July 2002 to August 2004. PCR-detection of stx genes was performed on homogenized whole shellfish
M. Gourmelon; M. P. Montet; S. Lozach; C. Le Mennec; M. Pommepuy; L. Beutin; C. Vernozy-Rozand
Routine monitoring along the coast of the Gulf of Maine (GoM) reveals shellfish toxicity nearly every summer, but at varying times, locations, and magnitudes. The responsible toxin is known to be produced by the dinoflagellate Alexandrium fundyense, yet there is little apparent association between Alexandrium abundance and shellfish toxicity. One possibility is that toxic cells are persistent in offshore areas and variability in shellfish toxicity is caused not by changes in overall abundance, but rather by variability in transport processes. Measurements of offshore Alexandrium biomass are scarce, so we bypass cell abundance as an explanatory variable and focus instead on the relations between shellfish toxicity and concurrent metrics of GoM meteorology, hydrology, and oceanography. While this yields over two decades (1985–2005) of data representing a variety of interannual conditions, the toxicity data are gappy in spatial and temporal coverage. We address this through a combination of parametric curve fitting and hierarchical cluster analysis to reveal eight archetypical modes of seasonal toxicity timing. Groups of locations are then formed that have similar interannual patterns in these archetypes. Finally, the interannual patterns within each group are related to available environmental metrics using classification trees. Results indicate that a weak cross-shore sea surface temperature (SST) gradient in the summer is the strongest correlate of shellfish toxicity, likely by signifying a hydrological connection between offshore Alexandrium populations and near-shore shellfish beds. High cumulative downwelling wind strength early in the season is revealed as a precursor consistent with this mechanism. Although previous studies suggest that alongshore transport is important in moving Alexandrium from the eastern to western GoM, alongshore SST gradient is not an important correlate of toxicity in our study. We conclude by discussing the implications of our results for designing efficient and effective shellfish monitoring programs along the GoM coast.
Nair, Apurva; Thomas, Andrew C.; Borsuk, Mark E.
Routine monitoring along the coast of the Gulf of Maine (GoM) reveals shellfish toxicity nearly every summer, but at varying times, locations, and magnitudes. The responsible toxin is known to be produced by the dinoflagellate Alexandrium fundyense, yet there is little apparent association between Alexandrium abundance and shellfish toxicity. One possibility is that toxic cells are persistent in offshore areas and variability in shellfish toxicity is caused not by changes in overall abundance, but rather by variability in transport processes. Measurements of offshore Alexandrium biomass are scarce, so we bypass cell abundance as an explanatory variable and focus instead on the relations between shellfish toxicity and concurrent metrics of GoM meteorology, hydrology, and oceanography. While this yields over two decades (1985-2005) of data representing a variety of interannual conditions, the toxicity data are gappy in spatial and temporal coverage. We address this through a combination of parametric curve fitting and hierarchical cluster analysis to reveal eight archetypical modes of seasonal toxicity timing. Groups of locations are then formed that have similar interannual patterns in these archetypes. Finally, the interannual patterns within each group are related to available environmental metrics using classification trees. Results indicate that a weak cross-shore sea surface temperature (SST) gradient in the summer is the strongest correlate of shellfish toxicity, likely by signifying a hydrological connection between offshore Alexandrium populations and near-shore shellfish beds. High cumulative downwelling wind strength early in the season is revealed as a precursor consistent with this mechanism. Although previous studies suggest that alongshore transport is important in moving Alexandrium from the eastern to western GoM, alongshore SST gradient is not an important correlate of toxicity in our study. We conclude by discussing the implications of our results for designing efficient and effective shellfish monitoring programs along the GoM coast. PMID:23391892
Nair, Apurva; Thomas, Andrew C; Borsuk, Mark E
Of the three types of toxicity known so far in Portuguese shellfish, only diarrhetic shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) are produced by microalgae that seem to have been present in the last decades or centuries. The most important paralytic shellfish poisoning (PSP) producer, Gymnodinium catenatum, is hypothesised to have been introduced quite recently as only in 1976 PSP toxicity was detected for the first time in shellfish from Galicia, NW Iberian Peninsula. While ASP presents very short episodes of contamination, the concentration of DSP toxins in some years surpasses human safety values for much longer periods. It is traditionally stated that shellfish should be consumed in 'months with R' (September-April). A retrospective study of the maximum monthly DSP levels attained in mussels from a coastal lagoon-Ria de Aveiro-between 1994 and 2001, showed that the highest frequency of months with concentrations surpassing the safety level of 2 microg/g digestive glands were found in June-September, followed by May and October. These months correspond with the months of lowest historical average rainfall in the period 1941-1998. Oscillations in the rainfall pattern coincided with earliest (or latest) detection by HPLC of DSP toxins in mussel in the years studied. In a semi-closed lagunar environment prone to in situ growth of DSP-producer microalgae, like Dinophysis acuminata, rainfall affects river output, lowering salinity and disrupting water column stability that favours Dinophysis growth. The seasonality of DSP recurrence may be connected to the folk adage on safety of shellfish consumption, after many years of empirical observations by coastal populations of diarrhoea episodes in summertime. PMID:12565738
Vale, Paulo; Sampayo, Maria Antónia de M
Common blue mussels (Mytilus edulis), horse mussels (Modiolus modiolus), and flat oysters (Ostrea edulis) obtained from various harvesting and commercial production sites along the Norwegian coast were screened for the presence of norovirus by a real-time reverse transcription (RT)-nested PCR assay and for possible indicators of fecal contamination, i.e., for F-specific RNA bacteriophages (F-RNA phages) by plaque assay and for human adenoviruses and human circoviruses by nested PCR assay. The aims were to obtain relevant information for assessing the risk of transmission of enteric viruses by shellfish and to investigate the potential of various indicator viruses in routine screening. Noroviruses were detected in 6.8% of the samples, and the indicators were detected in 23.8% (F-RNA phages), 18.6% (adenoviruses), and 8.0% (circoviruses) of the samples. A seasonal variation was observed, with the exception of circoviruses, with more positive samples in the winter. A positive correlation was found between F-RNA phages and noroviruses. However, F-RNA phages were present in only 43% of the norovirus-positive samples. The results show that mussels from the Norwegian coast can constitute a risk of infection with enteric viruses and that routine testing of samples may be justified. Advantages and disadvantages of various options for screening are discussed.
Myrmel, M.; Berg, E. M. M.; Rimstad, E.; Grinde, B.
Changes in free amino acid content and compostion, and formation of odor components in ark shell Anadara broughtonii(Shrenk) during frozen storgae were studied in connection with storage temperature. The shucked shell-fish were frozen -40°C and stored at -10, -20 and -40°C, respectively for three months, and the shell flesh were subjected to anyalyses for free amino acids and odor components. The amount of total free amino acids in the shell flesh storedat -10°C was not as large as those in the flesh stored at -20 and -40°C. This finding implies that free amino acids in the shell flesh, when frozen-stored at a relatively high temperature, decrease in amount during storage and/or become liable to be lost along with drip during the course of thawing. As for respective amino acids, a mounts of Tau, ?-Ala, and Gly decreased markedly, while those of Met, Leu and Ile increased to some extent. In addition, it is of particular interest that ?-Ala is likely to be present in the free state. Amounts of volatile compounds such as carbonyl, nitrogenous, and sulfurous compounds have never attained the level where unpleasant or fishy odor could be percieved. However, there was a clear tendency that the higher the storage temperature, the larger amount of these compounds were presen.
Song, Dae Jin; Konagaya, Shiro; Nakamura, Koji; Iida, Haruka; Tanaka, Takeo
Filter-feeding bivalve molluscs are often contaminated by algal toxins. We have probed whether proteomic analysis of extracts from the digestive gland (DG) of mussels could be employed to identify biomarkers of contamination due to okadaic acid-group toxins. The protein extracts were obtained from 18 separate mussel samples and were analyzed by two-dimensional gel electrophoresis. When samples were divided into four different classes based on the content of OA-group toxins in the starting material, we found that two proteins varied as a function of OA contamination. By BLAST analysis, the two proteins were identified as a component of photosystem II and a subunit of NADH dehydrogenase. The analysis of peptide homologies showed that the peptide of photosystem II we detected in extracts from the DG of mussels contaminated by OA-group toxins is identical to its counterpart in Dinophysis algae, which are the producers of this group of toxins. We concluded that proteomic analysis can be used for the detection and identification of biomarkers of biotoxin contamination in shellfish, including both proteins expressed by the toxin producers and components that participate to the tissue response to the exogenous bioactive contaminant. PMID:18782583
Ronzitti, Giuseppe; Milandri, Anna; Scortichini, Giampiero; Poletti, Roberto; Rossini, Gian Paolo
Paralytic shellfish toxins (PSTs) are non-protein neurotoxins produced by saltwater dinoflagellates and freshwater cyanobacteria. The ability of Lactobacillus rhamnosus strains GG and LC-705 (in viable and non-viable forms) to remove PSTs (saxitoxin (STX), neosaxitoxin (neoSTX), gonyautoxins 2 and 3 (GTX2/3), C-toxins 1 and 2 (C1/2)) from neutral and acidic solution (pH 7.3 and 2) was examined using HPLC. Binding decreased in the order of STX ~ neoSTX > C2 > GTX3 > GTX2 > C1. Removal of STX and neoSTX (77%–97.2%) was significantly greater than removal of GTX3 and C2 (33.3%–49.7%). There were no significant differences in toxin removal capacity between viable and non-viable forms of lactobacilli, which suggested that binding rather than metabolism is the mechanism of the removal of toxins. In general, binding was not affected by the presence of other organic molecules in solution. Importantly, this is the first study to demonstrate the ability of specific probiotic lactic bacteria to remove PSTs, particularly the most toxic PST-STX, from solution. Further, these results warrant thorough screening and assessment of safe and beneficial microbes for their usefulness in the seafood and water industries and their effectiveness in vivo.
Vasama, Mari; Kumar, Himanshu; Salminen, Seppo; Haskard, Carolyn A.
Use of SNPs has been favoured due to their abundance in plant and animal genomes, accompanied by the falling cost and rising throughput capacity for detection and genotyping. Here, we present in vitro (obtained from targeted sequencing) and in silico discovery of SNPs, and the design of medium-throughput genotyping arrays for two oyster species, the Pacific oyster, Crassostrea gigas, and European flat oyster, Ostrea edulis. Two sets of 384 SNP markers were designed for two Illumina GoldenGate arrays and genotyped on more than 1000 samples for each species. In each case, oyster samples were obtained from wild and selected populations and from three-generation families segregating for traits of interest in aquaculture. The rate of successfully genotyped polymorphic SNPs was about 60% for each species. Effects of SNP origin and quality on genotyping success (Illumina functionality Score) were analysed and compared with other model and nonmodel species. Furthermore, a simulation was made based on a subset of the C. gigas SNP array with a minor allele frequency of 0.3 and typical crosses used in shellfish hatcheries. This simulation indicated that at least 150 markers were needed to perform an accurate parental assignment. Such panels might provide valuable tools to improve our understanding of the connectivity between wild (and selected) populations and could contribute to future selective breeding programmes. PMID:24447767
Lapègue, S; Harrang, E; Heurtebise, S; Flahauw, E; Donnadieu, C; Gayral, P; Ballenghien, M; Genestout, L; Barbotte, L; Mahla, R; Haffray, P; Klopp, C
Paralytic shellfish toxins (PSTs) are non-protein neurotoxins produced by saltwater dinoflagellates and freshwater cyanobacteria. The ability of Lactobacillus rhamnosus strains GG and LC-705 (in viable and non-viable forms) to remove PSTs (saxitoxin (STX), neosaxitoxin (neoSTX), gonyautoxins 2 and 3 (GTX2/3), C-toxins 1 and 2 (C1/2)) from neutral and acidic solution (pH 7.3 and 2) was examined using HPLC. Binding decreased in the order of STX ~ neoSTX > C2 > GTX3 > GTX2 > C1. Removal of STX and neoSTX (77%-97.2%) was significantly greater than removal of GTX3 and C2 (33.3%-49.7%). There were no significant differences in toxin removal capacity between viable and non-viable forms of lactobacilli, which suggested that binding rather than metabolism is the mechanism of the removal of toxins. In general, binding was not affected by the presence of other organic molecules in solution. Importantly, this is the first study to demonstrate the ability of specific probiotic lactic bacteria to remove PSTs, particularly the most toxic PST-STX, from solution. Further, these results warrant thorough screening and assessment of safe and beneficial microbes for their usefulness in the seafood and water industries and their effectiveness in vivo. PMID:25046082
Vasama, Mari; Kumar, Himanshu; Salminen, Seppo; Haskard, Carolyn A
Monitoring of amnesic shellfish poisoning (ASP) toxins in shellfish from the Catalan coast started in 2001. No ASP toxins were detected in any of the analyses performed before 2008. On 22 January 2008, domoic acid (DA) was detected in Donax trunculus (0.5 mg?kg(-1)) and confirmed by rapid resolution liquid chromatography-tandem mass spectrometry (0.6 mg?kg(-1)). A total of 974 shellfish samples were analyzed from January 2008 to December 2011, covering all the Catalan production areas and the most important marketed species. DA was detected in 23.8 % of the samples and was recorded every month in all areas and all species, except Ostrea edulis, although the percentage of samples with DA and DA content varied widely among samples. DA exceeded the regulatory level of 20 mg?kg(-1) twice: in Callista chione sampled on February 2008 and in D. trunculus sampled on April 2010. DA content in Bolinus brandaris appeared constant and close to 4.5 mg?kg(-1) for months in 2009. Mytilus galloprovincialis, Crassostrea gigas, and Ruditapes sp. presented very low concentrations of DA in the Ebro Delta bays, despite 113 alert situations according to Pseudo-nitzschia spp. abundances and the high number of shellfish samples analyzed. The origin of DA in Catalan shellfish remains unknown. PMID:23275095
Giménez Papiol, Gemma; Casanova, Alexis; Fernández-Tejedor, Margarita; de la Iglesia, Pablo; Diogène, Jorge
Purple clams, Hiatula diphos Linnaeus, are filter-feeding bivalves and maculated ivory shells. Babylonia areolata Link are carnivorous gastropods. Both shellfishes are popular seafood delicacies among the Taiwanese. Hiatula diphos were forced to contain gonyautoxins (GTXs) in this research by feeding them with cells of Alexandrium minutum Halim, a toxic dinoflagellate species responsible for the paralytic shellfish poisonings in Taiwan. The intoxicated purple clams of known toxicity and toxin composition were fed to B. areolata to observe the transmission and transformation of GTXs among this shellfish. It was found that the toxin composition in bivalve and gastropod were similar to that in dinoflagellate. Our data provide evidence for food-chain transmission of paralytic shellfish poisoning toxins, from dinoflagellate to gastropod through a filter-feeding bivalve. The transmitted GTX-I. -II. -III and -IV of A. minutum could only be found in the viscera of these shellfish. There was a notable degradation of GTX-I in the ivory shell that resulted in a decrease in toxicity while the total amount of toxins was accumulatively increasing. PMID:9637371
Chen, C Y; Chou, H N
Escherichia coli is a widely utilized indicator of the sanitary quality of bivalve molluscan shellfish sold for human consumption. However, it is now well documented that shellfish that meet the E. coli standards for human consumption may contain human enteric viruses that cause gastroenteritis and hepatitis. In this study we investigated using F-specific RNA bacteriophage (FRNA bacteriophage) to indicate the
WILLIAM J. DORE; KATHLEEN HENSHILWOOD; DAVID N. LEES
Shellfish aquaculture is an expanding industry in coastal British Columbia, Canada, and occurs in important wintering areas for surf scoters (Melanitta perspicillata) and white-winged scoters (M. fusca). We quantified habitat use by scoters in relation to natural environmental attributes and habitat modifications associated with shellfish aquaculture. We found that, despite the extensive clam and oyster farming in our study area,
DEBORAH L. LACROIX
In recent years the marine environment has been seriously damaged by the presence of several toxic phytoplanktonic species, such as dinoflagellates and other toxic algae, which contaminate shellfish and other marine products. Amnesic and paralytic shellfish toxins are examples of these contaminants. The search for sensitive methodologies for the analysis of such compounds is one of the aims of researchers
N Piñeiro; J. M Leão; A Gago Mart??nez; J. A Rodr??guez Vázquez
Aquatic toxins are responsible for a number of acute and chronic diseases in humans. Okadaic acid (OA) and other dinoflagellate derived polyketide toxins pose serious health risks on a global scale. Ingestion of OA contaminated shellfish causes diarrheic shellfish poisoning (DSP). Some evidence also suggests tumor promotion in the liver by OA. Microcystin-LR (MC-LR) is produced by cyanobacteria and is
This paper describes the implementation of a wireless sensor network for the temperature monitoring of shellfish catches over the internet. Temperature loggers in shellfish boxes transmit data through radio frequency to a base station. The information is transferred to a server via the GSM network, where it is processed and uploaded to a database. The web-based interface allows configuration of
S. Palaniswamy; S. Balamurugan; A. Kumar
The phospholipid molecular species of freshwater (pangasius, Nile perch, trout), marine fish fillets (horse mackerel, European hake, common sole, European anchovy, European pilchard, Atlantic mackerel) and the edible muscle foot of bivalves (clam, mussel, oyster) commonly available in the Italian market during spring and summer were characterized by means of normal-phase high performance liquid chromatography coupled online with positive electrospray ionization ion-trap tandem mass spectrometry. From principal component analysis (PCA), it was observed that the total fatty acid profile was not suitable to differentiate among the shellfish genera. The fatty acid molecular combinations of phosphatidylcholine, the main phospholipid class, as well as phosphatidylinositol and phosphatidylethanolamine allowed for the differentiation of shellfish from the bony fishes. Phosphatidylserine and phosphatidylethanolamine plasmalogen profile allowed for the discrimination of each bony fish or shellfish genus since PS and pPE classes included a large number of fatty acid combinations that were specific for a fish genus or group. PMID:22369175
Boselli, Emanuele; Pacetti, Deborah; Lucci, Paolo; Frega, Natale G
Natural phytoplankton blooms of the dinoflagellate Alexandrium minutum, milkfish (Chanos chanos) exposed to natural blooms, sediment and mangrove crab (Scylla serrata) were analysed for paralytic shellfish poisoning toxins by high-performance liquid chromatography. The toxin profiles of milkfish and mangrove crab were similar to that of A. minutum collected from blooming fishponds. In a laboratory A. minutum-blooming environment, the stomach and intestine of milkfish accumulated paralytic shellfish poisoning toxins during the exposure period. The non-visceral tissues were non-toxic. However, milkfish lost their entire body burden of toxin on the first day of transferring to a toxic algae-free environment. The result shows that milkfish concentrate paralytic shellfish poisoning toxins in digestive organs and did not retain toxins. PMID:12881129
Chou, H-N; Chung, Y-C; Cho, W-C; Chen, C-Y
An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.
Cao, Jijuan; Zheng, Jiang; Yu, Bing; Wang, Qiuyan; Xu, Junyi; Li, Aifeng
Aquaculture is emerging as one of the most viable and promising enterprises for keeping pace with the surging need for animal protein, providing nutritional and food security to humans, particularly those residing in regions where livestock is relatively scarce. With every step toward intensification of aquaculture practices, there is an increase in the stress level in the animal as well as the environment. Hence, disease outbreak is being increasingly recognized as one of the most important constraints to aquaculture production in many countries, including India. Conventionally, the disease control in aquaculture has relied on the use of chemical compounds and antibiotics. The development of non-antibiotic and environmentally friendly agents is one of the key factors for health management in aquaculture. Consequently, with the emerging need for environmentally friendly aquaculture, the use of alternatives to antibiotic growth promoters in fish nutrition is now widely accepted. In recent years, probiotics have taken center stage and are being used as an unconventional approach that has numerous beneficial effects in fish and shellfish culture: improved activity of gastrointestinal microbiota and enhanced immune status, disease resistance, survival, feed utilization and growth performance. As natural products, probiotics have much potential to increase the efficiency and sustainability of aquaculture production. Therefore, comprehensive research to fully characterize the intestinal microbiota of prominent fish species, mechanisms of action of probiotics and their effects on the intestinal ecosystem, immunity, fish health and performance is reasonable. This review highlights the classifications and applications of probiotics in aquaculture. The review also summarizes the advancement and research highlights of the probiotic status and mode of action, which are of great significance from an ecofriendly, sustainable, intensive aquaculture point of view. PMID:24419543
C De, Bidhan; Meena, D K; Behera, B K; Das, Pronob; Das Mohapatra, P K; Sharma, A P
The illness of three people in 2011 after their ingestion of mussels collected from Sequim Bay State Park, Washington State, USA, demonstrated the need to monitor diarrhetic shellfish toxins (DSTs) in Washington State for the protection of human health. Following these cases of diarrhetic shellfish poisoning, monitoring for DSTs in Washington State became formalized in 2012, guided by routine monitoring of Dinophysis species by the SoundToxins program in Puget Sound and the Olympic Region Harmful Algal Bloom (ORHAB) partnership on the outer Washington State coast. Here we show that the DSTs at concentrations above the guidance level of 16 ?g okadaic acid (OA) + dinophysistoxins (DTXs)/100 g shellfish tissue were widespread in sentinel mussels throughout Puget Sound in summer 2012 and included harvest closures of California mussel, varnish clam, manila clam and Pacific oyster. Concentrations of toxins in Pacific oyster and manila clam were often at least half those measured in blue mussels at the same site. The primary toxin isomer in shellfish and plankton samples was dinophysistoxin-1 (DTX-1) with D. acuminata as the primary Dinophysis species. Other lipophilic toxins in shellfish were pectenotoxin-2 (PTX-2) and yessotoxin (YTX) with azaspiracid-2 (AZA-2) also measured in phytoplankton samples. Okadaic acid, azaspiracid-1 (AZA-1) and azaspiracid-3 (AZA-3) were all below the levels of detection by liquid chromatography tandem mass spectrometry (LC-MS/MS). A shellfish closure at Ruby Beach, Washington, was the first ever noted on the Washington State Pacific coast due to DSTs. The greater than average Fraser River flow during the summers of 2011 and 2012 may have provided an environment conducive to dinoflagellates and played a role in the prevalence of toxigenic Dinophysis in Puget Sound.
Trainer, Vera L.; Moore, Leslie; Bill, Brian D.; Adams, Nicolaus G.; Harrington, Neil; Borchert, Jerry; da Silva, Denis A. M.; Eberhart, Bich-Thuy L.
Ciguatera fish poisoning and neurotoxic shellfish poisoning are distinct clinical entities characterized by gastrointestinal and neurological disturbances, following the consumption of certain reef fish and shellfish containing toxic polyether compounds sporadically present in certain toxic marine dinoflagellates. The biotransformation and bioaccumulation of gambierol and brevetoxin, and their congeners, are believed to be involved in the pathogenesis of these "food-chain diseases", for which no effective treatments are available. Here, we describe for the first time the potent effect of gambierol and brevetoxin on TRPV1 channels, a key player in thermal and pain sensation. Our findings may lead to promising new therapeutic interventions. PMID:17659256
Cuypers, Eva; Yanagihara, Angel; Rainier, Jon D; Tytgat, Jan
Objective is to investigate the feasibility of employing food irradiation technology to reduce or eliminate the threat of viral diseases contracted as a result of consumption of raw or inadequately cooked shellfish. Several recently published studies warn of the health risks associated with eating of raw shellfish - particularly American oysters, Crassostrea virginica, and the hardshelled clam, Mercenaria mercenaria. This study addresses the possibility of reducing the incidence of molluscanborne diseases through the application of /sup 60/Co gamma irradiation processing to effect the inactivation of pathogenic viruses in live, raw shwllfish. Dosimetry, D/sub 10/ doses, and organoleptic effects were studied.
Beghian, L.; Melnick, J.
In 2011, a Diarrhetic Shellfish Poisoning (DSP) outbreak occurred in British Columbia (BC), Canada that was associated with cooked mussel consumption. This is the first reported DSP outbreak in BC. Investigation of ill individuals, traceback of product and laboratory testing for toxins were used in this investigation. Sixty-two illnesses were reported. Public health and food safety investigation identified a common food source and harvest area. Public health and regulatory agencies took actions to recall product and notify the public. Shellfish monitoring program changes were implemented after the outbreak. Improved response and understanding of toxin production will improve management of future DSP outbreaks.
Taylor, Marsha; McIntyre, Lorraine; Ritson, Mark; Stone, Jason; Bronson, Roni; Bitzikos, Olga; Rourke, Wade; Galanis, Eleni
AIM: To assess current practice of United Kingdom cardiologists with respect to patients with reported shellfish/iodine allergy, and in particular the use of iodinated contrast for elective coronary angiography. Moreover we have reviewed the current evidence-base and guidelines available in this area. METHODS: A questionnaire survey was send to 500 senior United Kingdom cardiologists (almost 50% cardiologists registered with British Cardiovascular Society) using email and first 100 responses used to analyze practise. We involved cardiologists performing coronary angiograms routinely both at secondary and tertiary centres. Three specific questions relating to allergy were asked: (1) History of shellfish/iodine allergy in pre-angiography assessment; (2) Treatments offered for shellfish/iodine allergy individuals; and (3) Any specific treatment protocol for shellfish/iodine allergy cases. We aimed to establish routine practice in United Kingdom for patients undergoing elective coronary angiography. We also performed comprehensive PubMed search for the available evidence of relationship between shellfish/iodine allergy and contrast media. RESULTS: A total of 100 responses were received, representing 20% of all United Kingdom cardiologists. Ninety-three replies were received from consultant cardiologists, 4 from non-consultant grades and 3 from cardiology specialist nurses. Amongst the respondents, 66% routinely asked about a previous history of shellfish/iodine allergy. Fifty-six percent would pre-treat these patients with steroids and anti-histamines. The other 44% do nothing, or do nonspecific testing based on their personal experience as following: (1) Skin test with 1 mL of subcutaneous contrast before intravenous contrast; (2) Test dose 2 mL contrast before coronary injection; (3) Close observation for shellfish allergy patients; and (4) Minimal evidence that the steroid and anti-histamine regime is effective but it makes us feel better. CONCLUSION: There is no evidence that allergy to shellfish alters the risk of reaction to intravenous contrast more than any other allergy and asking about such allergies in pre-angiogram assessment will not provide any additional information except propagating the myth.
Baig, Mudassar; Farag, Ahmad; Sajid, Jamal; Potluri, Rahul; Irwin, R Bruce; Khalid, Hafiz Mohammed Idrees
In 2011, a Diarrhetic Shellfish Poisoning (DSP) outbreak occurred in British Columbia (BC), Canada that was associated with cooked mussel consumption. This is the first reported DSP outbreak in BC. Investigation of ill individuals, traceback of product and laboratory testing for toxins were used in this investigation. Sixty-two illnesses were reported. Public health and food safety investigation identified a common food source and harvest area. Public health and regulatory agencies took actions to recall product and notify the public. Shellfish monitoring program changes were implemented after the outbreak. Improved response and understanding of toxin production will improve management of future DSP outbreaks. PMID:23697950
Taylor, Marsha; McIntyre, Lorraine; Ritson, Mark; Stone, Jason; Bronson, Roni; Bitzikos, Olga; Rourke, Wade; Galanis, Eleni
The Fishery Biology Program of the Northeast Fisheries Science Center provides this resource to assist with age determination of fish and shellfish species from the Northwest Atlantic. Released in its current form in 1997, this resource represents the fruit of many years of accumulated expertise. Techniques for aging sixteen species of fish or shellfish are given here, based on growth marks in scales, otoliths, and shells. From Atlantic Butterfish through Yellowtail Flounder, these detailed descriptions and black-and-white images should be of valuable assistance to researchers.
Almeida, F. P.; Dery, L. M.; Penttila, J.; Sheehan, T. F.
SUMMARY The North Sea supports a fishing and offshore oil and gas production industry, some of the most heavily exploited resources from the seas around the United Kingdom. This report addresses the concern that some commercially exploited fish stocks could potentially be contaminated by hydrocarbons from the offshore oil industry. Fish and shellfish caught in July\\/August 2002 in the East
A D McIntosh; A E Craig
A single-laboratory validation is reported for an LC/MS/MS quantification of six brevetoxins in four matrixes (Greenshell mussel, eastern oyster, hard clam, and Pacific oyster). Recovery and precision data were collected from seven analytical batches using shellfish flesh at 0.05 mg/kg. Method recoveries and within-laboratory reproducibility ranged from 73 to 112%, with an RSD between 14 and 18% for brevetoxin-3, brevetoxin B5, brevetoxin B2, and S-desoxy brevetoxin B2. The recovery and within-laboratory reproducibility for brevetoxin-2 was 61%, with an RSD of 27%. Brevetoxin B1 gave an RSD of 12%, but no reference material was available and this toxin was only recorded in a hard clam sample naturally contaminated with brevetoxins. One naturally contaminated sample of each shellfish matrix, with brevetoxin levels ranging from 0.012 to 9.9 mg/kg, was tested in multiple batches, and the RSDs were similar to those for fortified samples at 0.05 mg/kg. Comparisons with limited data for the neurotoxic shellfish poisoning mouse bioassay for four naturally contaminated shellfish samples showed that the regulatory action limit of 0.8 mg/kg is conservative with respect to the bioassay regulatory limit of 20 mouse units/100 g. PMID:22970578
McNabb, Paul S; Selwood, Andrew I; Van Ginkel, Roel; Boundy, Michael; Holland, Patrick T
BACKGROUND: Recommendations about risks and benefits of seafood intake during pregnancy have been published in the last decade, but the specific health effects of the different categories of seafood remain unknown. Fish and shellfish may differ according to their fatty acid content and their concentration of chemical pollutants and toxins. Not taking these particularities into account may result in underestimating
Laurence Guldner; Christine Monfort; Florence Rouget; Ronan Garlantezec; Sylvaine Cordier
In Japan, seed production techniques have been exploited for about 80 species of marine fish and shellfish. However, mass mortalities due to infectious and non-infectious diseases have often occurred in larvae and juveniles reared in marine hatcheries. Among these problems the viral and bacterial diseases with their control measures are reviewed in this paper. Since around the middle of the
Hepatitis A virus (HAV) represents a significant public health problem due to its high persistence in the environment and its transmission through contaminated water and food. Bivalve shellfish are filter feeders that can bioaccumulate human pathogens found in contaminated waters, their consumption being a potential cause of hepatitis A outbreaks. In this work, cultured and wild bivalve shellfish from the Ría de Vigo (Galicia, NW Spain) were analysed for the presence and genotyping of HAV. A total of 160 shellfish samples were collected between March 2004 and December 2006, including 68 samples from cultured mussels (Mytilus galloprovincialis), 30 from wild clams (Rupitapes decussatus), 31 from wild cockles (Cerastoderma edule) and 31 from wild mussel. HAV detection, carried out by quantitative RT-PCR, was positive for 29 (42.6%) cultured and 40 (43.5%) wild samples, with levels ranging from 3.1 x 10(2) and 1.4 x 10(10) RNA copies/g of shellfish digestive tissue. The phylogenetic analysis of VP1-P2A and VP3-VP1 regions, separately or as concatenated sequences, revealed that all HAV strains analysed belong to subgenotype IB. These results indicate a high prevalence of this subgenotype in the area studied. PMID:20057087
Manso, C F; Polo, D; Vilariño, M L; Romalde, J L
The main diarrhetic shellfish toxins, okadaic acid (OA) and dinophysistoxin-1, 2 (DTX-2, 2) were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) as pyrenacyl esters in clams (Ruditapes decussatus) collected in Tunis north lagoon from January 2007 to June 2008. Sample analyses by LC-MS/MS displayed OA and related congeners (DTX-2, 2) with a highest detected level of 21 ?g OA eq/kg shellfish meat for the samples of January 2007. Nevertheless, all samples were MBA negative. During the study period, potentially toxic dinoflagellate Dinophysis sacculus was recorded all year, blooming at different times. Highest concentrations were recorded during January 2007 with 4.6?×?10(4) cells per liter and 4.10(4) cells per liter in the northern and southern districts, respectively. Results show that there is no significant correlation between D. sacculus densities in water column and diarrhetic shellfish poisoning (DSP) toxins concentrations unregistered in clams. These data reveal that DSP toxicity in clams of Tunis north lagoon is low according to European regulatory limit (160 ?g OA eq/kg shellfish meat). However, a potential threat, in this area, is represented by DSP toxic species as D. sacculus and provides grounds for widen and reinforcing sanitary control of the phycotoxin measures in the region. PMID:21922181
Armi, Zina; Turki, Souad; Trabelsi, Elbahri; Ceredi, Alfiero; Riccardi, Elena; Milandri, Anna
A research element of the European Union (EU) sixth Framework project BioCop focused on the development of a surface plasmon resonance (SPR) biosensor assay for the detection of paralytic shellfish poisoning (PSP) toxins in shellfish as an alternative to the increasingly ethically unacceptable mouse bioassay. A biosensor assay was developed using both a saxitoxin binding protein and chip surface in tandem with a highly efficient simple extraction procedure. The present report describes the single laboratory validation of this immunological screening method, for this complex group of toxins with differing toxicities, according to the European Decision 2002/657/EC in conjunction with IUPAC and AOAC single laboratory validation guidelines. The different performance characteristics (detection capability CCbeta, specificity/selectivity, repeatability, reproducibility, stability, and applicability) were determined in relation to the EU regulatory limit of 800 microg of saxitoxin equivalents (STX eq) per kg of shellfish meat. The detection capability CCbeta was calculated to be 120 microg/kg. Intra-assay repeatability was found to be between 2.5 and 12.3% and interassay reproducibility was between 6.1 and 15.2% for different shellfish matrices. Natural samples were also evaluated and the resultant data displayed overall agreements of 96 and 92% with that of the existing AOAC approved methods of mouse bioassay (MBA) and high performance liquid chromatography (HPLC), respectively. PMID:20232817
Campbell, Katrina; Haughey, Simon A; van den Top, Hester; van Egmond, Hans; Vilariño, Natalia; Botana, Luis M; Elliott, Christopher T
The current method for Paralytic Shellfish Poisoning (PSP) testing in shellfish is based on the mouse bioassay (MBA), which involves injecting shellfish extract into a conscious mouse, and then converting its time to death into PSP toxicity using Sommer's table. To improve animal welfare, the present study investigated the use of anaesthesia. A saxitoxin (STX) calibration study was conducted where known amounts of STX were injected into both unanaesthetised and anaesthetised mice. Death time was approximately doubled when mice were anaesthetised. Both unanaesthetised and anaesthetised animals showed a linear relationship between the inverse death time and log(STX). Based on these data, new calibration curves were developed. This study revealed that the current method employing Sommer's table underestimates toxicity by up to 50% for higher toxin levels. Subsequently, shellfish samples were tested on both unanaesthetised and anaesthetised mice. Using the new calibration curves, the numbers of samples exceeding the field closure limit were similar for unanaesthetised and anaesthetised mice, and were nearly two-fold higher than those obtained with the current method. The studies showed that the bioassay gives variable results for both unanaesthetised and anaesthetised animals. Anaesthesia forms a viable and more ethical alternative to the current bioassay, at least in the short term. A practical summary on how to conduct this method is given. PMID:16427671
Holtrop, G; Petrie, J; McElhiney, J; Dennison, N
During the summer of 2002 there were several episodes of human intoxication after consumption of brown crabs ( Cancer pagurus ) caught along the Norwegian south coast. The toxic agent was one of the well-known phycotoxins, the diarrhoetic shellfish poisoning (DSP) complex, routinely found in blue mussels ( Mytilus edulis ). This toxin has not previously been documented in brown
Tonje Castberg; Trine Torgersen; John Aasen; Tore Aune; Lars-Johan Naustvoll
In 2002 several hundred people were taken ill after eating self-harvested brown crabs (Cancer pagurus) in the southern part of Norway. The symptoms were similar to diarrhetic shellfish poisoning (DSP) although with a somewhat delayed onset. This happened at the same time as an unusual early bloom of Dinophysis acuta had lead to high amounts of DSP toxins in blue
Trine Torgersen; John Aasen; Tore Aune
Summary A fluorescent zebrafish was the first genetically engineered animal to be marketed, and biotechnologists are developing many transgenic fish and shellfish. Biosafety science is not sufficiently advanced to be able to draw scientifically reliable and broadly trusted conclusions about the environmental effects of these animals. The science is best developed for identifying hazards posed by environmental spread of a
A. R. Kapuscinski
The aim of this study was to evaluate the importance of biotransformation of paralytic shellfish toxins during the detoxification process in contaminated oysters. Mathematical models based upon the detoxification patterns of digestive gland and other tissues were developed. It was demonstrated that biotransformations do not seem to play an important role in digestive gland or other tissue detoxification kinetics with
Marielle Guéguen; Régis Baron; Michèle Bardouil; Philippe Truquet; Hansy Haberkorn; Patrick Lassus; Laurent Barillé; Zouher Amzil
Saxitoxin (STX) was the first known and most studied toxic component of paralytic shellfish poisoning (PSP). This toxin blocks neuronal transmission by binding to the voltage-gated Na+ channel. Although the toxin's mechanism of action is well known at the molecular level, there are still many unresolved questions about its pharmacokinetics and the PSP intoxication syndrome in mammals. Some of these
Dar??o Andrinolo; Luis F Michea; Néstor Lagos
Purple clams (Hiatula rostrata Lighttoot) accumulated paralytic shellfish poisoning (PSP) toxins produced by a toxic strain of the dinoflagellate Alexandrium minutum Halim for subsequent study of toxin distribution during depuration (detoxification by a nontoxic microalgal diet or starvation). The results confirm the data in the literature concerning the high toxicity of the digestive gland, and the depuration efficiency between feed
Chih Yu Chen; Hong Nong Chou
To investigate the potential for differential accumulation of paralytic shellfish toxins (PSTs) in various tissues of the akoya pearl oyster, Pinctada imbricata, two feeding trials were carried out using the PST-producing dinoflagellate, Alexandrium minutum. When fed with A. minutum at concentrations between 100 and 1300 cellsml?1, the maximum clearance by P. imbricata was shown to occur at a density of
Shauna A. Murray; Wayne A. O'Connor; Alfonsus Alvin; Troco K. Mihali; John Kalaitzis; Brett A. Neilan
Plankton surveys, between 2001 and 2005 along the Russian Caucasian Black Sea Coast, revealed Dinophysis rotundata, D. caudata and Prorocentrum lima as the most ubiquitous of the known dinoflagellates associated with diarrhetic shellfish poisoning (DSP). Dinophysis spp. were first observed during the spring phytoplankton succession and persist throughout the late summer phytoplankton peak. The highest total concentration, 3000cells\\/L, of D.
Steve L. Morton; Alexander Vershinin; Laurinda L. Smith; Tod A. Leighfield; Sergey Pankov; Michael A. Quilliam
A computationally designed polymer (CDP), based on the functional monomer ethylene glycol methacrylate phosphate (EGMP), with a reported high specific affinity for the neurotoxic paralytic shellfish poisoning (PSP) toxins saxitoxin (STX) and neosaxitoxin (neoSTX) was evaluated with a view to it being used in a solid phase adsorption toxin tracking (SPATT) system for deployment in the marine environment. In addition,
Paula Rodríguez; Amparo Alfonso; Elizabeth Turrell; Jean-Pierre Lacaze; Luis M. Botana
This report examines the classification of shellfish growing waters in the Gulf of Mexico as an indicator of bacterial water quality. Information presented includes the status of classified waters, sources of pollution affecting waters that are not classified as approved, and trends in classification between 1971 and 1985. Data were collected by site visits to the five Gulf states, interviews
Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prev- alent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville
W. W. CARMICHAEL; W. R. EVANS; Q. Q. YIN; P. BELL; E. MOCZYDLOWSKI
This is the first report of sheep mortalities associated with paralytic shellfish poisons (PSPs) from the cyanobacterium Anabaena circinalis Rabenhorst. Fourteen sheep died within 150 m of a farm dam containing a dense bloom of A. circinalis. Extracts from both the cyanobacterium and small intestine from a dead ewe were analysed by high-performance liquid chromatography (HPLC) and found to contain
Andrew P Negri; Gary J Jones; Michael Hindmarsh
This is a critical review of the global distribution, sources of variation in toxicity, anatomical partitioning, metabolism, and detoxification kinetics of paralytic shellfish poisoning (PSP) toxins (carbamate, TV-sulfocarbamoyl, and decarbamoyl saxitoxin derivatives) in bivalve molluscs. Marked interspecific differences in toxin accumulation are related to differences in toxin sensitivity, determined from neurological, physiological, and behavioral responses. Toxicity also varies considerably with
V. Monica Bricelj; Sandra E. Shumway
Most marine molluscan bivalve shellfish feed on suspended phytoplankton which are trapped from water pumped across the gills by ciliary action. Pathogenic microorganisms in the water may be filtered by the gills during feeding, and become concentrated in the digestive glands/tract. If these pathogens are not excreted or inactivated by the shellfish, or in subsequent preparatory processes, they may be ingested by consumers, the shellfish thereby acting as vehicles of infection. The protozoan parasites Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii have the potential to be transmitted in this way, and here we review the accumulating knowledge on the occurrence and survival of the transmission stages of these parasites in shellfish, whilst also emphasising the considerable gaps in our knowledge. Relevant information is particularly lacking for T. gondii, which, in comparison with Cryptosporidium spp. and G. duodenalis, has been relatively under-researched in this context. Although it seems evident that these shellfish can accumulate and concentrate all three of these parasites from the surrounding water, whether Giardia cysts remain viable and infectious is unknown, and some evidence suggests that they may be inactivated by the shellfish. Although both Toxoplasma and Cryptosporidium apparently retain their infectivity for prolonged periods in shellfish, the actual public health threat posed by these parasites via these shellfish is unclear, largely because there is minimal evidence of infection transmission. Reasons for this apparent lack of infection transmission are discussed and it is recommended that the potential for transmission via shellfish consumption is recognised by those concerned with investigating transmission of these infections. PMID:17928081
Robertson, L J
Matrix reference materials are an important requirement for the assessment of method performance characteristics and for routine quality control. In the field of marine toxin testing where biological assays have been used and where modern analytical testing methods are now becoming available, this requirement has become an urgent one. Various approaches are utilised for preparation of such materials in the absence of available naturally occurring toxic shellfish samples. Toxin-free shellfish may be artificially fortified through the addition of cultured toxic phytoplankton or shellfish may be incurred through natural feeding on toxic algae in a laboratory environment. Both of these approaches may be potentially affected by issues relating to the degradation or transformation of toxin analytes, so studies were conducted to assess these effects within our laboratory. A range of PSP-toxic shellfish tissues were prepared using the two approaches, in both Pacific oyster (Crassostrea gigas) and king scallops (Pecten maximus). Additionally, sub-samples of incurred Pacific oyster tissue were further treated, through addition of artificial chemical stabilisers and gamma irradiation. Two separate month-long stability trials were conducted at +4 °C on each material. Results highlighted clear evidence for improved stability of materials following shellfish feeding experiments in comparison with the tissues which had been spiked with plankton. In addition, there were clear differences in stability of toxins between the two shellfish species studied. There was evidence for good stability of C1&2 toxins in both the incurred tissues and improved stability of some toxins in tissues which had been subjected to either gamma irradiation or treatment with chemical additives. The results therefore highlighted the benefits of conducting shellfish feeding if suitable stable reference materials are to be prepared containing a full range of PSP toxin analytes. The study also highlighted the benefits of post-production treatment to prolong the stability of the materials. Work is ongoing to assess the full characteristics of candidate reference materials prepared with these approaches with the aim of producing a homogenous and stable PSP reference material in Pacific oysters. PMID:22921579
Turner, Andrew D; Lewis, Adam M; Hatfield, Robert G; Galloway, Angus W; Higman, Wendy A
Background Recommendations about risks and benefits of seafood intake during pregnancy have been published in the last decade, but the specific health effects of the different categories of seafood remain unknown. Fish and shellfish may differ according to their fatty acid content and their concentration of chemical pollutants and toxins. Not taking these particularities into account may result in underestimating of both the positive and negative effects of seafood on birth outcomes and partly explains inconsistent results on the subject. Methods In the PELAGIE cohort study, including 2398 pregnant women from Brittany, we fit multiple linear and logistic regression models to examine associations of fish (salt-water fish only) and shellfish intake before pregnancy with length of gestation, birthweight, and risks of preterm births, low birthweight or small-for-gestational-age (SGA) babies. Results When fish and shellfish consumptions were considered simultaneously, we observed a decrease in the risk of SGA birth with increasing frequency of fish intake: OR = 0.57 (95%CI: 0.31 to 1.05) for women eating fish twice a week or more compared with those eating it less than once a month. The risk of SGA birth was significantly higher among women eating shellfish twice a week or more than among those eating it less than once a month: OR = 2.14 (95%CI: 1.13 to 4.07). Each additional monthly meal including fish was significantly related to an increase in gestational length of 0.02 week (95%CI: 0.002 to 0.035). No association was observed with birthweight or preterm birth. Conclusion These results suggest that different categories of seafood may be differently associated with birth outcomes, fish consumption with increased length of gestation and shellfish consumption with decreased fetal growth.
Guldner, Laurence; Monfort, Christine; Rouget, Florence; Garlantezec, Ronan; Cordier, Sylvaine
Waste shellfish shell stacking with a significant odor and toxicity which are hazardous to human constitutes a serious environmental hazard. For utilization of waste shellfish shell resource, granule of shellfish shell (SS) was prepared from waste shellfish shell by removing cuticle, crushing, grinding and shearing emulsification and was introduced as a filler to reinforce polypropylene (PP). The mechanical behavior of PP/SS composite shows a higher yield strain, yield strength, tensile strength and elongation at break than traditional commercial calcium carbonate (CC) filled PP. Yield strength of PP/SS composite with 2% SS is improved by 11.1% due to the formation of ?-crystalline PP phase. Using waste SS for producing bio-filler for filling PP is an effective and prospective measure to deal with waste SS, which is valuable for industrial production and practical application as fillers for reinforcing polymers. PMID:22476096
Li, Hai-Yan; Tan, Ye-Qiang; Zhang, Lu; Zhang, Yun-Xiang; Song, Yi-Hu; Ye, Ying; Xia, Mei-Sheng
A new brevetoxin B analog, brevetoxin B4 (BTXB4), was isolated as the major toxin in greenshell mussels, Perna canaliculus, collected in New Zealand at the time of the neurotoxic shellfish poisoning incident. The new analog accounted for nearly two-thirds of the mouse lethality of the shellfish and was determined to be a mixture of N-myristoyl-BTXB2 and N-palmitoyl-BTXB2. PMID:10495465
Morohashi, A; Satake, M; Naoki, H; Kaspar, H F; Oshima, Y; Yasumoto, T
The Baynes Sound area accounts for a large proportion of shellfish aquaculture production in British Columbia. In response to non-industry concerns regarding impacts from this inter-tidal farming on bird populations in Baynes Sound, a quantitative inventory of aquaculture infrastructure, more specifically clam netting, was done.Low-level high-resolution air photos were taken during periods of extreme low tides, for all intertidal shellfish
Barron Carswell; Sean Cheesman; Josh Anderson
Headspace solid phase micro-extraction (HS-SPME) has been applied as a sampling technique for the determination of petroleum contamination in shellfish using capillary gas chromatography-mass spectrometry (GC-MS). A poly(dimethylsiloxane) fused silica fibre (100 µm thickness) was found to be satisfactory for the extraction of a range of aliphatic hydrocarbons (HCs) from homogenised shellfish tissues. The SPME conditions, including temperature, salt content,
Mary A. Stack; Sharon O'Connell; Kevin J. James
In the context of the French Phytoplankton and Phycotoxins Monitoring Network (REPHY) programme, shellfish samples were harvested from different locations where harmful algae blooms were known to have occurred. For all shellfish samples found positive by the mouse bioassay for diarrhetic shellfish poisoning (DSP) toxins, liquid chromatography (LC) coupled with mass spectrometry (MS) was used to search for the following lipophilic toxins: okadaic acid (OA), dinophysistoxins (DTXs), pectenotoxins (PTXs), azaspiracids (AZAs), yessotoxins (YTXs), spirolides (SPXs) and gymnodimines (GYMs). In order to investigate the presence of acyl-OAs and/or acyl-DTX-1,-2 (DTX-3), alkaline hydrolysis was performed on all samples, and LC/MS analyses were carried out on the samples before and after hydrolysis. The results revealed different lipophilic toxin profiles as a function of the shellfish sampling location. The primary finding was that all of the samples contained OA and acyl-OA. In addition, other lipophilic toxins were found in shellfish samples: DTX-2, acyl-DTX-2 and SPXs (SPX-A, SPX-desMeC) on the Atlantic coast (Southern Brittany, Arcachon), and pectenotoxins (PTX-2, PTX-2-seco-acid and 7-epi-PTX-2-seco-acid) on the Mediterranean coast (Thau lagoon, the island of Corsica). This paper reports on the first detection of PTX-2, SPX-A and their derivatives in French shellfish.
Amzil, Zouher; Sibat, Manoella; Royer, Florence; Masson, Nadine; Abadie, Eric
One hundred eighty-four serologically confirmed cases of hepatitis A were reported in eastern Spain in 1999. A matched case-control study implicated imported coquina clams complying with European Union shellfish standards as the source of infection; this implication was confirmed by the detection by reverse transcription-PCR of hepatitis A virus (HAV) RNA in shellfish samples. In spite of the recognized low variability of HAV, genetic characterization of the complete capsid region of virus isolates from patient serum samples revealed the existence of both synonymous and nonsynonymous variants. Two antigenic variants were detected, one in a discontinuous epitope defined by monoclonal antibody K3-4C8 and a second in a linear VP1 epitope of the virus. In spite of these antigenic variants, all isolates were assigned to genotype IB, providing further evidence that the outbreak originated from a common source, although multiple strains were likely to be involved. PMID:12409389
Sánchez, Glòria; Pintó, Rosa M; Vanaclocha, Hermelinda; Bosch, Albert
Heavy metals are dangerous to aquatic organisms and it can be bioaccumulated in the food chain leading to diseases in human. Cumulative effects of metals or chronic poisoning may occur as a result of long-term exposure even to low concentrations. The accumulation of heavy metals varies depending upon the species, environmental conditions, and inhibitory processes. Concentrations of zinc, copper, lead, and cadmium were determined in finfish and shellfish species in the Gangetic delta using a PerkinElmer Sciex ELAN 5000 ICP mass spectrometer and expressed as milligrams per kilogram of dry weight. In finfish and shellfish species the concentrations of Zn, Cu, Pb, and Cd were comparatively higher at stations 1 and 2 than the permissible level of WHO. The concentration of metals exhibited significant spatial variation and followed the order station 1 > station 2 > station 3 > station 4, which may be related to different degree of contamination in different location. The metal accumulation exhibited species specificity. PMID:21660552
Mitra, Abhijit; Chowdhury, Ranju; Banerjee, Kakoli
The current testing for paralytic shellfish poisoning (PSP) in shellfish is based on the mouse bioassay (MBA). To alleviate animal welfare concerns, we evaluated the utility of using sublethal indicators of toxicity as an alternative to measuring time to death. Live mice were injected with a PSP congener and the changes in neurotransmitter levels were measured 60, 90, and 120 min after injection. Acetylcholine (ACh) was the most sensitive marker for PSP toxicity. The changes in neurotransmitter levels were most pronounced in the blood. Thus, measurement of Ach levels in the blood may serve as a sensitive predictor for PSP that would not require sacrifice of the mice. This method was relatively simple, sensitive (less than 1 ?g/kg weight, equivalent to 20 ng/mL), low maintenance, and rapid (less than 60 min).
Cheng, Jinping; Pi, Shuaishuai; Ye, Shufeng; Gao, Haomin; Yao, Lei; Jiang, Zhenyi; Song, Yuling; Xi, Lei
The effect of domestic pan-frying and grilling on Cd, Cr, Cu, Fe, Hg, Ni, Pb and Zn content of popular small Mediterranean finfish and shellfish was studied. The species selected -namely anchovy, bogue, hake, picarel, sardine, sand smelt, stripped mullet, Mediterranean mussel, shrimp and squid- are among the most commonly marketed and consumed in Greece and most of the Mediterranean countries. Both culinary practices examined resulted in increased metals concentrations compared to those of raw samples, the increment being inversely related to fish size and -in most cases- being more extended in pan-frying. The consumption of cooked seafood is expected to provide significant amounts of Fe and Zn followed, in decreasing order, by Cr, Cu and Ni. In addition, the estimation of (a) weekly intakes and (b) target hazard quotients for the toxic elements Cd, Hg and Pb revealed that the cooked fish and shellfish do not pose any health risk for the consumers. PMID:22813872
Kalogeropoulos, Nick; Karavoltsos, Sotirios; Sakellari, Aikaterini; Avramidou, Stella; Dassenakis, Manos; Scoullos, Michael
Vibrio vulnificus is the leading cause of food-related mortality reported in the state of Florida. It is normal microflora in marine environments, where seawater and molluscan shellfish are the primary vectors of V. vulnificus disease. Risk correlates with seasonally high numbers of V. vulnificus bacteria during the summer months. Currently, the infectious dose for humans, as well as whether the disease is caused by single or multiple strains found in molluscan shellfish, is unknown. In this work, we studied pulsed-field gel electrophoresis profiles of V. vulnificus strains isolated from blood and oysters associated with V. vulnificus disease. Results showed that ca. 10(3) V. vulnificus bacteria/gram of oyster and higher concentrations were associated with human infections and that a single V. vulnificus strain, evidenced by pulsed-field gel electrophoresis profiles, was isolated from human tissues.
Jackson, J K; Murphree, R L; Tamplin, M L
Paralytic shellfish poisoning (PSP) toxins are highly toxic natural compounds produced by dinoflagellates commonly present in marine phytoplankton. Shellfish contaminated with these toxins create significant public health threat and economic losses to the shellfish industry. For this reason, several methods of high performance liquid chromatography (HPLC) with fluorescence detection have been developed in order to gain better knowledge of toxins profiles in shellfish and dinoflagellates samples. These methods have been subjected to continuous modifications to improve and shorten the run time of analysis in the routine monitoring control. In this paper, different samples are analyzed by pre- and post- column HPLC methods to compare toxin profiles. All PSP toxins were individually identified and quantified within the post-column oxidation method. However, although the pre-column oxidation method is significantly more sensitive and detects lower toxin levels, it provides a total amount of toxins that co-elute together, as GTX2 and 3, GTX1 and 4 and dcGTX2 and dcGTX3. The results obtained by both HPLC methods showed similar toxin concentration (expressed in mug/mL) in mussel samples, however when dinoflagellates samples were analyzed the toxin profile and concentration were different. In summary, the post-column oxidation method is accurate to determine the amount of each individual PSP toxin and to know the real toxic profile of non-transformed samples. In addition, this method is easy and faster to screen a large number of samples. The pre-column HPLC method is useful when mussel samples are analyzed even though the time required to prepare the samples is longer. PMID:20466012
Rodríguez, P; Alfonso, A; Botana, A M; Vieytes, M R; Botana, L M
Real-time RT-PCR, combining amplification and detection of virus-specific amplicons, is a promising tool for norovirus detection in environmental or food samples such as shellfish. We developed a real-time RT-PCR assay based on one-step detection using single primer sets and probes for norovirus genogroups I and II. Seventy and seven RT-PCR units of genogroup I and II reference norovirus strains, respectively,
F. Loisy; R. L. Atmar; P. Guillon; P. Le Cann; M. Pommepuy; F. S. Le Guyader
Digestive glands containing paralytic shellfish poisoning (PSP) toxins were isolated from toxic scallops. Citrate\\/phosphate buffers with the pH values ranging from 3 to 7 were added to achieve predetermined pH levels. The samples were heated at 90, 100, 110, 120 and 130°C using a computer controlled oil bath, and three tubes at each pH level were transferred into an ice
W. M Indrasena; T. A Gill
In this study, the concentrations of 15 perfluorinated compounds (PFCs) were analyzed in 30 water samples collected in Catalonia (Spain) at three stages of the drinking water treatment process in several water purification plants. In addition, the concentrations of 13 PFCs were determined in samples of fish and shellfish collected from coastal areas of Catalonia. The intake of PFCs through both pathways, drinking water intake and fish and shellfish consumption, was also estimated. In water samples, the highest mean concentrations corresponded to perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) (1.81 and 2.40 ng/L, respectively), whereas perfluorodecanosulfonate (PFDS) and perfluorotetradecanoic acid (PFTDA) were under their respective limits of detection in all analyzed samples. The results show that although the current treatment processes caused slight reductions in PFC concentrations, these processes did not mean significant changes in the amounts of PFCs already contained in the raw water. Among the analyzed PFCs in fish and shellfish, only seven compounds could be detected in at least one composite sample. PFOS showed the highest mean concentration (2.70 ng/g fw), being detected in all species with the exception of mussels. With regard to PFOA (mean, 0.074 ng/g fw), the highest concentrations were detected in prawn and hake (0.098 and 0.091 ng/g fw, respectively). The current exposure to PFCs through consumption of fish and shellfish indicates that it should not be of concern for the consumers. The amounts ingested are well below the recommended tolerable daily intakes, at least for those PFCs for which information is available. PMID:22494245
Domingo, José L; Ericson-Jogsten, Ingrid; Perelló, Gemma; Nadal, Martí; Van Bavel, Bert; Kärrman, Anna
A new analogue of okadaic acid (OA), the toxin mainly responsible for diarrhetic shellfish-poisoning (DSP) phenomena in Europe, has been isolated from toxic phytoplankton (Dinophysis acuta) collected in Irish waters. Fluorimetric LC analyses of the extracts of bulk phytoplankton samples using derivatisation with 9-anthryldiazomethane (ADAM) showed a complex toxin profile, with peaks corresponding to OA and dinophysistoxin-2 (DTX-2) as well
R Draisci; L Giannetti; L Lucentini; C Marchiafava; K. J James; A. G Bishop; B. M Healy; S. S Kelly
The aim of this work was to shed light on the anatomical distribution of diarrhetic shellfish poisoning (DSP) toxins in the mussel Mytilus galloprovincialis and to determine any possible changes undergone during the depuration process. To this end, the distribution of two DSP toxins—okadaic acid and DTX2—and some of their derivatives were studied by means of HPLC\\/MS at different stages
Juan Blanco; Carmen Mariño; Helena Martín; Carmen P. Acosta
Currently, many states use precipitation information to regulate periodic closures of shellfish harvest areas based on a presumptive\\u000a relationship between rainfall and bacteria concentration. We evaluate this relationship in four South Carolina estuaries and\\u000a suggest new predictive models that integrate remote sensing precipitation data with additional environmental and climatic\\u000a data. Model comparisons using Akaike’s information criterion, tenfold cross-validation, and model
Rense Heath Kelsey; Geoffrey I. Scott; Dwayne E. Porter; Thomas C. Siewicki; Donald G. Edwards
The increasing frequency by which the production of paralytic shellfish toxins (PST) by freshwater bloom-forming cyanobacteria is being noticed world-wide raises the possibility of PST bioaccumulation by freshwater mussels. This study evaluates PST accumulation and depuration by the freshwater mussel Anodonta cygnea exposed over a 14-day period to high densities (mean=1.4×109cellsl?1, S.D.=0.29×109cellsl?1) of the toxic cyanobacterium Aphanizomenon issatschenkoi (corresponding to
Paulo Pereira; Elsa Dias; Susana Franca; Elisa Pereira; Manuela Carolino; Vitor Vasconcelos
The increased consumption of fish and shellfish has resulted in more frequent reports of adverse reactions to seafood, emphasizing the need for more specific diagnosis and treatment of this condition and exploring reasons for the persistence of this allergy. This review discusses interesting and new findings in the area of fish and shellfish allergy. New allergens and important potential cross-reacting allergens have been identified within the fish family and between shellfish, arachnids, and insects. The diagnostic approach may require prick to-prick tests using crude extracts of both raw and cooked forms of seafood for screening seafood sensitization before a food challenge or where food challenge is not feasible. Allergen-specific immunotherapy can be important; mutated less allergenic seafood proteins have been developed for this purpose. The persistence of allergy because of seafood proteins' resistance after rigorous treatment like cooking and extreme pH is well documented. Additionally, IgE antibodies from individuals with persistent allergy may be directed against different epitopes than those in patients with transient allergy. For a topic as important as this one, new areas of technological developments will likely have a significant impact, to provide more accurate methods of diagnosing useful information to patients about the likely course of their seafood allergy over the course of their childhood and beyond. PMID:22554093
Tsabouri, Sophia; Triga, Maria; Makris, Michael; Kalogeromitros, Dimitris; Church, Martin K; Priftis, Kostas N
The objectives of this study were to document the spatial and temporal distributions and compositions of bacteria in the sediments and overlying waters of an important urban shellfishing area in the lower Chesapeake Bay region, the Lynnhaven Estuary. Marked fluctuations were observed in the date of many of the physicochemical parameters and the indicator bacteria. The higher-salinity water and coarser sediment of the inlet site showed lower overall bacterial densities than did the headwater sites, where freshwater runoff and decreased tidal action were characteristic. Densities of benthic indicator bacteria, when expressed on a volumetric basis, were significantly greater than counts in the overlying waters. These counts were indicative of a fecally polluted system and were well above the safe maximum limits for shellfish-growing waters. Significantly fewer total and fecal bacteria were observed in both the water and the sediment during the warm months of May, July, and August. The primary sources of the Lynnhaven's bacterial pollution appeared to be typical of urban and agricultural runoff, although failure of septic tank systems was suspected as a problem in the Lynnhaven's western branch. These results illustrated that sediments in shellfishing areas could serve as a reservoir for high densities of indicator bacteria and that, potentially, pathogens could pose a health hazard.
Erkenbrecher, C W
A new ELISA kit to quantitate the level of paralytic shellfish poisoning (PSP) toxins in crude shellfish extracts was developed. A conjugate for preparing antigen and a novel antibody used in the ELISA was prepared based on the unique reactions between C11-O-sulfate toxins such as gonyautoxins 2 and 3 (GTX2,3) and various thiol compounds, followed by coupling to keyhole limpet hemocyanin. The compounds necessary for competitive ELISA, labeled toxin and an artificial standard toxin to replace saxitoxin in the analysis, were also produced by the same techniques. The resulting ELISA recognized all the toxin components tested; however, carbamoyl-N-sulfate derivatives such as B and C toxins and N1-OH toxins such as neoSTX and GTX1,4 showed low affinity to the antibody. The difference in the reactivity of the antibody observed among the toxin components prevents accurate quantification of the toxin amounts in shellfish extracts. To address this problem, the former toxin components were transformed to corresponding carbamate toxins by mild HCl treatment according to a conventional method. The reduction of N1-OH of the latter toxins to N1-H was performed by our original method using hemin as a catalyst. We report here the new ELISA kit coupled with the pretreatment process to transform the toxin components favorable for the quantitative analysis of PSP toxins. PMID:24830145
Sato, Shigeru; Takata, Yoshinobu; Kondo, Sunaho; Kotoda, Akiko; Hongo, Naoto; Kodama, Masaaki
At present, the analytical method for paralytic shellfish poisoning (PSP) toxins in shellfish is the mouse bioassay (MBA), which is an official method of the Association of Analytical Communities (AOAC ). However, the low sensitivity and concerns over the number of live animals required for testing have been cited as the major reason for seeking its replacement. In this report, we employed an open-sandwich immunoassay (OS-IA) to detect gonyautoxin (GTX2/3), a kind of PSP toxins. OS-IA, which utilizes the antigen-induced enhancement of antibody VH/VL interaction, can measure a small molecule antigen in a noncompetitive format. Hence it has a wider working range and shorter measurement time. We isolated anti-GTX2/3 antibody gene from a hybridoma GT-13A by screening a Fab-displaying phage library. Then the vectors for OS-IA were constructed, and examined for antigen concentration-dependency of the VH/VL interaction by OS-ELISA. As a result, in each case, signal intensity increases notably in a wide concentration range (0.1 to >1000 ng mL(-1)) of free GTX2/3, which was enough to cover its regulation value (80 ?g 100 g(-1)) in many countries. So OS-IA will be widely applicable to detect PSP toxins in shellfish meats and in drinking water. PMID:23953213
Hara, Yuko; Dong, Jinhua; Ueda, Hiroshi
Viruses are the leading cause of foodborne illness associated with the consumption of raw or slightly-cooked contaminated shellfish. This study evaluated the E.Z.N.A. Mollusc RNA extraction and purification kit for the detection of HAV in shellfish. The E.Z.N.A. method, based on the cationic detergent, cetyltrimethyl ammonium bromide, in conjunction with a selective RNA binding silica matrix, efficiently isolated viral RNA with a detection limit of 1TCID(50)/ml by hemi-nested PCR. This method proved to be faster and less expensive than PEG precipitation-based procedures. It is also technically undemanding, requiring no extensive processing steps or excess manipulation, minimizing RNA degradation and ensuring the absence of PCR inhibitors. The E.Z.N.A. method applied to HAV screening of shellfish samples from the Apulian region, revealed a high level of contamination. These results confirm that conventional faecal indicators are unreliable for demonstrating the presence or absence of viruses. PMID:20638741
Terio, Valentina; Di Pinto, Angela; Di Pinto, Pietro; Martella, Vito; Tantillo, Giuseppina
Shellfish have the capacity to accumulate chemical contaminants found in their biotope and therefore present a potential risk for consumers. This study was conducted to assess the chemical risks associated with consumption of shellfish harvested on the north shore of the St. Lawrence River's lower estuary. A survey was carried out on 162 recreational harvesters, and shellfish were sampled for chemical contaminant analysis. We quantified 10 metals, 22 polycyclic aromatic hydrocarbons (PAHs), 14 polychlorinated biphenyls (PCBs), and 10 chlorinated pesticides. We subsequently evaluated cancer and noncancer risks for four consumption scenarios based on our survey results and published results. Soft-shell clams (Mya arenaria) were by far the most consumed shellfish species. Of the 56 selected contaminants, 36 were detected in the 23 homogenates of soft-shell clam meat. None of the contaminants found in the soft-shell clams were associated with intakes that exceed the main exposure limit recommendations proposed to prevent noncancer effects. However, several limits must be considered before drawing conclusions about the relative safety of shellfish consumption regarding this end point. Furthermore, inorganic arsenic and PCBs were present in sufficient concentrations to lead to cancer risks exceeding the level often considered acceptable for environmental exposure (1 x 10 (-4) to 1 x 10(-6)) in each of the four scenarios, even for the lowest observed scenario of 15 meals of soft-shell clams per year.
Gagnon, Fabien; Tremblay, Thierry; Rouette, Justine; Cartier, Jacques-Francois
Shellfish have the capacity to accumulate chemical contaminants found in their biotope and therefore present a potential risk for consumers. This study was conducted to assess the chemical risks associated with consumption of shellfish harvested on the north shore of the St. Lawrence River's lower estuary. A survey was carried out on 162 recreational harvesters, and shellfish were sampled for chemical contaminant analysis. We quantified 10 metals, 22 polycyclic aromatic hydrocarbons (PAHs), 14 polychlorinated biphenyls (PCBs), and 10 chlorinated pesticides. We subsequently evaluated cancer and noncancer risks for four consumption scenarios based on our survey results and published results. Soft-shell clams (Mya arenaria) were by far the most consumed shellfish species. Of the 56 selected contaminants, 36 were detected in the 23 homogenates of soft-shell clam meat. None of the contaminants found in the soft-shell clams were associated with intakes that exceed the main exposure limit recommendations proposed to prevent noncancer effects. However, several limits must be considered before drawing conclusions about the relative safety of shellfish consumption regarding this end point. Furthermore, inorganic arsenic and PCBs were present in sufficient concentrations to lead to cancer risks exceeding the level often considered acceptable for environmental exposure (1 x 10 (-4) to 1 x 10(-6)) in each of the four scenarios, even for the lowest observed scenario of 15 meals of soft-shell clams per year. PMID:15175177
Gagnon, Fabien; Tremblay, Thierry; Rouette, Justine; Cartier, Jacques-François
Recent efforts to restore oyster reefs have resulted in the creation of many reefs with the explicit objective of benefiting local shellfish and finfish fisheries. We evaluated the community responses of the fish (transient and resident), crab, and shellfish species that colonized or utilized a series of restored high- and low-relief oyster reefs at three different locations within Mobile Bay,
Kevan C. Gregalis; Matthew W. Johnson; Sean P. Powers
A comparative study was conducted to determine the feasibility of enzyme-linked immunosorbent assays (ELISAs) for the detection of amnesic shellfish poisoning (ASP) and paralytic shellfish poisoning (PSP) toxins in nine naturally contaminated species in fresh, frozen, boiled and canned fish and shellfish. PSP and ASP were analyzed in 138 shellfish samples (mussels, clams, barnacles, razor shells, scallops and cockles) and anchovies by mouse bioassay (MBA) and high performance liquid chromatography with ultraviolet detection (HPLC-UV), respectively. Results were compared with toxin concentrations obtained using two commercial competitive ELISAs, saxitoxin and ASP kits. Immunoassays were able to quantify toxins in different matrices showing excellent Pearson's correlation coefficients (r = 0.974 for saxitoxin ELISA and r = 0.973 for ASP ELISA) and to detect PSP and ASP with a lower limit of detection (LOD), namely, 50 microg saxitoxin equivalent/kg shellfish meat for PSP and 60 microg/kg domoic acid in shellfish flesh for ASP, than the reference methods (350 microg saxitoxin equivalent/kg shellfish meat and 1.6 mg/kg domoic acid in shellfish flesh, respectively). These results suggest that the ELISA method could be used as screening systems in a variety of species without matrix interference. PMID:20088511
Garet, Elina; González-Fernández, Africa; Lago, Jorge; Vieites, Juan M; Cabado, Ana G
Aquaculture is viewed as a potential mechanism to meet the growing demand for seafood around the world. The future of bivalve shellfish aquaculture in the U.S. hinges on sustainable practices on the part of industry and a more consistent regulatory regime. Bivalve shellfish aquaculture is a recent practice relative to its history in other countries, beginning in the late 1800s
Brett R. Dumbauld; Jennifer L. Ruesink; Steven S. Rumrill
Uptake and depuration kinetics and BCFs of several pesticides in three species of shellfish ( Corbicula leana, Corbicula japonica, and Cipangopludina chinensis): Comparison between field and laboratory experiment
Uptake and depuration rate constants, and bioconcentration factors (BCFs) of pesticides in shellfish (Corbicula leana, Corbicula japonica and Cipangopludina chinensis) were estimated from both field and laboratory experiments. The simulation of pesticide concentrations in shellfish in the field using the rate constants obtained from the laboratory experiments and the actual concentrations of pesticides in the river water, roughly predicted the
Seiichi Uno; Hiroaki Shiraishi; Shigehisa Hatakeyama; Akira Otsuki
Since organochlorine compounds are known to be accumulated in benthic animals and CNP is very persistent in aquatic animals, shellfish might be useful as an indicator of environmental contamination by CNP. In order to understand the correlations between the concentrations in shellfish, water and sediments, it is necessary to investigate their temporal changes and the residue half-life time of CNP in the environment. For this purpose, CNP-free shellfish (Corbicula japonica) were transferred to fixed point in the lower reaches of the Ishikari River, and the CNP concentrations in shellfish, water and sediments, before and after CNP application, were examined biweekly from May to August and monthly from September to December 1984. The present paper will show that shellfish can be a biological indicator for CNP contamination in the river.
Marine phycotoxins derived from harmful algal blooms are known to be associated with mass mortalities in the higher trophic levels of marine food webs. Bivalve mollusks and planktivorous fish are the most studied vectors of marine phycotoxins. However, field surveys recently showed that cephalopod mollusks also constitute potential vectors of toxins. Thus, here we determine, for the first time, the time course of accumulation and depuration of paralytic shellfish toxins (PSTs) in the common octopus (Octopus vulgaris). Concomitantly, the underlying kinetics of toxin transfer between tissue compartments was also calculated. Naturally contaminated clams were used to orally expose the octopus to PSTs during 6 days. Afterwards, octopus specimens were fed with non-contaminated shellfish during 10 days of depuration period. Toxins reached the highest concentrations in the digestive gland surpassing the levels in the kidney by three orders of magnitude. PSTs were not detected in any other tissue analyzed. Net accumulation efficiencies of 42% for GTX5, 36% for dcSTX and 23% for C1+2 were calculated for the digestive gland. These compounds were the most abundant toxins in both digestive gland and the contaminated shellfish diet. The small differences in relative abundance of each toxin observed between the prey and the cephalopod predator indicates low conversion rates of these toxins. The depuration period was better described using an exponential decay model comprising a single compartment - the entire viscera. It is worth noting that since octopuses' excretion and depuration rates are low, the digestive gland is able to accumulate very high toxin concentrations for long periods of time. Therefore, the present study clearly shows that O. vulgaris is a high-potential vector of PSTs during and even after the occurrence of these toxic algal blooms. PMID:24316438
Lopes, Vanessa M; Baptista, Miguel; Repolho, Tiago; Rosa, Rui; Costa, Pedro Reis
Background Microsatellite DNA is one of many powerful genetic markers used for the construction of genetic linkage maps and the study of population genetics. The biological databases in public domain hold vast numbers of microsatellite sequences for many organisms including fishes. The microsatellite data available in these data sources were extracted and managed into a database that facilitates sequences analysis and browsing relevant information. The system also helps to design primer sequences for flanking regions of repeat loci for PCR identification of polymorphism within populations. Description FishMicrosat is a database of microsatellite sequences of fishes and shellfishes that includes important aquaculture species such as Lates calcarifer, Ctenopharyngodon idella, Hypophthalmichthys molitrix, Penaeus monodon, Labeo rohita, Oreochromis niloticus, Fenneropenaeus indicus and Macrobrachium rosenbergii. The database contains 4398 microsatellite sequences of 41 species belonging to 15 families from the Indian subcontinent. GenBank of NCBI was used as a prime data source for developing the database. The database presents information about simple and compound microsatellites, their clusters and locus orientation within sequences. The database has been integrated with different tools in a web interface such as primer designing, locus finding, mapping repeats, detecting similarities among sequences across species, and searching using motifs and keywords. In addition, the database has the ability to browse information on the top 10 families and the top 10 species, through record overview. Conclusions FishMicrosat database is a useful resource for fish and shellfish microsatellite analyses and locus identification across species, which has important applications in population genetics, evolutionary studies and genetic relatedness among species. The database can be expanded further to include the microsatellite data of fishes and shellfishes from other regions and available information on genome sequencing project of species of aquaculture importance.
During the last few years the occurrence of a high percentage of esters of diarrhetic shellfish poisoning (DSP) toxins has been observed in shellfish from the Portuguese coast. Most of the commercial bivalves contain DSP toxins in ester forms, either acyl derivatives of okadaic acid (OA) or of dinophysistoxin-2 (DTX-2). The stability of these toxins in shellfish tissues and in raw methanol extracts was investigated in two different naturally contaminated species, mussel and carpet shell, over a 4-week period. The results for both species revealed that DSP toxins were more stable in tissue than in raw methanol extracts. Losses of DSP toxins were seen in the first 2 weeks and were more than 30%, but after that a period of stabilization was observed. The decrease was due probably from losses of esters of OA and DTX-2, the free toxins were stable over the period studied. The extraction most commonly used for chemical and biochemical assays relied on methanolic extraction with aqueous 80% methanol. In this work we have tested the extraction solvent on the extractability of DSP toxins from several naturally contaminated species. A single dispersive extraction with methanol, with solvent ratios of 70%, 80%, 90% and 100%, were tested. After alkaline hydrolysis of esterified toxins and clean-up with hexane and dichloromethane, the samples were analysed by liquid chromatography-mass spectrometry (LC-MS). The recovery of DSP toxins increased with increasing percentages of methanol up to 90%. A decrease in recovery with 100% methanol was observed probably due to problems during the liquid-liquid partitioning. PMID:19680894
Rodrigues, S M; Vale, P; Botelho, M J; Gomes, S S
Vibrio parahaemolyticus (Vp) is found naturally in coastal saltwater. In the United States, Vp causes an estimated 35,000 domestically acquired foodborne infections annually, of which most are attributable to consumption of raw or undercooked shellfish. Illness typically consists of mild to moderate gastroenteritis, although severe infection can occur. Demographic, clinical, and exposure information (including traceback information on implicated seafood) for all laboratory-confirmed illnesses are reported by state health departments to CDC through the Cholera and Other Vibrio Surveillance system. Vp isolates are distinguished by serotyping (>90 serotypes have been described) and by pulsed-field gel electrophoresis (PFGE). PMID:24739344
Newton, Anna E; Garrett, Nancy; Stroika, Steven G; Halpin, Jessica L; Turnsek, Maryann; Mody, Rajal K
Background Shellfish can be a component of a healthy diet due to a low fat and high protein content, but the cholesterol content of some species is often cited as a reason to limit their consumption. Data on levels of non-cholesterol sterols in commonly consumed species are lacking. Objective Shellfish were sampled and analyzed to update sterol data in the United States Department of Agriculture (USDA) National Nutrient Database for Standard Reference. Design Using a nationwide sampling plan, raw shrimp and sea scallops, canned clams, and steamed oysters, blue crab, and lobster were sampled from 12 statistically selected supermarkets across the United States in 2007–08. For each species, four composites were analyzed, each comprised of samples from three locations; shrimp and scallops from six single locations were also analyzed separately. Using validated analytical methodology, 14 sterols were determined in total lipid extracts after saponification and derivatization to trimethylsilyethers, using gas chromatography for quantitation and mass spectrometry for confirmation of components. Results Crab, lobster, and shrimp contained significant cholesterol (96.2–27 mg/100 g); scallops and clams had the lowest concentrations (23.4–30.1 mg/100 g). Variability in cholesterol among single-location samples of shrimp was low. The major sterols in the mollusks were brassicasterol (12.6–45.6 mg/100 g) and 24-methylenecholesterol (16.7–41.9 mg/100 g), with the highest concentrations in oysters. Total non-cholesterol sterols were 46.5–75.6 mg/100 g in five single-location scallops samples, but 107 mg/100 g in the sixth, with cholesterol also higher in that sample. Other prominent non-cholesterol sterols in mollusks were 22-dehydrocholesterol, isofucosterol, clionasterol, campesterol, and 24-norcholesta-5,22-diene-3?-ol (4–21 mg/100 g). Conclusions The presence of a wide range of sterols, including isomeric forms, in shellfish makes the analysis and quantitation of sterols in marine species more complex than in animal and plant tissues. The detailed sterol composition reported herein provides data that may be useful in research on the impact of shellfish consumption on dietary risk factors.
Phillips, Katherine M.; Ruggio, David M.; Exler, Jacob; Patterson, Kristine Y.
Vertical distributions of dissolved species across the sediment-water interface (SWI), including major cations (sodium, potassium, magnesium, calcium), minor cations (lithium, strontium, barium), redox sensitive species (dissolved manganese, iron, sulfate, sulfide, ammonium) and other chemical parameters (pH, alkalinity, soluble reactive phosphorous, dissolved silica) were studied in a Mediterranean lagoon used for intensive shellfish farming. In order to quantify the impact of this activity on diagenetic processes and the influence of seasonal changes, two stations contrasted with respect to organic carbon fluxes were sampled in Thau lagoon from March 2001 to August 2002 during four field campaigns in winter, spring, summer and fall. Well-defined layers enriched with redox sensitive species were observed following the conventional sequence of early diagenetic reactions. However, differences were observed between both stations in depths and thickness layers. Concentration gradients extended down to more than 92 cm depth at the central position of the lagoon (station C4 - 8 m depth) and down to 40 cm depth inside shellfish farming zones (station C5 - 9 m depth). Station C4 showed an unusual diagenetic signature: sharp dissolved oxygen, iron, nitrate and manganese gradients existed at the SWI but gradients of dissolved sulfide and alkalinity as well as other parameters (dissolved silica, Ba, etc.) were recorded only from 25 to 30 cm depth downward. Seasonal changes were observed in pore water composition as deep as 30-50 cm in station C4 (only 15 cm in station C5). The center of the lagoon is not directly subjected to biodeposits deriving from shellfish activity. Isotopic and bioturbation data allowed to rule out a reworking of the sediment deeper than a few centimeters. In addition to organic content of the sediment, physical parameters were likely to induce the 10-20 cm gap between dissolved iron and sulfide profile as well as the higher vertical extent of diagenetic sequence observed at station C4. Conversely to station C5, station C4 underwent stronger currents and wave effect probably generating advective transport of water through the sediment, but no permeability data were available to confirm this hypothesis. During summer, climatic conditions generated vertical stratification of the water column and transient suboxic conditions at the bottom. Such conditions drove the upward shift of redox fronts, compacting the diagenetic sequence. These effects were reinforced at station C5 by shellfish and its farm structures (mainly attenuation of current and increased heat absorption).
Metzger, E.; Simonucci, C.; Viollier, E.; Sarazin, G.; Prévot, F.; Jézéquel, D.
Paralytic shellfish poisoning (PSP) is the foodborne intoxication associated with the consumption of seafood contaminated with naturally occurring neurotoxins known as paralytic shellfish toxins. To protect public health from this potentially fatal syndrome, harvesting closures are implemented when toxins exceed the regulatory action level. Traditional monitoring programs established by state shellfish authorities allow for timely closures in state waters with minimal negative impacts on industry. However, such monitoring programs are not feasible in federal offshore waters given their distance from shore and the range of their spatial coverage. Thus innovative management strategies were investigated for these offshore resources. Georges Bank, an offshore resource with an estimated market value of more than 3 billion in Atlantic surfclams and ocean quahogs, has been closed to harvesting following a temporary ban in 1989 and a subsequent indefinite closure in 1990 due to the risk of PSP. As a means of managing this risk and allowing harvest of safe shellfish from this important resource, the Onboard Screening Dockside Testing Protocol (referred to as the Protocol) was developed by the US Food and Drug Administration (FDA), National Marine Fisheries Service (NMFS), state shellfish control authorities, and industry. The Protocol, which sets forth control measures to ensure product safety and public health protection, was endorsed by the Interstate Shellfish Sanitation Conference (ISSC) for pilot testing. Briefly, the pilot study Protocol required that (1) the fishing vessel receive a permit from NMFS to harvest in closed waters, (2) a mini?m of five shellfish samples per intended harvest lot be tested for PSP toxins onboard, and (3) harvesting only occur when the samples tested from the intended fishing area are negative using the Jellett Rapid Tests or Abraxis Shipboard ELISA kits. Finally, product landed under the Protocol was confirmed to be safe for consumption using the mouse bioassay (MBA) prior to its introduction into commerce. This paper presents data from the pilot study, with primary focus on the advantages and challenges of the field kits employed onboard compared to the dockside MBA, which has served as the longstanding regulatory method for PSP toxins. In 2010 alone, the successful pilot study resulted in the safe harvest of over 2.7 million worth of surfclams in an area that has otherwise been unavailable for decades. Due to the success of this pilot study, the Protocol was adopted into the National Shellfish Sanitation Program Model Ordinance as an approved marine biotoxin control strategy for use in federal waters at the 2011 ISSC Biennial Meeting. In January 2013 a portion of Georges Bank was reopened for the harvest of Atlantic surfclams and ocean quahogs to fishermen following the Protocol.
DeGrasse, Stacey; Conrad, Stephen; DiStefano, Paul; Vanegas, Camilo; Wallace, David; Jensen, Pete; Hickey, J. Michael; Cenci, Florence; Pitt, Jaclyn; Deardorff, Dave; Rubio, Fernando; Easy, Dorothy; Donovan, Mary Anne; Laycock, Maurice; Rouse, Debbie; Mullen, John
During the week of 14-20 January 2000, 120 people visited the Emergency Departments of hospitals in Thessaloniki, northern Greece, complaining of acute gastrointestinal illness after eating mussels (Mytilus galloprovincialis). The symptoms indicated diarrhoeic shellfish poisoning, and the toxicity of mussels harvested from Thermaikos Gulf in Thessaloniki during the outbreak was investigated using mouse bioassays. The bioassays revealed toxicity to mice by the mussel samples; while high numbers of toxic algae Dinophysis acuminata were identified in water samples from Thermaikos Gulf. The harvesting of mussels was immediately suspended and a monitoring programme for algal blooms was established from then onwards. During a follow-up of the mussels' toxicity from January 2000 to January 2005, two more mussel samples were found positive for diarrheic shellfish poisoning: one harvested in March 2001 from the area of the outbreak (Thermaikos Gulf) and the other harvested in January 2001 from Amvrakikos Gulf in north-western Greece. However, no sporadic cases or outbreaks were reported during this period. PMID:17364933
Economou, V; Papadopoulou, C; Brett, M; Kansouzidou, A; Charalabopoulos, K; Filioussis, G; Seferiadis, K
Human enteric viruses are largely excreted in faeces. These resistance of these viruses in the environment makes their faecal-oral transmission easier. Filter feeder organisms such as the mussels are bio-accumulators of viruses contaminating their aquatic environment. Thus, undercooked shellfish consumption involves sanitary risks. Thirty samples of mussels (Mytilus sp.), were tested, half were from an aquaculture origin, the others were from an area more exposed to faecal pollution. Fifteen sewage samples from this last area were also examined. Viruses were extracted from the digestive tissue by direct elution method in a glycine/NaCl pH 9.5 buffer followed by PEG 8000 precipitation. The PEG pellets were used for DNA extraction by proteinase K and phenol/chloroform. The molecular characterization, by PCR using specific adenovirus primers revealed that shellfish growing on Mohammedia (a town in the Casablanca outskirts) littoral are contaminated whereas those chosen from aquaculture and bought in the central market were not contaminated. PMID:15847929
Karamoko, Y; Ibenyassine, K; Aitmhand, R; Idaomar, M; Ennaji, M M
Paralytic shellfish toxin concentrations were measured and individual toxin profiles were monitored in axenic batch cultures of Protogonyaulax tamarensis and Protogonyaulax catenella. High pressure liquid chromatographic methods were used that allowed the separation of all 12 known paralytic shellfish poisons, including toxins C1, C2, and C3, from a single sample. In isolates of both Protogonyaulax species, total toxin levels were relatively low after inoculation, increased rapidly in early to mid-exponential growth to a value 100 to 300% of that at the initial time point, then decreased by 86 to 95% as the culture aged. Although the concentrations of individual toxins per cell followed the same general pattern as that seen for total moles of toxin per cell, variability in toxin profile with culture age was observed. In P. tamarensis, the mole percent of neosaxitoxin increased substantially from 8 to 44% as total toxin levels per cell decreased. A concomitant decrease in the mole percent of saxitoxin with culture age was noted. Although not as precipitous, changes in the mole percent of specific toxins from P. catenella were also observed. The mole percent of gonyautoxins I and IV increased, while that of gonyautoxins II and III decreased. These data suggest that the toxin profile in isolates of Protogonyaulax can change, sometimes significantly, with changing environmental variables.
Boczar, Barbara A.; Beitler, Mark K.; Liston, John; Sullivan, John J.; Cattolico, Rose Ann
Tributyltin (TBT) and triphenyltin (TPT) compounds have been widely used as antifouling paint for ship bottoms and fishery firm nets, and they are known to be aquatic environmental contaminants. A survey of the pollution with organotin compounds in fish and shellfish has been carried out for samples (180 samples from 43 kinds) which were collected in the Hokkaido prefecture during the period from February, 1989 to September, 1992. On the other hand, daily intakes of organotin compounds have been investigated using the Market Basket Method. Relatively high concentrations of TBT and TPT were detected from flat fish and shellfish such as Hirame (bastard halibut), Kurogarei (black plaice), Asari (Japanese littleneck), Hokkigai (Japanese surf clam) and Kaki (oyster). The maximum value of TBT in samples was 0.38 microgram/g (for both Asari and Kaki), and TPT was 0.47 microgram/g (Kurogarei). Estimated daily intakes of DBT, TBT and TPT were 0.45 microgram, 2.40 micrograms and 4.11 micrograms, respectively. The values presented in this study were lower in comparison with the acceptable daily intake. It was revealed that the values of the organotin compounds in this study were not so high as to have any efect on human health at the present. PMID:8157252
This paper describes an outbreak of paralytic shellfish poisoning (PSP), affecting human populations on the Pacific Coast of Costa Rica in October 1989. Numbness in arms, face and legs occurred 30 to 45 minutes after ingestion of the large clam Spondylus calcifer. Paralysis of legs and respiratory symptoms followed, often persisting for one week. Large amounts of the dinoflagellate Pyrodinium bahamense were found in the intestine of the mollusk. A toxin was detected in crude or filtered and heated macerates of intestine, muscle, mantle and hepatopancreas of S. calcifer, and to a lesser extent Tagelus sp., by injection of its crude or diluted extracts in white mice. The effects in mice consisted in paralysis and asphyxia generally leading to death in less than 5 minutes, compatible with saxitoxin. Mice were killed by the toxin in macerates diluted 1:100 to 1:1000. No toxin was detected in Anadara tuberculosa (Bivalvia) or in peneids. Prevention rests on intersectoral actions between state and private sectors in charge of fishing, distribution and marketing of shellfish, as well as on education of the population at large. PMID:2093952
Mata, L; Abarca, G; Marranghello, L; Víquez, R
To prevent the consumption of bivalves contaminated with paralytic shellfish poisoning (PSP), toxin levels in seafood products are estimated by using the official mouse bioassay. Because of the limitations of this bioassay other methods of monitoring toxins are clearly needed. We have developed a test to screen for PSP toxins based on its functional activity; the toxins bind to the voltage-gated Na+ channels and block their activity. The method is a fluorimetric assay that allows quantitation of the toxins by detecting changes in the membrane potential of human excitable cells. This assay gives an estimate of toxicity, since each toxin present in the sample binds to sodium channels with an affinity which is proportional to its intrinsic toxic potency. The detection limits for paralytic shellfish toxins were found to be 1 ng saxitoxin equivalents/ml compared to the regulatory limit threshold of 400 ng/ml (equivalent to 80 microg/100 g) used in most countries. Our results indicate that this fluorescent assay is a specific, very sensitive, rapid, and reliable method of monitoring PSP toxin levels in samples from seafood products and toxic algae. PMID:11161318
Louzao, M C; Vieytes, M R; Baptista de Sousa, J M; Leira, F; Botana, L M
Temperature regulation is a vital component of any aquaculture system. Existing facilities can be retrofitted with extra insulation, waste heat recovery systems and in some cases, active solar water heating. Those aquaculture ventures that seek to raise organisms to market size under controlled conditions are currently hindered by high operating costs, including fuel. These outfits can also benefit from conservation and alternative energy technologies. In addition, the industry may be more willing to cultivate species normally restricted by climatic conditions if a less expensive source of heating were available. This report focuses on three of the bivalve shellfish aquaculture enterprises of Long Island, New York. In the 1978 to 1979 growing season, Long Island shellfish growers collectively burned over 50,000 gallons of heating oil to warm their hatchery waters and buildings. Since then, heating oil prices have doubled. Currently, some growers are limited by these fuel costs from beginning production earlier in the season. In this report, several heat conservation measures are discussed, and the feasibility of active solar hot water is examined.
Paralytic shellfish toxins (PSTs) are a risk to humans upon consumption of contaminated seafood. The PST family is comprised of more than twenty congeners, with each form having a different potency. In order to adequately protect consumers yet reduce unnecessary closures of non-contaminated harvesting areas, a rapid method that allows for analysis of sample toxicity is needed. While a number of PST immunoassays exist, the outstanding challenge is linking quantitative response to sample toxicity, as no single antibody reacts to the PST congeners in a manner that correlates with potency. A novel approach, then, is to combine multiple antibodies of varying reactivity to create a screening assay. This research details our investigation of three currently available antibodies for their reactivity profiles determined using a surface plasmon resonance biosensor assay. While our study shows challenges with detection of the R1-hydroxylated PSTs, results indicate that using multiple antibodies may provide more confidence in determining overall toxicity and the toxin profile. A multiplexed approach would not only improve biosensor assays but could also be applied to lateral flow immuno-chromatographic platforms, and such a theoretical device incorporating the three antibodies is presented. These improved assays could reduce the number of animal bioassays and confirmatory analyses (e.g., LC/MS), thereby improving food safety and economic use of shellfish resources. PMID:22967609
Yakes, Betsy Jean; Prezioso, Samantha M; DeGrasse, Stacey L
The objective of the Hanford Environmental Dose Reconstruction Project (HEDR) is to estimate the potential radiation doses received by people living within the sphere of influence of the Hanford Site. A potential critical pathway for human radiation exposure is through the consumption of waterfowl that frequent onsite waste-water ponds or through eating of fish, shellfish, and waterfowl that reside in/on the Columbia River and its tributaries downstream of the reactors. This document summarizes information on fish, shellfish, and waterfowl radiation contamination for samples collected by Hanford monitoring personnel and offsite agencies for the period 1945 to 1972. Specific information includes the types of organisms sampled, the kinds of tissues and organs analyzed, the sampling locations, and the radionuclides reported. Some tissue concentrations are also included. We anticipate that these yearly summaries will be helpful to individuals and organizations interested in evaluating aquatic pathway information for locations impacted by Hanford operations and will be useful for planning the direction of future HEDR studies.
Hanf, R.W.; Dirkes, R.L.; Duncan, J.P.
Mussels filter large volumes of water and can concentrate pathogenic organisms, which may act as potential vehicles of transmission to the consumer. A survey study was carried out to investigate the presence of Cryptosporidium protozoan parasites in green mussels (Perna viridis), the smussles pecies most destined for consumption in Thailand. In total, 56 samples were examined from Bangkok (n = 24) and Samut Prakan (n = 32) a wholesale shell-fish markets located at the mouth of the Chao Phraya River. The market for green mussels was closed to the mussel culture placed along the coastal line and this localization may have significant economical impact if the mussels' cultures are found contaminated. Cryptosporidium spp. oocysts were detected by the immunofluorescence antibody method (IFA) in 12.5% of the samples examined. The detection of Cryptosporidium oocysts in green mussels' population of Samut Prakan was higher (15.6%) than in Bangkok market (8.3%). These differences in positive samples from the two locations may be caused by physical, ecological and anthropogenic conditions. This could relay to different contamination levels of marine water by Cryptosporidium oocysts and consequently to contamination of harvested shellfish populations. The results demonstrate that the Cryptosporidium spp. oocysts were found indigenous in mussels from the coastal line of Thailand, indicating that mussels may act as a reservoir of Cryptosporidium foodborne infections for humans. PMID:19839271
Srisuphanunt, M; Wiwanitkit, Viroj; Saksirisampant, W; Karanis, P
Hepatitis E virus (HEV), an enteric pathogen of both humans and animals, is excreted by infected individuals and is therefore present in wastewaters and coastal waters. As bivalve molluscan shellfish are known to concentrate viral particles during the process of filter feeding, they may accumulate this virus. The bioaccumulation efficiencies of oysters (Crassostrea gigas), flat oysters (Ostrea edulis), mussels (Mytilus edulis), and clams (Ruditapes philippinarum) were compared at different time points during the year. Tissue distribution analysis showed that most of the viruses were concentrated in the digestive tissues of the four species. Mussels and clams were found to be more sensitive to sporadic contamination events, as demonstrated by rapid bioaccumulation in less than 1 h compared to species of oysters. For oysters, concentrations increased during the 24-h bioaccumulation period. Additionally, to evaluate environmental occurrence of HEV in shellfish, an environmental investigation was undertaken at sites potentially impacted by pigs, wild boars, and human waste. Of the 286 samples collected, none were contaminated with hepatitis E virus, despite evidence that this virus is circulating in some French areas. It is possible that the number of hepatitis E viral particles discharged into the environment is too low to detect or that the virus may have a very short period of persistence in pig manure and human waste. PMID:24795382
Grodzki, Marco; Schaeffer, Julien; Piquet, Jean-Côme; Le Saux, Jean-Claude; Chevé, Julien; Ollivier, Joanna; Le Pendu, Jacques; Le Guyader, Françoise S
The uptake of paralytic shellfish poisoning (PSP) toxins and spirolides by the paddle crab (Ovalipes catharus) was investigated in two laboratory feeding trials using Greenshell™ mussels (Perna canaliculus), which had been fed toxic strains of either Alexandrium catenella or A. ostenfeldii, as a vector. Toxin uptake by crabs occurred in both feeding trials and was limited to the visceral tissue;
Rozalind Jester; Lesley Rhodes; Veronica Beuzenberg
The neurotoxin ?-N-methylamino-l-alanine (BMAA) and its putative role in multiple neurodegenerative diseases have been intensely studied since 2005 when the toxin was discovered to be produced by worldwide-distributed cyanobacterial species inhabiting terrestrial, marine, brackish, and freshwater ecosystems. Recently, BMAA production was also associated with one eukaryotic group, namely, diatoms, raising questions about its production by other phytoplanktonic groups. To test for BMAA bioavailability in ecosystems where abundant phytoplanktonic blooms regularly occur, samples of filter-feeding shellfish were collected in two Portuguese transitional water bodies. BMAA content in cockles (Cerastoderma edule) collected weekly between September and November 2009 from Ria de Aveiro and at least once a month from May to November from Ria Formosa, fluctuated from 0.079±0.055 to 0.354±0.066?g/g DW and from below the limit of detection to 0.434±0.110?g/g DW, respectively. Simultaneously to BMAA occurrence in cockles, paralytic shellfish toxins were detected in shellfish as a result of Gymnodinium catenatum blooms indicating a possible link between this marine dinoflagellate and BMAA production. Moreover, considerable high BMAA levels, 0.457±0.186?g/g DW, were then determined in a laboratory grown culture of G. catenatum. This work reveals for the first time the presence of BMAA in shellfish from Atlantic transitional water bodies and consubstantiate evidences of G. catenatum as one of the main sources of BMAA in these ecosystems. PMID:24747603
Lage, Sandra; Costa, Pedro Reis; Moita, Teresa; Eriksson, Johan; Rasmussen, Ulla; Rydberg, Sara Jonasson
Procedures have been developed for the extraction and high-performance liquid chromatography (HPLC) analysis of paralytic shellfish poisoning (PSP) toxins from Protogonyaulax spp. grown in batch culture. Using these procedures, the toxin content of two isolates of P. tamarensis (NEPCC 183 and 255) and one isolate of P. catenella (NEPCC 355) were examined. Total toxin and individual toxin concentrations were measured
G. L. Boyer; J. J. Sullivan; R. J. Andersen; F. J. R. Taylor; P. J. Harrison; A. D. Cembella
This paper describes a model for assessment of coastal and offshore shellfish aquaculture at the farm-scale. The Farm Aquaculture Resource Management (FARM) model is directed both at the farmer and the regulator, and has three main uses: (i) prospective analyses of culture location and species selection; (ii) ecological and economic optimisation of culture practice, such as timing and sizes for
J. G. Ferreira; A. J. S. Hawkins; S. B. Bricker
More than 20 countries have either established or proposed regulatory limits for one or more of the paralytic shellfish poisoning (PSP) toxins as they occur in seafood products. PSP toxin levels are generally estimated using the standard AOAC mouse bioassay, yet because of various limitations of this method [e.g. high variability (± 20%), low sensitivity, limited sample throughput and use
Gregory J. Doucette; Margaret M. Logan; John S. Ramsdell; Frances M. Van Dolah
Solid-phase adsorption toxin tracking (SPATT) technology was developed as an effective passive sampling method for dissolved diarrhetic shellfish poisoning (DSP) toxins in seawater. HP20 and SP700 resins have been reported as preferred adsorption substrates for lipophilic algal toxins and are recommended for use in SPATT testing. However, information on the mechanism of passive adsorption by these polymeric resins is still
Aifeng Li; Feifei Ma; Xiuli Song; Rencheng Yu
The diarrhetic shellfish poisoning toxins okadaic acid (OA) and dinophysistoxin-1 (DTX-1) are potent phosphatase inhibitors produced by certain species of marine dinoflagellates. OA can cause hyperphosphorylation of a broad range of animal and higherpalnt proteins, but little is known regarding the effects of the DSP toxins on marine organisms or their biological function. A variety of microalgae, including a clone
A. J. Windust; J. L. C. Wright; J. L. McLachlan
The authors determined that the organic sulfur compounds usually contained in crude oil can be used as a marker of oil pollution in shellfish and fish. Short-necked clams and eels were maintained in a controlled laboratory environment in water with suspension of crude oil. Mass spectra and mass chromatograms of short-necked clam extract showed the presence of organic sulfur compounds.
M. Ogata; Y. Miyake
In order to evaluate the main source of nutrients fueling primary production in a shellfish farming bay (Gamak Bay) in the southern sea of Korea, we measured the concentrations of nutrients and radium isotopes in stream water, surface seawater, and coastal groundwater in May 2006, August 2006, and November 2007. Using a 226Ra mass balance model, submarine groundwater discharge (SGD)
Dong-Woon Hwang; Guebuem Kim; Won-Chan Lee; Hyun-Taik Oh
Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration
M.-C. Choi; D. P. H. Hsieh; P. K. S. Lam; W.-X. Wang
COASTAL *FISH is a computerized database that includes current information on spawning, nursery areas, migrations, distribution, and environmental requirements of finfish and shellfish. This User's Guide was prepared to access and query the finfish and shell fish database. It provides basic information required to query the database.
Jones, B.; Benson, N.
Volety, Aswani K., S.G. Tolley and James T. Winstead. 2002. Utilizing Shellfish Responses to Set Target Water Quality Conditions for the Restoration of Oyster Reefs in the Caloosahatchee Estuary, Florida. Presented at the International Workshop on Restoration of Benthic Invertebr...
The Gulf of Maine experiences annual closures of shellfish harvesting due to the accumulation of toxins produced by dinoflagellates of the genus Alexandrium. Factors controlling the timing, location, and magnitude of these events in eastern Maine remain poorly understood. Previous work identified possible linkages between interannual variability of oceanographic variables and shellfish toxicity along the western Maine coastline but no such linkages were evident along the eastern Maine coast in the vicinity of Cobscook Bay, where strong tidal mixing tends to reduce seasonal variability in oceanographic properties. Using 21 years (1985-2005) of shellfish toxicity data, interannual variability in two metrics of annual toxicity, maximum magnitude and total annual toxicity, from stations in the Cobscook Bay region are examined for relationships to a suite of available environmental variables. Consistent with earlier work, no (or only weak) correlations were found between toxicity and oceanographic variables, even those very proximate to the stations such as local sea surface temperature. Similarly no correlations were evident between toxicity and air temperature, precipitation or relative humidity. The data suggest possible connections to local river discharge, but plausible mechanisms are not obvious. Correlations between toxicity and two variables indicative of local meteorological conditions, dew point and atmospheric pressure, both suggest a link between increased toxicity in these eastern Maine stations and weather conditions characterized by clearer skies/drier air (or less stormy/humid conditions). As no correlation of opposite sign was evident between toxicity and local precipitation, one plausible link is through light availability and its positive impact on phytoplankton production in this persistently foggy section of coast. These preliminary findings point to both the value of maintaining long-term shellfish toxicity sampling and a need for inclusion of weather variability in future modeling studies aimed at development of a more mechanistic understanding of factors controlling interannual differences in eastern Gulf of Maine shellfish toxicity.
Horecka, Hannah M.; Thomas, Andrew C.; Weatherbee, Ryan A.
The relationship between toxic marine microalgae species and climate change has become a high profile and well discussed topic in recent years, with research focusing on the possible future impacts of changing hydrological conditions on Harmful Algal Bloom (HAB) species around the world. However, there is very little literature concerning the epidemiology of these species on marine organisms and human health. Here, we examine the current state of toxic microalgae species around the UK, in two ways: first we describe the key toxic syndromes and gather together the disparate reported data on their epidemiology from UK records and monitoring procedures. Secondly, using NHS hospital admissions and GP records from Wales, we attempt to quantify the incidence of shellfish poisoning from an independent source. We show that within the UK, outbreaks of shellfish poisoning are rare but occurring on a yearly basis in different regions and affecting a diverse range of molluscan shellfish and other marine organisms. We also show that the abundance of a species does not necessarily correlate to the rate of toxic events. Based on routine hospital records, the numbers of shellfish poisonings in the UK are very low, but the identification of the toxin involved, or even a confirmation of a poisoning event is extremely difficult to diagnose. An effective shellfish monitoring system, which shuts down aquaculture sites when toxins exceed regularity limits, has clearly prevented serious impact to human health, and remains the only viable means of monitoring the potential threat to human health. However, the closure of these sites has an adverse economic impact, and the monitoring system does not include all toxic plankton. The possible geographic spreading of toxic microalgae species is therefore a concern, as warmer waters in the Atlantic could suit several species with southern biogeographical affinities enabling them to occupy the coastal regions of the UK, but which are not yet monitored or considered to be detrimental. PMID:21645342
Hinder, Stephanie L; Hays, Graeme C; Brooks, Caroline J; Davies, Angharad P; Edwards, Martin; Walne, Anthony W; Gravenor, Mike B
The relationship between toxic marine microalgae species and climate change has become a high profile and well discussed topic in recent years, with research focusing on the possible future impacts of changing hydrological conditions on Harmful Algal Bloom (HAB) species around the world. However, there is very little literature concerning the epidemiology of these species on marine organisms and human health. Here, we examine the current state of toxic microalgae species around the UK, in two ways: first we describe the key toxic syndromes and gather together the disparate reported data on their epidemiology from UK records and monitoring procedures. Secondly, using NHS hospital admissions and GP records from Wales, we attempt to quantify the incidence of shellfish poisoning from an independent source. We show that within the UK, outbreaks of shellfish poisoning are rare but occurring on a yearly basis in different regions and affecting a diverse range of molluscan shellfish and other marine organisms. We also show that the abundance of a species does not necessarily correlate to the rate of toxic events. Based on routine hospital records, the numbers of shellfish poisonings in the UK are very low, but the identification of the toxin involved, or even a confirmation of a poisoning event is extremely difficult to diagnose. An effective shellfish monitoring system, which shuts down aquaculture sites when toxins exceed regularity limits, has clearly prevented serious impact to human health, and remains the only viable means of monitoring the potential threat to human health. However, the closure of these sites has an adverse economic impact, and the monitoring system does not include all toxic plankton. The possible geographic spreading of toxic microalgae species is therefore a concern, as warmer waters in the Atlantic could suit several species with southern biogeographical affinities enabling them to occupy the coastal regions of the UK, but which are not yet monitored or considered to be detrimental.
The biosensor consisted of a sodium electrode and covered with the frog bladder membrane within a flow cell was tested for the estimation of tetrodotoxin (TTX) and saxitoxin (STX). This sensor was applied to detect very low amounts of the Na+ channel blockers, STX and TTX, in different shellfishes and swellfishes. A good agreement was obtained between TTX activities determined by mouse assay and amounts of Na+ channel blockers estimated by frog membrane sensor. The lowest level of TTX (fg) that can be determined by frog membrane sensor does not cause human poisoning. The channel blockers in short-necked clam, which was assumed to be STX, were monitored by this sensor continuously every week for one year. It was discovered that the STX content increased from July until September and then decreased from October until March. The biosensor proposed here may be used for the estimation of STX and TTX conventionally in the future. PMID:9678185
Cheun, B S; Takagi, S; Hayashi, T; Nagashima, Y; Watanabe, E
In this study, a total of 3432 aquatic animals covering eight species were tested by specific polymerase chain reaction (PCR) assays to investigate the presence of Toxoplasma gondii, in China. The results showed that, out of 618 Procambarus clarkii samples collected from the different areas in southeast China, four samples from Jiangxi province were positive. Of 456 Hypophthalmichthys molitrix samples, one sample from Jiangsu province was positive. In addition, there was only one positive sample collected from Shandong province out of 813 Macrobranchium nipponense samples. All other samples, including 309 Cyprinus carpio, 398 Concha Ostreae, 98 Momopterus albus, 426 Penaeus monodon Fabricius and 309 Carassius auratus were negative. The results suggested that the level of T. gondii in aquatic animals was low in China. However, the importance of T. gondii in aquatic animals should not be ignored. Consumption of contaminated raw shellfish may represent a considerable health threat. PMID:24331298
Zhang, Meng; Yang, Zhen; Wang, Shuai; Tao, LongFei; Xu, LiXin; Yan, RuoFeng; Song, XiaoKai; Li, XianRrui
Purple clams (Hiatula rostrata Lighttoot) accumulated paralytic shellfish poisoning (PSP) toxins produced by a toxic strain of the dinoflagellate Alexandrium minutum Halim for subsequent study of toxin distribution during depuration (detoxification by a nontoxic microalgal diet or starvation). The results confirm the data in the literature concerning the high toxicity of the digestive gland, and the depuration efficiency between feed with nontoxic microalgae and starvation is similar. The toxin profile of the purple clams was similar with that of Alexandrium minutum at the end of the exposure period; GTX4 and GTX1 were dominant. However, at the end of the depuration period, GTX3 and GTX2 were dominant. The non-visceral tissues were toxic after feeding with toxic algae. The toxicity was low and the profile were also similar with that of the toxic algae. No PSP toxins other than GTX-1, 2, 3 and 4 were detected in the experimental period. PMID:11223092
Chen, C Y; Chou, H N
During the monitoring programme of harmful algal blooms established along the south Atlantic coast of Morocco, a bimonthly determination of harmful algae and phycotoxins analysis in Perna perna was carried out from May 2003 to December 2004. Results of mouse bioassay (in organs and whole flesh) showed a seasonal evolution of paralytic shellfish poisoning (PSP) toxin. The mussel's contamination was associated with the occurrence in water of Alexandrium minutum. The PSP toxin profile obtained with high-performance liquid chromatography (HPLC/FD) revealed the dominance of gonyautoxins GTX2 and GTX3 and a minority of GTX1, GTX4 and saxitoxin (STX). This profile explains that the toxicity was mainly associated with A. minutum. PMID:18237757
Abouabdellah, Rachid; Taleb, Hamid; Bennouna, Asmae; Erler, Katrin; Chafik, Abdeghani; Moukrim, Abdelatif
Purple clams (Hiatula diphos Linnaeus) accumulate paralytic shellfish poisoning (PSP) toxins produced by a toxic strain of the dinoflagellate Alexandrium minutum Halim in a laboratory study. The maximal toxicity of PSP toxins attained 31.3m MU/g after 20 days exposure. The toxin profile of H. diphos was similar to that reported for A. minutum at the end of the exposure period; and GTX1 was dominant. GTX congeners were found in muscle on day 16 and day 20, these substances could be detected during the depuration period as well. GTX1 was detected in the siphon only on day 32. The results show that H. diphos accumulates PSP toxins according to the amount and toxin profile of ingested A. minutum. PMID:16137734
Chou, Hong Nong; Huang, Chen Ping; Chen, Chih Yu
This report defines the role of nearshore ecosystems, such as wetlands and seagrass meadows, as nurseries for populations of fish and shellfish that may be of commercial value to humans. The ecological value of nursery habitats in relation to the life cycle of many species of fish and invertebrates is mentioned as the nursery-role hypothesis is discussed. Biological, physical, chemical, and landscape factors that contribute to site-specific variation of nurseries are listed. Implications for research, conservation, management and restoration are listed, while highlighting key threats to coastal ecosystems. Issues in Ecology is an ongoing series of reports designed to present major ecological issues in an easy-to-read manner. This Issue summarizes the consensus of a panel of scientific experts based on the information that was current and available at the time of its publication in 2003.
The occurrence of brominated dioxins (PBDD/Fs) and polybrominated diphenyl ethers (PBDEs) was investigated in commonly consumed species of marine shellfish in the UK. Individual samples of Pacific oysters (Crassostrea gigas), native oysters (Ostrea edulis), mussels (Mytilus edulis), scallops (Pecten maximus), and cockles (Cerastoderma edule) were collected from different coastal regions between 2006 and 2007. Samples of a particular species from each site were composited and 60 samples were analysed. Polybrominated dibenzofurans (PBDFs) occurred more frequently and generally at a higher level than polybrominated dibenzodioxins (PBDDs), except for 237-TriBDD, which was the predominant PBDD/F congener in some species, notably oysters. This profile may reflect the environmental distribution of these compounds and the effects of removal mechanisms, such as degradation, selective uptake and metabolism. PBDEs were detected in all samples. The dominant congeners were BDEs 47, 49, 99 and 100 and, to a lesser extent, BDEs 66 and 154. The occurrence of BDE-209 was observed in most samples and appears to be species selective, with the highest values occurring almost exclusively in mussels and cockles. Among the species studied, oysters and mussels displayed relatively higher levels of both sets of contaminants; native oysters, in particular, showed elevated levels of 237-TriBDD (up to 14.5 ng/kg). In general, contaminant levels appeared to be consistent with the extent of local industrialisation with lower levels observed in more remote areas such as the north of Scotland. Polybrominated biphenyls (PBBs) were also measured, and PBBs 49, 52 and 77 were the most frequently detected, although levels were very low. Dietary intakes, estimated for PBDD/Fs, showed that 237-TriBDD from single portions of oysters constituted a high proportion of the total dietary intake of the congener but, otherwise, dietary intakes of PBDD/Fs from shellfish were relatively low. PMID:19680967
Fernandes, Alwyn; Mortimer, David; Gem, Martin; Dicks, Pamela; Smith, Frankie; White, Shaun; Rose, Martin
Background Despite Noroviruses (NV, previously "Norwalk-like viruses") being a leading cause of acute gastroenteritis outbreaks, the impact of NV infection is at present unknown and little information is available about strains circulating in Italy. In April 2002 an outbreak of gastroenteritis occurred in the province of Bari (South-east Italy), involving several households. Methods A retrospective cohort study was performed in order to assess risk factors associated with illness. All households where a case occurred were included in the study. Faecal specimens were collected from ill individuals. NV-specific RT-PCR was performed. Eleven samples of mussels were collected from fish-markets involved in the outbreak. A nested PCR was used for mussel samples. Results One hundred and three cases, detected by means of active surveillance, met the case definition. Raw shellfish eating was the principal risk factor for the disease, as indicated by the analytic issues (Risk Ratio: 1.50; IC 95%: 1.18 – 1.89; p < 0.001). NVs were found by means of RT-PCR of all the stool specimens from the 24 patients tested. Eleven samples of shellfish from local markets were tested for the presence or NVs; six were positive by nested PCR and genotypes were related to that found in patients' stools. Conclusion This is the first community outbreak caused by NVs related to sea-food consumption described in Italy. The study confirms that the present standards for human faecal contamination do not seem to be a reliable indicator of viral contaminants in mussels.
Prato, Rosa; Lopalco, Pier Luigi; Chironna, Maria; Barbuti, Giovanna; Germinario, Cinzia; Quarto, Michele
For the supply of fish and shellfish to consumers in fresh condition, clean handling after catch from the sea is essential.\\u000a According to HACCP (Hazard Analysis and Critical Control Points), it is important to meet such requirement by keeping fish\\u000a and shellfish under a certain low temperature and clean conditions after catching. The deep ocean water (DOW) characterized\\u000a by low
Masayuki Mac Takahashit; Kazunori Yamashita
Most-Probable-Number Loop-Mediated Isothermal Amplification-Based Procedure Enhanced with K Antigen-Specific Immunomagnetic Separation for Quantifying tdh(+) Vibrio parahaemolyticus in Molluscan Shellfish.
Although thermostable direct hemolysin-producing (tdh(+)) Vibrio parahaemolyticus is the leading cause of seafood-borne gastroenteritis, the enumeration of tdh(+) V. parahaemolyticus remains challenging due to its low densities in the environment. In this study, we developed a most-probable-number (MPN)-based procedure designated A-IS(1)-LAMP, in which an immunomagnetic separation (IMS) technique targeting as many as 69 established K antigens and a loop-mediated isothermal amplification (LAMP) assay targeting the thermostable direct hemolysin (tdh) gene were applied in an MPN format. Our IMS employed PickPen, an eight-channel intrasolution magnetic particle separation device, which enabled a straightforward microtiter plate-based IMS procedure (designated as PickPen-IMS). The ability of the procedure to quantify a wide range of tdh(+) V. parahaemolyticus levels was evaluated by testing shellfish samples in Japan and southern Thailand, where shellfish products are known to contain relatively low and high levels of total V. parahaemolyticus, respectively. The Japanese and Thai shellfish samples showed, respectively, relatively low (< 3 to 11 MPN/10 g) and considerably higher (930 to 110,000 MPN/10 g) levels of tdh(+) V. parahaemolyticus, raising concern about the safety of Thai shellfish products sold to domestic consumers at local morning markets. LAMP showed similar or higher performance than conventional PCR in the detection and quantification of a wide range of tdh(+) V. parahaemolyticus levels in shellfish products. Whereas a positive effect of PickPen-IMS was not observed in MPN determination, PickPen-IMS was able to concentrate tdh(+) V. parahaemolyticus 32-fold on average from the Japanese shellfish samples at an individual tube level, suggesting a possibility of using PickPen-IMS as an optional tool for specific shellfish samples. The A-IS(1)-LAMP procedure can be used by any health authority in the world to measure the tdh(+) V. parahaemolyticus levels in shellfish products. PMID:24988012
Tanaka, Natsuko; Iwade, Yoshito; Yamazaki, Wataru; Gondaira, Fumio; Vuddhakul, Varaporn; Nakaguchi, Yoshitsugu; Nishibuchi, Mitsuaki
Climate change is expected to affect food and feed safety, including the occurrence of natural toxins in primary crop and seafood production; however, to date, quantitative estimates are scarce. This study aimed to estimate the impact of climate change effects on mycotoxin contamination of cereal grains cultivated in the terrestrial area of north west Europe, and on the frequency of harmful algal blooms and contamination of shellfish with marine biotoxins in the North Sea coastal zone. The study focused on contamination of wheat with deoxynivalenol, and on abundance of Dinophysis spp. and the possible relationship with diarrhetic shellfish toxins. The study used currently available data and models. Global and regional climate models were combined with models of crop phenology, mycotoxin prediction models, hydrodynamic models and ecological models, with the output of one model being used as input for the other. In addition, statistical data analyses using existing national datasets from the study area were performed to obtain information on the relationships between Dinophysis spp. cell counts and contamination of shellfish with diarrhetic shellfish toxins as well as on frequency of cereal cropping. In this paper, a summary of the study is presented, and overall conclusions and recommendations are given. Climate change projections for the years 2031-2050 were used as the starting point of the analyses relative to a preceding 20-year baseline period from which the climate change signal was calculated. Results showed that, in general, climate change effects lead to advanced flowering and harvest of wheat, and increased risk of contamination of wheat with deoxynivalenol. Blooms of dinoflagellates were estimated to occur more often. If the group of Dinophysis spp. behaves similarly to other flagellates in the future then frequency of harmful algal blooms of Dinophysis spp. may also increase, but consequences for contamination of shellfish with diarrhetic shellfish toxins are uncertain. Climate change will also have indirect effects on toxin contamination, which may be equally important. For example, the frequency of cropping of wheat and maize in north Europe was projected to increase under climate change, which will also increase the risk of contamination of the grains with deoxynivalenol. Risk managers are encouraged to consider the entire range of the predictions of climate change effects on food safety hazards, rather than median or average values only. Furthermore, it is recommended to closely monitor levels of mycotoxins and marine biotoxins in the future, in particular related to risky situations associated with favourable climatic conditions for toxin producing organisms. In particular, it is important to pay attention to the continuity of collecting the right data, and the availability and accessibility of databases. On a European level, it is important to stress the need for harmonisation of terminology and data collection. PMID:22891967
van der Fels-Klerx, H J; Olesen, J E; Naustvoll, L-J; Friocourt, Y; Mengelers, M J B; Christensen, J H
Mytilid bivalves employ a set of threads (the byssus) to attach themselves to both hard and soft substrates. In this study, we measured the mechanical properties of byssal threads from two semi-infaunal mytilids (Geukensia demissa Dillwyn and Modiolus modiolus Linnaeus) and two epifaunal mytilids (Mytilus californianus Conrad and Mytilus edulis Linnaeus). We compared material properties with and without the assumption that changes of length and area during tensile testing are insignificant, demonstrating that previous researchers have overestimated extensibility values by 30% and may also have underestimated strength values. We detected significant differences in thread properties among tested mytilid species, contrary to previous findings. Threads from semi-infaunal species were significantly thinner than those from epifaunal species, perhaps to allow the production of a greater number of threads, which form a dense network within the substrate. Geukensia demissa threads were weaker than those of the other species, and had a significantly lower stiffness at failure. Modiolus modiolus threads were significantly stiffer than M. edulis threads but also significantly less extensible, suggesting a trade-off between stiffness and extensibility. The only thread property that did not show significant differences across species was toughness - even when byssal threads differ in strength or stiffness, they seem to absorb similar amounts of energy per unit volume prior to failure. This study reveals notable differences between the byssal thread properties of different mytilid bivalves and provides a reliable and thorough methodology for future comparative studies. PMID:19411537
Pearce, Trevor; Labarbera, Michael
The authors determined that the organic sulfur compounds usually contained in crude oil can be used as a marker of oil pollution in shellfish and fish. Short-necked clams and eels were maintained in a controlled laboratory environment in water with suspension of crude oil. Mass spectra and mass chromatograms of short-necked clam extract showed the presence of organic sulfur compounds. Capillary column gas chromatography-mass chromatograms of crude oil and extract from the soft body of a short-necked clam showed the presence of organic sulfur compounds. Besides sulfur components, various other compounds were contained in crude oil and short-necked clam. Mass chromatograms of crude oil and the extract from eel flesh showed the presence of alkyl benzothiophene, dibenzothiophene, naphthalene, and alkyl naphthalene. Data indicated that the organic sulfur compounds and polyaromatic compounds could serve as markers of oil pollution in shellfish and fish.
Ogata, M.; Miyake, Y.
Azaspiracids (AZA) are polyether marine toxins that accumulate in various shellfish species and have been associated with severe gastrointestinal human intoxications since 1995. This toxin class has since been reported from several countries, including Morocco and much of western Europe. A regulatory limit of 160 ?g AZA/kg whole shellfish flesh was established by the EU in order to protect human health; however, in some cases, AZA concentrations far exceed the action level. Herein we discuss recent advances on the chemistry of various AZA analogs, review the ecology of AZAs, including the putative progenitor algal species, collectively interpret the in vitro and in vivo data on the toxicology of AZAs relating to human health issues, and outline the European legislature associated with AZAs.
Twiner, Michael J.; Rehmann, Nils; Hess, Philipp; Doucette, Gregory J.
Purple clams (Hiatula rostrata Lighttoot) accumulate paralytic shellfish poisoning (PSP) toxins produced by a toxic strain of the dinoflagellate Alexandriun minutum Halim. The results confirm the data of our previous study concerning the muscle and siphon that were not showing a gradual rise in toxicity when shellfish accumulated more A. minutum. However, muscle and siphon are intermittently toxic both in exposure and depuration period in laboratory cultured purple clams. PSP toxins were detected in outdoor cultured purple clams, whereas no A. minutum were found in the culture pond during most of the survey time. The outdoor cultured purple clams need longer time to decrease toxicity to allowable levels than laboratory cultured purple clams. It was shown that laboratory data may not predict times over which pond-cultured purple clams may prove toxic to consumers. PMID:12269475
Chen, Chih Yu; Chou, Hong Nong
Diarrhetic shellfish poisoning (DSP) is a severe gastrointestinal illness caused by consumption of shellfish contaminated with DSP toxins that are originally produced by toxic dinoflagellates. Based on their structures, DSP toxins were initially classified into three groups, okadaic acid (OA)/dinophysistoxin (DTX) analogues, pectenotoxins (PTXs), and yessotoxins (YTXs). Because PTXs and YTXs have been subsequently shown to have no diarrhetic activities, PTXs and YTXs have recently been eliminated from the definition of DSP toxins. Mouse bioassay (MBA), which is the official testing method of DSP in Japan and many countries, also detects PTXs and YTXs, and thus alternative testing methods detecting only OA/DTX analogues are required in DSP monitoring. Electrospray ionization (ESI) liquid chromatography-mass spectrometry (LC-MS) is a very powerful tool for the detection, identification and quantification of DSP and other lipophilic toxins. In the present review, application of ESI LC-MS techniques to the analysis of each toxin group is described. PMID:21666353
Suzuki, Toshiyuki; Quilliam, Michael A
Three assays were developed to enumerate total and fecal Escherichia coli and total Vibrionaceae in shellfish, seawater, and other foods and environmental samples. Assays involve membrane overlays of overnight colonies on nonselective agar plates to detect ?-glucuronidase and lysyl aminopeptidase activities for E. coli and Vibrionaceae, respectively. Cellulose membranes containing the substrates 4-methylumbeferyl-?-D-glucuronide (MUG) produced a bright blue fluorescence when overlaid onto E. coli, while L-lysyl-7-amino-4-trifluoromethylcoumarin produced green fluorescent foci when overlaid onto Vibrionaceae family members. A multiplex assay was also developed for simultaneously enumerating total E. coli and total Vibrionaceae in oysters and seawater. Overall, 65% of overlaid E. coli (non-O157:H7) were MUG-positive, compared with 62% as determined by the most-probable-number-MUG assay. The overlays are rapid, simple, and cost effective for quantification purposes. This research provides practical alternatives for monitoring bacterial indicators and potential pathogens in complex samples, including molluscan shellfish.
Richards, Gary P.; Watson, Michael A.
Contamination of shellfish with diarrhetic shellfish poisoning (DSP) toxins readily occurs during algal blooms. Such phenomena raise important public health concerns and thus comprise a constant challenge to shellfish farmers, the seafood industry and health services, considering the increasing occurrence of toxic episodes around the world. To avoid the detrimental effects of such episodes, research has focused on the use of various detoxification methodologies that should be rapid, efficient, easy to apply, and will not alter the quality and sensory properties of shellfish. In the present study, both ozonation (15 mg kg(-1) for 6 h) and ?-irradiation (6 kGy) were utilised in order to reduce the toxin content of contaminated shucked mussels, collected during the DSP episodes of 2007 and 2009 in Greece. DSP toxicity was monitored using the mouse bioassay (MBA) whilst the determination of toxin content of the okadaic acid (OA) group (both free and esterified forms) was carried out by LC/MS/MS analysis. Toxin reduction using ?-irradiation was in the range of 12-36%, 8-53% and 10-41% for free OA, OA esters and total OA, respectively. The appearance and texture of irradiated mussels deteriorated, pointing to a low potential for commercial use of this method. Ozonation of mussels resulted in toxin reduction in the range of 6-100%, 25-83% and 21-66% for free OA, OA esters and total OA, respectively. Reduction of OA content was substantially higher in homogenised mussel tissue compared with that of whole shucked mussels. In addition, differences detected with regard to quality parameters (TBA, sensory attributes) between ozonated and control mussels were not considerable. Even though varying percentage reductions in OA and its derivatives were achieved using ozonation under specific experimental conditions tested, it is postulated that upon optimisation ozonation may have the potential for post-harvest commercial DSP detoxification of shucked mussels. PMID:21985774
Louppis, A P; Katikou, P; Georgantelis, D; Badeka, A V; Kontominas, M G
Liquid chromatography (LC)-linked protein phosphatase 1/2A (PP-1/PP2A) bioassay was used to quantitatively identify diarrhetic shellfish toxins in marine phytoplankton (cultured and natural assemblages) and commercially available mussels. Using this approach, multiple protein phosphatase inhibitor profiles of varying composition were found in diarrhetic mussels from Holland and Canada. Based on LC elution positions and relative activity versus PP-1 and PP-2A, at least six inhibitors distinct from known diarrhetic shellfish toxins were identified and termed mussel phosphatase inhibitor (MPI) 19,22,23,25,33 and 42. The levels of these inhibitors, in okadaic acid equivalent units, varied from 100 pg to 3350 ng per g shellfish tissue. The combined levels of PP-1/2A inhibitors in all instances superseded that of okadaic acid/dinophysistoxin-1 and may contribute to the diarrhetic shellfish toxin profile of the contaminated mussels. The efficacy of LC-protein phosphatase bioassay was established for cultured phytoplankton where picogram levels of okadaic acid could be detected from microgram extracts of Prorocentrum lima. Analyses of plankton net tows from estuarine mussel culture sites in Eastern Canada revealed a heterogeneous population of protein phosphatase inhibitors, with dinophysistoxin-1 being most prevalent. This toxin was predominant for at least 2 months in mussel populations in the immediate vicinity of plankton sampling sites. The results are consistent with a hypothetical model in which marine bacteria, cyanobacteria and dinoflagellates combine to produce a variety of protein phosphatase inhibitors effective against signal transduction pathways in higher eukaryotes. PMID:8383366
Luu, H A; Chen, D Z; Magoon, J; Worms, J; Smith, J; Holmes, C F
Sediment and water samples were collected for Alexandrium fundyense spatial and temporal distribution and abundance at more than 120 locations throughout the Bay of Fundy during the summers and winters of 1980-1984. These broad surveys have been repeated at various times through the past 30 years, with more regular sampling since 2004. In addition, A. fundyense abundance has been monitored at several locations within the Bay of Fundy at weekly intervals from April to November and monthly during the remaining months since 1988. Paralytic Shellfish Poisoning (PSP) toxins in shellfish (notably Mya arenaria) have also been monitored at multiple locations in the Bay of Fundy since 1943. The datasets were examined to determine relationships and roles between overwintering resting cysts, bloom initiation, bloom decline, motile cell dispersal and A. fundyense motile populations and resulting shellfish toxicity since 1980. Cysts are widely dispersed throughout the Bay of Fundy in the offshore, inshore and intertidal zones with the largest deposits located in the offshore in silt/clay sediments to the east and north of Grand Manan Island at depths of 60-180 m. Results show that there is a constant stable source of cysts in the Bay of Fundy with highest concentrations of cysts (9780 cysts cm-3) observed in 2010 and highest concentrations of A. fundyense motile cells (18×106 cells L-1) observed in 1980. Interannual changes in abundance in A. fundyense populations, resting cysts and the temporal trends in M. arenaria toxicity are discussed. Results show that there was no relationship between the abundance of overwintering cysts and the magnitude of A. fundyense blooms. The offshore seed beds appear to be relatively constant in cyst density among most years and serve as an important source for the motile cells that lead to initiation of major blooms and resulting shellfish toxicity throughout the Bay of Fundy.
Martin, Jennifer L.; LeGresley, Murielle M.; Hanke, Alex R.
A study was conducted to assess the feasibility for the production of sterile, stable and homogenous shellfish reference materials containing known concentrations of paralytic shellfish poisoning (PSP) toxins. Pacific oysters were contaminated with toxins following mass culturing of toxic algae and shellfish feeding experiments. Live oysters were shucked and tissues homogenised, before measuring into multiple aliquots, with one batch subjected to gamma irradiation treatment and the other remaining untreated. The homogeneity of both batches of samples was assessed using a pre-column oxidation liquid chromatography with fluorescence detection (Pre-COX LC-FLD) method and shown to be within the limits of normal within-batch repeatability. A twelve-month stability experiment was conducted for both untreated and gamma irradiated batches, specifically examining the effects of long term storage at -20 °C, +4 °C and +40 °C. Results indicated mostly good stability of PSP toxins in both materials when stored frozen at -20 °C, but with the instability of GTX2&3 concentrations in the untreated tissues eliminated in the irradiated tissues. Analysis using a post-column oxidation (PCOX) LC-FLD method also showed epimerisation in both GTX1&4 and GTX2&3 epimeric pairs in untreated samples after only 6 months frozen storage. This issue was not present in the tissues irradiated before long term storage. Biological activity testing confirmed the absence of bacteria in the irradiated samples throughout the 12 month study period. With such results there was clear evidence for the potential of increasing the scale of the mass culturing and shellfish feeding for the production of large batches of tissue suitable for the preparation of a certified matrix reference material. Overall results demonstrated the feasibility for production of oyster reference materials for PSTs, with evidence for prolonged stability following gamma irradiation treatment and storage at -20 °C. PMID:23796726
Turner, Andrew D; Lewis, Adam M; Hatfield, Robert G; Powell, Andy L; Higman, Wendy A
An isoform of the paralytic shellfish poison (PSP)-specific receptor saxiphilin, from the tropical centipedeEthmostigmus rubripes,was used as the basis for a radiometric, high-throughput, microtiter plate assay for this group of toxins. Characterization of the assay revealed that it was able to detect several representatives from the various structural PSP subgroups and yet was insensitive toward tetrodotoxin. To test the utility
Lyndon E. Llewellyn; Jason Doyle; Andrew P. Negri
The production and composition of the paralytic shellfish toxins (PSTs) of 2 Alexandrium tamarense strains (CI01 and HK9301) in the presence and absence of bacteria were investigated. Attempts were made to produce bacteria-free dinoflagellate cultures using 2 antibiotic treatments. Antibiotic treatment 1 (penicillin-G and streptomycin) partially killed bacteria in 2 dinoflagellate cultures, while antibiotic treatment 2 (penicillin-G, streptomycin, ciprofloxacin, and
Alvin Y. T. Ho; Dennis P. H. Hsieh; Pei-Yuan Qian
While there is increasing evidence that marine bacteria are involved in the production of paralytic shellfish toxins in algal\\u000a blooms, the exact roles of the bacteria and microalgae have proved elusive. A novel experimental approach to this problem\\u000a involved incubating parallel cultures of toxin producing Alexandrium minutum Anokoha A in the dark and in a natural daylight cycle. High-performance liquid
Elizabeth W. Maas; Heather Joan Linton Brooks
The presence of paralytic shellfish poisoning (PSP), diarrheic shellfish poisoning (DSP), and amnesic shellfish poisoning (ASP) toxins in seafood is a severe and growing threat to human health. In order to minimize the risks of human exposure, the maximum content of these toxins in seafood has been limited by legal regulations worldwide. The regulated limits are established in equivalents of the main representatives of the groups: saxitoxin (STX), okadaic acid (OA), and domoic acid (DA), for PSP, DSP, and ASP, respectively. In this study a multidetection method to screen shellfish samples for the presence of these toxins simultaneously was developed. Multiplexing was achieved using a solid-phase microsphere assay coupled to flow-fluorimetry detection, based on the Luminex xMap technology. The multidetection method consists of three simultaneous competition immunoassays. Free toxins in solution compete with STX, OA, or DA immobilized on the surface of three different classes of microspheres for binding to specific monoclonal antibodies. The IC50 obtained in the buffer was similar in single- and multidetection: 5.6 ± 1.1 ng/mL for STX, 1.1 ± 0.03 ng/mL for OA, and 1.9 ± 0.1 ng/mL for DA. The sample preparation protocol was optimized for the simultaneous extraction of STX, OA, and DA with a mixture of methanol and acetate buffer. The three immunoassays performed well with mussel and scallop matrixes displaying adequate dynamic ranges and recovery rates (around 90% for STX, 80% for OA, and 100% for DA). This microsphere-based multidetection immunoassay provides an easy and rapid screening method capable of detecting simultaneously in the same sample three regulated groups of marine toxins. PMID:23859142
Fraga, María; Vilariño, Natalia; Louzao, M Carmen; Rodríguez, Paula; Campbell, Katrina; Elliott, Christopher T; Botana, Luis M
The Australian fresh-water mussel Alathyria condola accumulated high levels of paralytic shellfish poisoning (PSP) toxins when fed the neurotoxic cyanobacterium Anabaena circinalis, shown recently to contain high concentrations of C-toxins and gonyautoxins. Significant accumulation (>; 80 ?g\\/100 g of mussel flesh) was detected following 2–3 days exposure to water containing 2 × 105 cells\\/mlA. circinalis. Only trace accumulation of PSP
Andrew P. Negri; Gary J. Jones
Headspace solid phase micro-extraction (HS-SPME) has been applied as a sampling technique for the determination of petroleum contamination in shellfish using capillary gas chromatography-mass spectrometry (GC-MS). A poly(dimethylsiloxane) fused silica fibre (100 microm thickness) was found to be satisfactory for the extraction of a range of aliphatic hydrocarbons (HCs) from homogenised shellfish tissues. The SPME conditions, including temperature, salt content, extraction time and desorption temperature, were optimised for a range of aliphatic HCs (C9-C20). A methyl silicone column GC (12 m x 0.20 mm, 0.33 microm layer thickness) was used with a temperature programme from 40 to 260 degrees C and the HCs were determined within a mass range of m/ z=50-550 in electron impact mode. Calibration range was from 10 to 5000 ng/g with linear correlation coefficients ( r(2)) of 0.982 for nonane to 0.997 for octadecane. Detection limits for aliphatic HCs, spiked into shellfish (mussel) tissues, varied from 3.6 ng/g (tetradecane) to 51 ng/g (eicosane) and relative standard deviation (% RSD) values ranged from 1.4% (hexadecane) to 24.3%(eicosane). PMID:12043016
Stack, Mary A; O'Connell, Sharon; James, Kevin J
This article gives an overview of a range of persistent organic pollutant chemical levels in shellfish (Batissa violacea and Anadara antiquata) species and eel (Gymnothorax flavimarginatus) from Fiji. As there is limited data in published literature to date, this paper reports first data on a range of persistent organic pollutants and highlights the more prominent POP chemicals present in marine biota in Fiji. A significant number of POP chemicals were detected (e.g. 17 PCDD/PCDF, 12dl-PCBs, organochlorine pesticides and brominated flame retardants), the concentrations found were generally low (e.g. parts per billion level). The low levels of contamination are indicative of a low input from long range and short-range transport as well as few local point sources. Also concentrations of POPs in eel and shellfish from Fiji are low in comparison to wild species in other regions and are within acceptable limits for POP chemicals in fish and fishery products set by the European Union. It describes also results of early studies on basic POPs levels in shellfish in several Pacific Island Countries, which generally show relatively low levels. PMID:24568747
Lal, Vincent; Bridgen, Phil; Votadroka, Waisea; Raju, Rupantri; Aalbersberg, William
A lateral flow immunochromatographic (LFIC) test strip based on a colloidal gold-monoclonal antibody (McAb) conjugate was developed for on-site rapid detection of okadaic acid (OA) in shellfish. It applies a competitive format using an immobilized toxin conjugate and free toxin present in samples. The McAb against OA was conjugated with 20-nm colloidal gold as detector reagent. The toxin in the sample competed with the immobilized toxin to bind to the gold conjugated with McAb. The colloidal gold/McAb/toxin mobile complex was not captured by OA-bovine serum albumin (BSA) on the test line, but it was captured by goat anti-mouse immunoglobulin G (IgG) on the control line. The color density of the test line correlated with the concentration of toxin in the range of 10-50 ng ml(-1). The qualitative detection limit of 150 ?g kg(-1) sample was close to the European Union (EU) regulatory limit (160 ?g kg(-1)). Therefore, these strips were able to directly and qualitatively estimate the consuming safety of shellfish. They require no equipment because of available visual results, and they screened numerous samples within 10 min. The results were further confirmed by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). As a food safety screening tool, the test strips are convenient and useful to rapidly on-site test the presence of OA in shellfish products. PMID:22266294
Lu, Shi-Ying; Lin, Chao; Li, Yan-Song; Zhou, Yu; Meng, Xian-Mei; Yu, Shi-Yu; Li, Zhao-Hui; Li, Le; Ren, Hong-Lin; Liu, Zeng-Shan
Forty-eight composite samples of the most commonly consumed fish and shellfish species were prepared from up to 60 individual subsamples of each species and analysed for chlorinated dioxins (PCDD/Fs) and polychlorinated biphenyls (PCBs). These included 24 species of fresh wild fish, seven of farmed fish, seven of fresh shellfish, and ten processed fish and shellfish products. The ISO 17025-accredited analytical methodology used is consistent with the requirements given in European Commission Directive 2002/69/EC. Concentrations ranged from 0.03 ng kg(-1) PCDD/F plus PCB World Health Organization-toxic equivalent quantity (WHO-TEQ) for a sample of surimi, to approximately 6 ng kg(-1) for wild pilchards/sardines. The corresponding range for the ?ICES-6 PCBs was 0.04 ? g kg(-1) to approximately 47 ? g kg(-1). None of the samples showed concentrations above the European Union maximum permitted limits. Averaged PCDD/F and PCB concentrations for the two groups of farmed and wild fish show that there is little difference between the two categories, although individual species may show variations depending on factors such as the sampling location. PMID:24784962
Fernandes, A R; Mortimer, D N; Rose, M; Knowles, T G; White, S; Gem, M
A novel graphene-based voltammetric immunosensor for sensitive detection of okadaic acid (OA) was developed. A simple and efficient electrografting method was utilized to functionalize graphene-modified screen-printed carbon electrodes (GSPE) by the electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt in acidic aqueous solution. Next, the okadaic acid antibody was covalently immobilized on the carboxyphenyl modified graphene electrodes via carbodiimide chemistry. Square wave voltammetry (SWV) was used to investigate the stepwise assembly of the immunosensor. A competitive assay between OA and a fixed concentration of okadaic acid-ovalbumin conjugate (OA-OVA) for the immobilized antibodies was employed for the detection of okadaic acid. The decrease of the [Fe(CN)6]3-/4- reduction peak current in the square wave voltammetry for various concentrations of okadaic acid was used for establishing the calibration curve. A linear relationship between the SWV peak current difference and OA concentration was obtained up to ~5000 ng L-1. The developed immunosensor allowed a detection limit of 19 ng L-1 of OA in PBS buffer. The matrix effect studied with spiked shellfish tissue extracts showed a good percentage of recovery and the method was also validated with certified reference mussel samples.A novel graphene-based voltammetric immunosensor for sensitive detection of okadaic acid (OA) was developed. A simple and efficient electrografting method was utilized to functionalize graphene-modified screen-printed carbon electrodes (GSPE) by the electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt in acidic aqueous solution. Next, the okadaic acid antibody was covalently immobilized on the carboxyphenyl modified graphene electrodes via carbodiimide chemistry. Square wave voltammetry (SWV) was used to investigate the stepwise assembly of the immunosensor. A competitive assay between OA and a fixed concentration of okadaic acid-ovalbumin conjugate (OA-OVA) for the immobilized antibodies was employed for the detection of okadaic acid. The decrease of the [Fe(CN)6]3-/4- reduction peak current in the square wave voltammetry for various concentrations of okadaic acid was used for establishing the calibration curve. A linear relationship between the SWV peak current difference and OA concentration was obtained up to ~5000 ng L-1. The developed immunosensor allowed a detection limit of 19 ng L-1 of OA in PBS buffer. The matrix effect studied with spiked shellfish tissue extracts showed a good percentage of recovery and the method was also validated with certified reference mussel samples. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr32146g
Eissa, Shimaa; Zourob, Mohammed
Runoff from agricultural lands and farm animal feedlots is one of the major sources of fecal coliforms in surface waters, and fecal coliform (FC) bacteria concentrations tend to vary with season because of seasonal variations in climatic factors. However, El Niño -Southern Oscillation (ENSO) events may affect the extent and patterns of seasonality in FC levels in coastal waters. Water quality monitoring data for shellfish management collected during El Niño (1990, 1992, 1997), and La Niña (1999, 2000) years were analyzed to evaluate the extent to which these events influenced Pearl River stage, and bacterial levels, water temperature, and salinity in the western part of Mississippi Sound. Models to predict FC levels in relation to various environmental factors were also developed. In 1990, 1992 and 1997, FC geometric mean counts peaked in late winter (January/February) reaching 120 MPN (February 1990), 165 MPN (January 1992), and 86 MPN (January 1997), and then decreased considerably during spring and summer (1.2 – 19 MPN). Thereafter, FC abundance increased slightly in fall and early winter (1.9 – 24 MPN). Fecal coliform abundance during the 2000 La Niña year was much lower (1.0 – 10.3 MPN) than in 1992 (1.2 – 165 MPN), and showed no seasonal pattern from January to August, perhaps due to the relative scarcity of rainfall in 2000. In 1995 (ENSO neutral year), peak geometric mean FC count (46 MPN) was lower than during El Niño years and occurred in early spring (March). The seasonal and between year variations in FC levels determined the number of days during which the conditionally approved shellfish growing area was opened for harvesting shellfish. For example, from January to April 1997, the area was not opened for shellfish harvesting, whereas in 2000, the number of days during which the area was opened ranged from 6 – 27 (January to April) to 24 – 26 (October to December). ENSO events thus influenced the extent and timing of the peak levels of fecal coliforms in Mississippi Sound. Models consisting of one or more of the variables: Pearl River stage, water temperature, and salinity were developed to predict FC concentrations in the Sound. The model parameter(s) explained 56 to 91% of the variations in FC counts. Management of shellfish in Mississippi Sound can be improved by utilizing information on the forecasted three to seven years occurrence of ENSO events. In addition, since Pearl River stage was the most important variable predicting FC concentration in the Sound, a study of the levels and sources of FC bacteria in the river, especially the middle and lower sections, is needed for developing a management plan for reducing FC bacteria pollution in the Sound.
Chigbu, Paulinus; Gordon, Scott; Tchounwou, Paul B.
1. End-plate potentials (e.p.p.) have been recorded from the neuromuscular junctions of frog sartorius and extensor longus dig. IV muscles, using intracellular micropipettes. Either curare or MgCl2 were present in the Ringer solution, to keep the e.p.p. amplitude below the threshold for a muscle action potential and contraction. 2. It has been shown that saxitoxin (paralytic shellfish poison) usually caused a progressive reduction in the amplitude of the e.p.p. Occasionally, when it was applied in the presence of MgCl2, the e.p.p. disappeared abruptly. 3. Tetrodotoxin usually caused the e.p.p. to disappear abruptly. Occasionally, when applied in the presence of curare, the e.p.p. declined progressively for a short time before disappearing abruptly. 4. It is concluded that at the frog neuromuscular junction the preferential site of action of saxitoxin is at the nerve terminals, but tetrodotoxin preferentially blocks nerve conduction at a site proximal to the junction. 5. It is suggested that this preparation would be a convenient and reliable test object for distinguishing saxitoxin from tetrodotoxin.
Evans, M. H.
Phylogenetic and phenotypic analysis of cultivable marine bacteria isolated from laboratory cultures of two paralytic shellfish toxin-producing dinoflagellates, Gymnodinium catenatum and Alexandrium tamarense, showed the presence of a novel group of Gram-negative, aerobic, moderately halophilic and hydrocarbon-degrading bacteria, related to the genus Marinobacter. The strains, designated DG893T, DG1136 and ATAM407-13, grew optimally in media with 3-6 % NaCl and at 25-30 degrees C, and all could utilize n-hexadecane and n-tetradecane as the sole carbon source. The strains had a 16S rRNA gene sequence similarity of 94.2-94.3 % to Marinobacter hydrocarbonoclasticus ATCC 27132, and a similarity of 97.5-97.8 % to the closest phylogenetically related type strain, Marinobacter flavimaris DSM 16070T. DNA-DNA hybridization levels to M. flavimaris and other Marinobacter type strains were < or = 42 %, while DNA-DNA reassociation values among DG893T, DG1136 and ATAM407-13 were > or = 83 %. The DNA G + C content was 54-55 mol% and the major isoprenoid quinone was ubiquinone-9. On the basis of phenotypic, chemotaxonomic, DNA-DNA hybridization and phylogenetic analysis, it is proposed that these three strains represent a novel species, Marinobacter algicola sp. nov. The type strain is DG893T (= DSM 16394T = NCIMB 14009T). PMID:16514021
Green, David H; Bowman, John P; Smith, Elizabeth A; Gutierrez, Tony; Bolch, Christopher J S
Quantitative tools based on mechanistic modelling of functional traits able to enhance the sustainability of aquaculture and most other human activities (i.e. reducing the likelihood of detrimental impacts optimising productions), are especially important factors in the decision to site aquaculture facilities in coastal lakes, ponds and lagoons and, in the case of detrimental impact, to adopt mitigation measures. We tested the ability of mechanistic functional trait based models to predict life history traits of cultivable shellfish in shallow coastal lakes. Dynamic Energy Budget (DEB) models were run to generate spatially explicit predictions of Mytilus galloprovincialis life history (LH) traits (e.g. body size and fecundity). Using fortnightly data of food supply and hourly data of body temperatures, and exploiting the power of mechanistic rules, we estimated the amount of faeces ejected by a fixed quantity of organisms cultivated in two shallow Southern Mediterranean (Sicily) lakes. These differed in terms of temperature and food density, implying large differences in life history traits of mussels in the two study areas. This information could help facilitate the selection of sites where environmental conditions are more suitable for aquaculture and contextually compatible with sustainability. The validation exercise obtained by comparing the predicted and observed data was nearly consistent. Therefore, a mechanistic functional traits-based model seems able to capture the link between habitat characteristics and functional traits of organisms, delineating the fundamental portion of an ecological niche, the possibility of predicting LH traits and potential ecological applications in the management of natural coastal resources.
Rinaldi, A.; Montalto, V.; Manganaro, A.; Mazzola, A.; Mirto, S.; Sanfilippo, M.; Sarà, G.
The present study evaluate concentrations of arsenic (As), cadmium (Cd), chromium (Cr), lead (Pb), manganese (Mn), nickel (Ni), vanadium (V) and zinc (Zn) in fish and shellfish from the Gulf of Catania. Heavy metal analysis was carried on with an ICP-MS, and consumption rates advisory for minimizing chronic systemic and non cancer endpoints in child and adults have been estimated. Among metals investigated, only Cd and Pb have a limit set by European Community for human consumption, and the thresholds were not been exceeded in analyzed species. The As, toxic in its inorganic form, have not a regulatory limit yet, but the Environmental Protection Agency and the World Health Organization provide a reference dose, a cancer slope factor and a tolerable intake, applicable in the risk factors assessment. Arsenic target hazard quotient (THQ) values, suggest that human should minimizing meals/week of analyzed species to avoid deleterious effect during lifetime, furthermore, with As cancer risk assessment, for most of the fish, the risk for cancer is greater than the acceptable lifetime risk of 10(-5). Our results give important finding about the consumption limits on certain metals, especially for As, all for minimizing potential health risks in population. PMID:23211443
Copat, Chiara; Arena, Giovani; Fiore, Maria; Ledda, Caterina; Fallico, Roberto; Sciacca, Salvatore; Ferrante, Margherita
In 2002 several hundred people were taken ill after eating self-harvested brown crabs (Cancer pagurus) in the southern part of Norway. The symptoms were similar to diarrhetic shellfish poisoning (DSP) although with a somewhat delayed onset. This happened at the same time as an unusual early bloom of Dinophysis acuta had lead to high amounts of DSP toxins in blue mussels (Mytilus edulis) in the same area. The proposed cause of the intoxication was that crabs had accumulated toxins by eating blue mussels. Analyses of crab material from the area revealed very little free toxin in the form of okadaic acid (OA). However, after alkaline hydrolysis of the material, the amounts of OA found in the crabs were above the toxic level. MS/MS analysis of a sample from one intoxication episode indicated presence of the 14:0, 16:1, 16:0 and 18:1 fatty acid esters of okadaic acid. Esterified OA constituted more than 90% of total identified DSP toxins in crabs, indicating that not only esterified toxin from mussels was accumulated, but also that appreciable transfer of OA to OA-esters occurred in the crabs. PMID:16153454
Torgersen, Trine; Aasen, John; Aune, Tore
Sea otters Enhydra lutris can be bycaught and drowned in fishing pots and traps, which may pose a threat to the welfare of otter populations. We explored this potential problem and its solutions using a wide variety of analyses. We exposed live California (USA) sea otters to finfish traps, lobster traps, and mock Dungeness crab traps in captive trials and found that the animals attempted to enter the circular and rectangular fyke openings, with some becoming entrapped. Using both live and dead sea otters, we found that a 3 ?? 9 inch (7.6 ?? 22.9 cm) fyke opening (1 inch narrower than the 4 ?? 9 inch [10.2 ?? 22.9 cm] openings currently used in California's commercial Dungeness crab fishery) would exclude most free-living (i.e. weaned from their mothers) otters while permitting the undiminished capture of crabs. Observer programs do not currently exist in California for these fisheries, so we calculated the effort required by an observer program to document sea otter bycatch over a range of hypothetical levels and evaluated the impact of those mortality rates on population growth. These analyses demonstrate that significant mortality from bycatch might easily go undetected, even with seemingly high levels of observer effort. As sea otters reoccupy portions of their former habitat in California, co-occurrence with finfish and shellfish traps with relatively large fyke openings will increase. ?? Inter-Research 2011.
Hatfield, B. B.; Ames, J. A.; Estes, J. A.; Tinker, M. T.; Johnson, A. B.; Staedler, M. M.; Harris, M. D.
To investigate the potential for differential accumulation of paralytic shellfish toxins (PSTs) in various tissues of the akoya pearl oyster, Pinctada imbricata, two feeding trials were carried out using the PST-producing dinoflagellate, Alexandrium minutum. When fed with A. minutum at concentrations between 100 and 1300 cells ml(-1), the maximum clearance by P. imbricata was shown to occur at a density of 300 cells ml(-1). When fed twice daily at this rate for up 12 days, P. imbricata accumulated analogues of gonyautoxins (GTXs): GTXs 1,4 and 2,3. The levels of GTXs in the viscera increased progressively on days 4, 8 and 12 to peak at 17.9+/-4.47 microg STX-equivalent 100 g(-1) biomass. Following 12 days of depuration, in the absence of A. minutum, GTX levels fell by approximately 65% to 6.0+/-2.20 microg STX-equivalent 100 g(-1) biomass. No GTX was found in the oysters at the start of the trial or in untreated controls. The accumulation of GTX was found to be tissue specific. No GTX was detected in the muscle tissue of P. imbricata during the feeding trial. PMID:19375444
Murray, Shauna A; O'Connor, Wayne A; Alvin, Alfonsus; Mihali, Troco K; Kalaitzis, John; Neilan, Brett A
The main aim of this study was to determine the occurrence of resistant bacteria in florfenicol-treated and untreated scallop larval cultures from a commercial hatchery and to characterize some selected florfenicol-resistant strains. Larval cultures from untreated and treated rearing tanks exhibited percentages of copiotrophic bacteria resistant to florfenicol ranging from 0.03% to 10.67% and 0.49-18.34%, respectively, whereas florfenicol resistance among oligotrophic bacteria varied from 1.44% to 35.50% and 3.62-95.71%, from untreated and treated larvae, respectively. Florfenicol resistant microbiota from reared scallop larvae mainly belonged to the Pseudomonas and Pseudoalteromonas genus and were mainly resistant to florfenicol, chloramphenicol, streptomycin and co-trimoxazole. This is the first study reporting antimicrobial resistant bacteria associated to a shellfish hatchery and the results suggest that a continuous surveillance of antimicrobial resistance even in absence of antibacterial therapy is urgently required to evaluate potential undesirable consequences on the surrounding environments. PMID:23880028
Miranda, Claudio D; Rojas, Rodrigo; Garrido, Marcela; Geisse, Julieta; González, Gerardo
Neurotoxic shellfish poisoning and brevetoxin metabolites: a case study from Florida 1 1 The views, opinions, and\\/or findings contained herein are those of the authors and should not be construed as an official Department of the Army position, policy or decision unless so designated by other documentation
In June of 1996, three family members were diagnosed as suffering from neurotoxic shellfish poisoning (NSP) as a result of eating shellfish harvested from Sarasota Bay, Florida. Urine from two of these patients and extracts of shellfish collected from the same location were analyzed by radioimmunoassay (RIA) and by receptor binding assay. Activity consistent with brevetoxins was present in both
Mark A Poli; Steven M Musser; Robert W Dickey; Paul P Eilers; Sherwood Hall
This study was conducted to analyze the genetic variability of Escherichia coli from domesticated animal wastes for microbial source tracking (MST) application in fecal contaminated shellfish growing waters of Xiangshan Bay, East China Sea. (GTG)(5) primer was used to generate 1363 fingerprints from E. coli isolated from feces of known 9 domesticated animal sources around this shellfish culture area. Jackknife analysis of the complete (GTG)(5)-PCR DNA fingerprint library indicated that isolates were assigned to the correct source groups with an 84.28% average rate of correct classification. Based on one-year source tracking data, the dominant sources of E. coli were swine, chickens, ducks and cows in this water area. Moreover, annual and spatial changes of E. coli concentrations and host sources may affect the level and distribution of zoonotic pathogen species in waters. Our findings will further contribute to preventing fecal pollution in aquatic environments and quality control of shellfish. PMID:21645948
Fu, Ling-Lin; Shuai, Jiang-Bing; Wang, Yanbo; Ma, Hong-Jia; Li, Jian-Rong
Development of forecasting systems for harmful algal blooms (HABs) has been a long-standing research and management goal. Significant progress has been made in the Gulf of Maine, where seasonal bloom forecasts are now being issued annually using Alexandrium fundyense cyst abundance maps and a population dynamics model developed for that organism. Thus far, these forecasts have used terms such as “significant”, “moderately large” or “moderate” to convey the extent of forecasted paralytic shellfish poisoning (PSP) outbreaks. In this study, historical shellfish harvesting closure data along the coast of the Gulf of Maine were used to derive a series of bloom severity levels that are analogous to those used to define major storms like hurricanes or tornados. Thirty-four years of PSP-related shellfish closure data for Maine, Massachusetts and New Hampshire were collected and mapped to depict the extent of coastline closure in each year. Due to fractal considerations, different methods were explored for measuring length of coastline closed. Ultimately, a simple procedure was developed using arbitrary straight-line segments to represent specific sections of the coastline. This method was consistently applied to each year’s PSP toxicity closure map to calculate the total length of coastline closed. Maps were then clustered together statistically to yield distinct groups of years with similar characteristics. A series of categories or levels was defined (“Level 1: Limited”, “Level 2: Moderate”, and “Level 3: Extensive”) each with an associated range of expected coastline closed, which can now be used instead of vague descriptors in future forecasts. This will provide scientifically consistent and simply defined information to the public as well as resource managers who make decisions on the basis of the forecasts.
Kleindinst, Judith L.; Anderson, Donald M.; McGillicuddy, Dennis J.; Stumpf, Richard P.; Fisher, Kathleen M.; Couture, Darcie A.; Hickey, J. Michael; Nash, Christopher
Marine dinoflagellates of the genera Alexandrium are well known producers of the potent neurotoxic paralytic shellfish toxins that can enter the food web and ultimately present a serious risk to public health in addition to causing huge economic losses. Direct coastal monitoring of Alexandrium spp. can provide early warning of potential shellfish contamination and risks to consumers and so a rapid, sensitive, portable and easy-to-use assay has been developed for this purpose using an innovative planar waveguide device. The disposable planar waveguide is comprised of a transparent substrate onto which an array of toxin-protein conjugates is deposited, assembled in a cartridge allowing the introduction of sample, and detection reagents. The competitive assay format uses a high affinity antibody to paralytic shellfish toxins with a detection signal generated via a fluorescently labelled secondary antibody. The waveguide cartridge is analysed by a simple reader device and results are displayed on a laptop computer. Assay speed has been optimised to enable measurement within 15 min. A rapid, portable sample preparation technique was developed for Alexandrium spp. in seawater to ensure analysis was completed within a short period of time. The assay was validated and the LOD and CC? were determined as 12 pg/mL and 20 pg/mL respectively with an intra-assay CV of 11.3% at the CC? and an average recovery of 106%. The highly innovative assay was proven to accurately detect toxin presence in algae sampled from the US and European waters at an unprecedented cell density of 10 cells/L. PMID:23102433
Meneely, Julie P; Campbell, Katrina; Greef, Charles; Lochhead, Michael J; Elliott, Christopher T
Development of forecasting systems for harmful algal blooms (HABs) has been a long-standing research and management goal. Significant progress has been made in the Gulf of Maine, where seasonal bloom forecasts are now being issued annually using Alexandrium fundyense cyst abundance maps and a population dynamics model developed for that organism. Thus far, these forecasts have used terms such as “significant”, “moderately large” or “moderate” to convey the extent of forecasted paralytic shellfish poisoning (PSP) outbreaks. In this study, historical shellfish harvesting closure data along the coast of the Gulf of Maine were used to derive a series of bloom severity levels that are analogous to those used to define major storms like hurricanes or tornados. Thirty-four years of PSP-related shellfish closure data for Maine, Massachusetts and New Hampshire were collected and mapped to depict the extent of coastline closure in each year. Due to fractal considerations, different methods were explored for measuring length of coastline closed. Ultimately, a simple procedure was developed using arbitrary straight-line segments to represent specific sections of the coastline. This method was consistently applied to each year’s PSP toxicity closure map to calculate the total length of coastline closed. Maps were then clustered together statistically to yield distinct groups of years with similar characteristics. A series of categories or levels was defined (“Level 1: Limited”, “Level 2: Moderate”, and “Level 3: Extensive”) each with an associated range of expected coastline closed, which can now be used instead of vague descriptors in future forecasts. This will provide scientifically consistent and simply defined information to the public as well as resource managers who make decisions on the basis of the forecasts.
Kleindinst, Judith L.; Anderson, Donald M.; McGillicuddy, Dennis J.; Stumpf, Richard P.; Fisher, Kathleen M.; Couture, Darcie A.; Michael Hickey, J.; Nash, Christopher
Galicia's coast (northwestern Spain) is a major producer of bivalve molluscs. Over an 18-month period, the presence of Salmonella, Aeromonas, Plesiomonas shigelloides, Vibrio parahaemolyticus, and Clostridium botulinum was determined by PCR methods in mussels (22 batches) and infaunal bivalves (31 batches of clams and cockles) before and after depuration. All batches were harvested from Galician class B harvesting areas where bivalve molluscs must not exceed 4,600 Escherichia coli per 100 g of flesh and liquor in 90% of the samples. Virulence-associated genes of Salmonella (invA), Aeromonas (aerA, hlyA, alt, ast, and laf), P. shigelloides (hugA), V. parahaemolyticus (tdh and trh), and C. botulinum (BoNT) were not detected. The pR72H chromosomal DNA fragment, which is conservative in V. parahaemolyticus strains, was detected in five (4.7%) samples. A number of 192 suspect isolates did not fit the description of clinical Aeromonas phenospecies, pathogenic Vibrio spp., or P. shigelloides. The effectiveness of commercial depuration in reducing bacterial indicators was also examined. E. coli was reduced to < or = 230/100 g of flesh and liquor in 90.9% of mussel lots but in only 70.9% of infaunal bivalve lots. For total coliform elimination, mussels were also more effective. Total counts significantly (P < 0.005) correlated with numbers of Pseudomonas, Aeromonas, and Vibrio. Our data indicate that Salmonella and pathogenic bacteria indigenous to estuarine environments do not appear to be significant hazards in Galician molluscan shellfish. A reason for concern, however, is that clearance of E. coli to acceptable levels was not always achieved especially in infaunal bivalves. PMID:19681267
Martínez, O; Rodríguez-Calleja, J M; Santos, J A; Otero, A; García-López, M L
Maintaining food production while sustaining productive ecosystems is among the central challenges of our time, yet, it has been for millennia. Ancient clam gardens, intertidal rock-walled terraces constructed by humans during the late Holocene, are thought to have improved the growing conditions for clams. We tested this hypothesis by comparing the beach slope, intertidal height, and biomass and density of bivalves at replicate clam garden and non-walled clam beaches in British Columbia, Canada. We also quantified the variation in growth and survival rates of littleneck clams (Leukoma staminea) we experimentally transplanted across these two beach types. We found that clam gardens had significantly shallower slopes than non-walled beaches and greater densities of L. staminea and Saxidomus giganteus, particularly at smaller size classes. Overall, clam gardens contained 4 times as many butter clams and over twice as many littleneck clams relative to non-walled beaches. As predicted, this relationship varied as a function of intertidal height, whereby clam density and biomass tended to be greater in clam gardens compared to non-walled beaches at relatively higher intertidal heights. Transplanted juvenile L. staminea grew 1.7 times faster and smaller size classes were more likely to survive in clam gardens than non-walled beaches, specifically at the top and bottom of beaches. Consequently, we provide strong evidence that ancient clam gardens likely increased clam productivity by altering the slope of soft-sediment beaches, expanding optimal intertidal clam habitat, thereby enhancing growing conditions for clams. These results reveal how ancient shellfish aquaculture practices may have supported food security strategies in the past and provide insight into tools for the conservation, management, and governance of intertidal seascapes today.
Groesbeck, Amy S.; Rowell, Kirsten; Lepofsky, Dana; Salomon, Anne K.
Vibrio parahaemolyticus is a marine microorganism, recognized as an important cause of foodborne illness particularly in Asia, South America and United States. Outbreaks are rarely reported in Europe, but they can occur unexpectedly in relation, among other reasons, to the spread of highly virulent strains. It is known that the risk is proportional to exposure levels to pathogenic V. parahaemolyticus (i.e. carrying the tdh and/or the trh genes) but currently there is a lack of occurrence data for pathogenic V. parahaemolyticus in shellfish production areas of the Member States. In this study a total of 147 samples of bivalve molluscs, from harvesting areas of two Italian regions (Sardinia and Veneto) were analyzed for Escherichia coli and salmonella, according to Reg 2073/2005, and for detection and enumeration of total and toxigenic V. parahaemolyticus strains using a new DNA colony hybridization method. Environmental parameters (water temperature and salinity) were also recorded. Results of E. coli were consistently in agreement with the legislation limits for the harvesting class of origin and Salmonella was detected only in one sample. The average contamination levels for total V. parahaemolyticus were 84 and 73CFU/g respectively for Sardinia and Veneto, with the highest value reaching 8.7×10(3)CFU/g. Nineteen samples (12.9%) resulted positive for the presence of potentially pathogenic V. parahaemolyticus strains, with levels ranging between 10 and 120CFU/g and most of the positive samples (n=17) showing values equal or below 20CFU/g. A significant correlation (r=0.41) was found between water temperature and V. parahaemolyticus levels, as well as with isolation frequency. The data provided in this study on contamination levels of total and potentially pathogenic V. parahaemolyticus, seasonal distribution and correlation with water temperature, will help in defining appropriate monitoring programs and post-harvest policies for this hazard, improving the management of the harvesting areas and the safety of bivalve molluscs. PMID:24810197
Suffredini, Elisabetta; Mioni, Renzo; Mazzette, Rina; Bordin, Paola; Serratore, Patrizia; Fois, Federica; Piano, Annamaria; Cozzi, Loredana; Croci, Luciana
Maintaining food production while sustaining productive ecosystems is among the central challenges of our time, yet, it has been for millennia. Ancient clam gardens, intertidal rock-walled terraces constructed by humans during the late Holocene, are thought to have improved the growing conditions for clams. We tested this hypothesis by comparing the beach slope, intertidal height, and biomass and density of bivalves at replicate clam garden and non-walled clam beaches in British Columbia, Canada. We also quantified the variation in growth and survival rates of littleneck clams (Leukoma staminea) we experimentally transplanted across these two beach types. We found that clam gardens had significantly shallower slopes than non-walled beaches and greater densities of L. staminea and Saxidomus giganteus, particularly at smaller size classes. Overall, clam gardens contained 4 times as many butter clams and over twice as many littleneck clams relative to non-walled beaches. As predicted, this relationship varied as a function of intertidal height, whereby clam density and biomass tended to be greater in clam gardens compared to non-walled beaches at relatively higher intertidal heights. Transplanted juvenile L. staminea grew 1.7 times faster and smaller size classes were more likely to survive in clam gardens than non-walled beaches, specifically at the top and bottom of beaches. Consequently, we provide strong evidence that ancient clam gardens likely increased clam productivity by altering the slope of soft-sediment beaches, expanding optimal intertidal clam habitat, thereby enhancing growing conditions for clams. These results reveal how ancient shellfish aquaculture practices may have supported food security strategies in the past and provide insight into tools for the conservation, management, and governance of intertidal seascapes today. PMID:24618748
Groesbeck, Amy S; Rowell, Kirsten; Lepofsky, Dana; Salomon, Anne K
Shellfish carrying capacity is determined by the interaction of a cultured species with its ecosystem, which is strongly influenced by hydrodynamics. Water circulation controls the exchange of matter between farms and the adjacent areas, which in turn establishes the nutrient supply that supports phytoplankton populations. The complexity of water circulation makes necessary the use of hydrodynamic models with detailed spatial resolution in carrying capacity estimations. This detailed spatial resolution also allows for the study of processes that depend on specific spatial arrangements, e.g., the most suitable location to place farms, which is crucial for marine spatial planning, and consequently for decision support systems. In the present study, a fully spatial physical-biogeochemical model has been combined with scenario building and optimization techniques as a proof of concept of the use of ecosystem modeling as an objective tool to inform marine spatial planning. The object of this exercise was to generate objective knowledge based on an ecosystem approach to establish new mussel aquaculture areas in a Norwegian fjord. Scenario building was used to determine the best location of a pump that can be used to bring nutrient-rich deep waters to the euphotic layer, increasing primary production, and consequently, carrying capacity for mussel cultivation. In addition, an optimization tool, parameter estimation (PEST), was applied to the optimal location and mussel standing stock biomass that maximize production, according to a preestablished carrying capacity criterion. Optimization tools allow us to make rational and transparent decisions to solve a well-defined question, decisions that are essential for policy makers. The outcomes of combining ecosystem models with scenario building and optimization facilitate planning based on an ecosystem approach, highlighting the capabilities of ecosystem modeling as a tool for marine spatial planning. PMID:24988780
Filgueira, Ramon; Grant, Jon; Strand, Øivind
Paralytic Shellfish Toxins (PSTs) are highly toxic metabolic by-products of cyanobacteria and dinoflagellates. The filamentous cyanobacterium Lyngbya wollei produces a unique set of PSTs, including L. wollei toxins (LWT) 1-6. The accurate identification and quantification of PSTs from Lyngbya filaments is challenging, but critical for understanding toxin production and associated risk, as well as for providing baseline information regarding the potential for trophic transfer. This study evaluated several approaches for the extraction and analysis of PSTs from field-collected L. wollei dominated algal mats. Extraction of PSTs from lyophilized Lyngbya biomass was assessed utilizing hydrochloric acid and acetic acid at concentrations of 0.001-0.1 M. Toxin profiles were then compared utilizing two analysis techniques: pre-column oxidation (peroxide and periodate) High Performance Liquid Chromatography (HPLC) with Fluorescence (FL) detection and LC coupled with Mass Spectrometry (MS). While both acid approaches efficiently extracted PSTs, hydrochloric acid was found to convert the less toxic LWT into the more toxic decarbamoylgonyautoxins 2&3 (dcGTX2&3) and decarbamoylsaxitoxin (dcSTX). In comparison, extraction with 0.1 M acetic acid preserved the original toxin profile and limited the presence of interfering co-extractants. Although pre-chromatographic oxidation with HPLC/FL was relatively easy to setup and utilize, the method did not resolve the individual constituents of the L. wollei derived PST profile. The LC/MS method allowed characterization of the PSTs derived from L. wollei, but without commercially available LWT 1-6 standards, quantitation was not possible for the LWT. In future work, evaluation of the risk associated with L. wollei derived PSTs will require commercially available standards of LWT 1-6 for accurate determinations of total PST content and potency. PMID:22960450
Foss, Amanda J; Phlips, Edward J; Aubel, Mark T; Szabo, Nancy J
Mussels (Mytilus galloprovincialis) were fed cultures of the Paralytic Shellfish Poisoning agent Alexandrium minutum (Strain AL1V) for a 15-day period and, for the next 12 days, they were fed the non-toxic species Tetraselmis suecica, in order to monitor the intoxication/detoxification process. The toxin content in the bivalve was checked daily throughout the experiment. During the time-course of the experiment, the toxin profile of the bivalves changed substantially, showing increasingly greater differences from the proportions found in the toxigenic dinoflagellate used as food. The main processes involved in the accumulation of toxins and in the variation of the toxic profiles were implemented in a series of numerical models and the usefulness of those models to describe the actual intoxication/detoxification kinetics was assessed. Models that did not include transformations between toxins were unable to describe the kinetics, even when different detoxification rates were allowed for the toxins involved. The models including epimerization and reduction provided a good description of the kinetics whether or not differential detoxification was allowed for the different toxins, suggesting that the differences in detoxification rates between the toxins are not an important factor in regulating the change of the toxic profile. The implementation of Michaelis-Menten kinetics to describe the two reductive transformations produced a model that had a poorer fit to the data observed than the model that included only a first order kinetics. This suggests that, it is very unlikely that any enzymatic reaction is involved in the reduction of the hydroxycarbamate (OH-GTXs) to carbamate (H-GTXs) gonyautoxins. PMID:14757209
Blanco, Juan; Reyero, Ma Isabel; Franco, José
The aim of this work was to shed light on the anatomical distribution of diarrhetic shellfish poisoning (DSP) toxins in the mussel Mytilus galloprovincialis and to determine any possible changes undergone during the depuration process. To this end, the distribution of two DSP toxins--okadaic acid and DTX2--and some of their derivatives were studied by means of HPLC/MS at different stages of the depuration process. Mussels were collected from mussel farms located in the Galician Rías and they were collected under three types of circumstances: (a) while ingesting toxic phytoplankton cells; (b) 1 week after the toxic cells had disappeared from the water; and (c) after ca. 2 months of depuration. Additionally, in case (b), the distribution among tissues was checked every week over a depuration period of 35 days in the laboratory. DSP toxins were only detected in non-visceral tissues when the extracts were concentrated 20-fold and, even in these cases, the concentrations found were very low. When the maximum possible contribution of non-visceral tissues was computed, taking into account the technique's detection limits and tissue weight, no relevant contribution to the toxin burden of non-visceral tissues was found at any stage of depuration, with the maximum possible contributions usually below 7%. The concentrated samples analysed showed that the actual contribution in all the cases studied was, in fact, less than 1% of the total toxin burden. These findings suggest that (1) when analytical methods are used to monitor DSP toxic mussels, non-visceral tissues should be assumed to be free of toxins in order to precisely compute the toxin concentration of the whole mass of edible tissues and (2) when studying the accumulation kinetics of DSP toxins, transference from the digestive gland to other tissues should not be taken into account, as the other tissues do not contain relevant amounts of DSP toxins. PMID:17981314
Blanco, Juan; Mariño, Carmen; Martín, Helena; Acosta, Carmen P
A new analogue of okadaic acid (OA), the toxin mainly responsible for diarrhetic shellfish-poisoning (DSP) phenomena in Europe, has been isolated from toxic phytoplankton (Dinophysis acuta) collected in Irish waters. Fluorimetric LC analyses of the extracts of bulk phytoplankton samples using derivatisation with 9-anthryldiazomethane (ADAM) showed a complex toxin profile, with peaks corresponding to OA and dinophysistoxin-2 (DTX-2) as well as a third unidentified compound. This minor unidentified component was isolated by chromatographic techniques such as normal-phase chromatography, gel permeation on Sephadex, solid-phase extraction and reversed-phase separations. Ionspray mass spectrometry (MS) was used for structural investigation on this compound due to the very small amount of isolated material. Flow injection analysis (FIA)-MS of the isolated compound gave positive-ion mass spectrum dominated by the protonated molecule, [M + H]+, at signal m/z 805, whereas the deprotonated molecule [M - H]- was observed in the negative-ion spectrum at signal m/z 803, thus indicating the molecular weight of 804 for the new toxin, the same as OA and its known isomers, DTX-2 and DTX-2B. Collision-induced dissociation (CID) as obtained by positive and negative tandem mass spectrometry (MS-MS) showed a fragmentation pattern for the new compound which was very similar to that of OA, DTX-2 and DTX-2B. Ionspray microLC-MS of a mixture containing the compound under investigation together with OA analogues showed the compound eluted after OA, DTX-2, DTX-2B and before DTX-1. All the chromatographic and mass spectrometric data indicated the compound to be another OA isomer and it was therefore coded DTX-2C. To the best of our knowledge this is the first report on the isolation of a new compound related to DSP toxins from natural communities of toxic phytoplankton. PMID:9542135
Draisci, R; Giannetti, L; Lucentini, L; Marchiafava, C; James, K J; Bishop, A G; Healy, B M; Kelly, S S
In an attempt to feed purple clams (Hiatula rostrata) with dinoglagellate Alexandrium minutum, the maximal accumulation toxicity of paralytic shellfish poisoning (PSP) toxins reached 40.6 MU/g on day 5 of feeding. Subsequently, the toxicity increased no further, although purple clams ingested more toxic algae. Furthermore, when milkfish (Chanos chanos) larvae were treated with toxic, nontoxic A. minutum or PSP toxin-containing extract in the water medium, it was found that the mortality of fish increased with the increasing concentrations of toxic algae. PSP toxin-containing extract did not show any toxic effect on milkfish larvae. PMID:11695819
Chen, C Y
Okadaic acid (1) (OA) and its congeners are mainly responsible for diarrhetic shellfish poisoning (DSP) syndrome. The presence of several OA derivatives have already been confirmed in Prorocentrum and Dinophysis spp. In this paper, we report on the detection and identification of a new DSP toxin, the OA isomer 19-epi-okadaic acid (2) (19-epi-OA), isolated from cultures of Prorocentrum belizeanum, by determining its retention time (RT) and fragmentation pattern using liquid chromatography coupled with mass spectrometry (LC–MS/MS).
Paz, Beatriz; Daranas, Antonio H.; Cruz, Patricia G.; Franco, Jose M.; Norte, Manuel; Fernandez, Jose J.
A major goal in harmful algal bloom (HAB) research has been to identify mechanisms underlying interannual variability in bloom magnitude and impact. Here the focus is on variability in Alexandrium fundyense blooms and paralytic shellfish poisoning (PSP) toxicity in Maine, USA, over 34 years (1978-2011). The Maine coastline was divided into two regions - eastern and western Maine, and within those two regions, three measures of PSP toxicity (the percent of stations showing detectable toxicity over the year, the cumulative amount of toxicity per station measured in all shellfish (mussel) samples during that year, and the duration of measurable toxicity) were examined for each year in the time series. These metrics were combined into a simple HAB Index that provides a single measure of annual toxin severity across each region. The three toxin metrics, as well as the HAB Index that integrates them, reveal significant variability in overall toxicity between individual years as well as long-term, decadal patterns or regimes. Based on different conceptual models of the system, we considered three trend formulations to characterize the long-term patterns in the Index - a three-phase (mean-shift) model, a linear two-phase model, and a pulse-decline model. The first represents a “regime shift” or multiple equilibria formulation as might occur with alternating periods of sustained high and low cyst abundance or favorable and unfavorable growth conditions, the second depicts a scenario of more gradual transitions in cyst abundance or growth conditions of vegetative cells, and the third characterizes a ”sawtooth” pattern in which upward shifts in toxicity are associated with major cyst recruitment events, followed by a gradual but continuous decline until the next pulse. The fitted models were compared using both residual sum of squares and Akaike's Information Criterion. There were some differences between model fits, but none consistently gave a better fit than the others. This statistical underpinning can guide efforts to identify physical and/or biological mechanisms underlying the patterns revealed by the HAB Index. Although A. fundyense cyst survey data (limited to 9 years) do not span the entire interval of the shellfish toxicity records, this analysis leads us to hypothesize that major changes in the abundance of A. fundyense cysts may be a primary factor contributing to the decadal trends in shellfish toxicity in this region. The HAB Index approach taken here is simple but represents a novel and potentially useful tool for resource managers in many areas of the world subject to toxic HABs.
Anderson, Donald M.; Couture, Darcie A.; Kleindinst, Judith L.; Keafer, Bruce A.; McGillicuddy, Dennis J., Jr.; Martin, Jennifer L.; Richlen, Mindy L.; Hickey, J. Michael; Solow, Andrew R.
Lead, nickel, vanadium and cadmium were determined in fish and shellfish muscles, to assess contamination levels and identify bio-indicators. Vanadium was not detectable. Lead and cadmium were slightly above legal limits used in South East Asia, but lower than those of Australia and New Zealand. Higher contents of nickel, cadmium and lead in Pachymelania aurita and Crassostrea rhizophorae, lead in Mugil cephalus and cadmium in Periophthalmus koelreuteri, mark these species out as possible bio-indicators for the three metals in aquatic systems. Patterns of bioaccumulation seem to suggest that biophysiological and ecological characteristics influence bioaccumulation of trace metals in fish and shell fish. PMID:23568331
Marcus, A C; Okoye, C O B; Ibeto, C N
Suitability of postcolumn oxidation liquid chromatography method AOAC 2011.02 for monitoring paralytic shellfish toxins in Alaskan shellfish--initial pilot study versus mouse bioassay and in-house validation.
An in-house study was conducted to confirm the suitability of the postcolumn oxidation (PCOX) LC method, AOAC 2011.02, for regulatory monitoring of paralytic shellfish toxins (PSTs) in Alaskan shellfish. The following parameters were evaluated: calibration linearity, linear range, retention time stability, accuracy, repeatability, LOQ, and comparability with AOAC 959.08, the mouse bioassay (MBA) for PSTs. Mean recoveries for oyster homogenate spiked at 66 microg saxitoxin (STX) diHCI eq./100 g and 5 microg STX diHCI eq./100 g were 95 (n = 8) and 104%, respectively. Repeatability (n = 8) was 1.6 and 7% RSD, respectively. LOQ was estimated at 5 microg STX diHCl eq./100 g with S/N = 6 for STX. Comparability with the MBA was evaluated by duplicate analysis of regulatory samples using the original homogenate for both methods. Forty-one samples with MBA results varying between 40 and 500 microg STX diHCl eq./100 g were compared to corresponding PCOX values. The correlation coefficient (r2) = 0.96, with a slope of 2.1. The slope indicates an average 2X higher total toxicity result for PCOX versus MBA, a result that is consistent with prior literature showing low recoveries using the MBA. PMID:24830139
During 2006 and 2007, we documented the re-emergence of severe episodes of vibriosis caused by Vibrio tubiashii in shellfish hatcheries on the west coast of North America. Lost larval and juvenile production included 3 previously undescribed hosts, Pacific (Crassostrea gigas) and Kumamoto (C. sikamea) oysters and geoduck clams Panope abrupta, with a 2007 decline in larval oyster production of approximately 59% in one hatchery. Losses of larval and juvenile bivalves were linked to V. tubiashii blooms in the coastal environment, which were associated with the apparent mixing of unusually warm surface seawater and intermittently upwelled cooler, nutrient- and Vibrio spp.- enriched seawater. The ocean temperature elevation anomaly in 2007 was not clearly linked to an El Niño event, as was a similar episode in 1998. Concentrations of the dominant shellfish-pathogenic vibrios were as high as 1.6 x 10(5) cfu ml(-1) in the cold, upwelled water. The bacteria possessed the genes coding for a protease and hemolysin described for V. tubiashii, and pathogenic isolates secreted these peptides. Lesions resulting from a classic invasive disease and a toxigenic noninvasive disease occurred in oyster and geoduck clam larvae. Management and prevention require reduction of incoming concentrations of the bacteria, reduction of contamination in water and air supplies and in stock chemical solutions, removal of bacterial toxins, and interruption of the cycle of bacterial amplification in the hatchery and in microalgal food supplies. PMID:19149375
Elston, Ralph A; Hasegawa, Hiroaki; Humphrey, Karen L; Polyak, Ildiko K; Häse, Claudia C
The combustion of fossil fuels has enriched levels of CO2 in the world’s oceans and decreased ocean pH. Although the continuation of these processes may alter the growth, survival, and diversity of marine organisms that synthesize CaCO3 shells, the effects of ocean acidification since the dawn of the industrial revolution are not clear. Here we present experiments that examined the effects of the ocean’s past, present, and future (21st and 22nd centuries) CO2 concentrations on the growth, survival, and condition of larvae of two species of commercially and ecologically valuable bivalve shellfish (Mercenaria mercenaria and Argopecten irradians). Larvae grown under near preindustrial CO2 concentrations (250 ppm) displayed significantly faster growth and metamorphosis as well as higher survival and lipid accumulation rates compared with individuals reared under modern day CO2 levels. Bivalves grown under near preindustrial CO2 levels displayed thicker, more robust shells than individuals grown at present CO2 concentrations, whereas bivalves exposed to CO2 levels expected later this century had shells that were malformed and eroded. These results suggest that the ocean acidification that has occurred during the past two centuries may be inhibiting the development and survival of larval shellfish and contributing to global declines of some bivalve populations.
Gobler, Christopher J.
Paralytic shellfish poisoning is a toxic syndrome described in humans following the ingestion of seafood contaminated with saxitoxin and/or its derivatives. The presence of these toxins in shellfish is considered an important health threat and their levels in seafood destined to human consumption are regulated in many countries, as well as the levels of other chemically unrelated toxins. We studied the feasibility of immunodetection of saxitoxin and its analogs using a solid-phase microsphere assay coupled to flow cytometry detection in a Luminex 200 system. The technique consists of a competition assay where the toxins in solution compete with bead-bound saxitoxin for binding to an antigonyautoxin 2/3 monoclonal antibody (GT-13A). The assay allowed the detection of saxitoxin both in buffer and mussel extracts in the range of 2.2-19.7 ng/mL (IC(20)-IC(80)). Moreover, the assay cross-reactivity with other toxins of the group is similar to previously published immunoassays, with adequate detection of most analogs except N-1 hydroxy analogs. The recovery rate of the assay for saxitoxin was close to 100%. This microsphere-based immunoassay is suitable to be used as a screening method, detecting saxitoxin from 260 to 2360 ?g/kg. This microsphere/flow cytometry system provided similar sensitivities to previously published immunoassays and provides a solid background for the development of easy, flexible multiplexing of toxin detection in one sample. PMID:22500610
Fraga, María; Vilariño, Natalia; Louzao, M Carmen; Campbell, Katrina; Elliott, Christopher T; Kawatsu, Kentaro; Vieytes, Mercedes R; Botana, Luis M
Three assays were developed to enumerate total and fecal Escherichia coli and total Vibrionaceae in shellfish, seawater, and other foods and environmental samples. Assays involve membrane overlays of overnight colonies on nonselective agar plates to detect beta-glucuronidase and lysyl aminopeptidase activities for E. coli and Vibrionaceae, respectively. Cellulose membranes containing the substrates 4-methylumbeferyl-beta-D-glucuronide (MUG) produced a bright blue fluorescence when overlaid onto E. coli, while L-lysyl-7-amino-4-trifluoromethylcoumarin produced green fluorescent foci when overlaid onto Vibrionaceae family members. A multiplex assay was also developed for simultaneously enumerating total E. coli and total Vibrionaceae in oysters and seawater. Overall, 65% of overlaid E. coli (non-O157:H7) were MUG-positive, compared with 62% as determined by the most-probable-number-MUG assay. The overlays are rapid, simple, and cost effective for quantification purposes. This research provides practical alternatives for monitoring bacterial indicators and potential pathogens in complex samples, including molluscan shellfish. PMID:20396663
Richards, Gary P; Watson, Michael A
Surfclams, Spisula solidissima, pose a particular health risk for human consumption as they are characterized by accumulation of extremely high levels of toxins associated with paralytic shellfish poisoning (PSP), slow toxin elimination and an extremely high post-ingestive capacity for toxin bioconversion. Surfclam populations experience a wide range of temperatures along the NW Atlantic continental shelf, and are undergoing range contraction that has been attributed to global warming. In this study the influence of temperature (5, 12 and 21 °C) on detoxification kinetics of individual PSP toxins in two tissue compartments of juvenile surfclams (?35 mm shell length) was determined under controlled laboratory conditions, over prolonged (2.4 months) depuration. Clams were toxified with a representative regional Gulf of Maine isolate of the dinoflagellate Alexandrium fundyense of known toxin profile, allowing tracking of changes in toxin composition and calculated toxicity in surfclam tissues. The visceral mass detoxified at all temperatures, although toxin loss rate increased with increasing temperature. In contrast, total toxin content and calculated toxicities in other tissues remained constant or even increased during depuration, suggesting a physiological or biochemical toxin-retention mechanism in this tissue pool and temperature-independent detoxification. In vivo toxin compositional changes in surfclam tissues found in this study provide evidence of specific toxin conversion pathways, involving both reductive and decarbamoylation pathways. We conclude that such toxin biotransformations, especially in non-visceral tissues, may introduce a discrepancy in describing kinetics of total toxicity (in saxitoxin equivalents [STXeq]) of S. solidissima over prolonged detoxification. Nevertheless, use of total toxicity values generated by routine regulatory monitoring based upon mouse bioassays or calculated from chemical analytical determination of molar toxin concentrations is adequate for first-order modeling of toxin kinetics in this species. Furthermore, the differential detoxification response of viscera and other tissues in relation to temperature emphasizes the need for two-compartment modeling to describe the fate of PSP toxins in this species. Finally, key parameters were identified that may prove useful in hindcasting the timing of toxic blooms or new toxin input in deep offshore waters where routine monitoring of toxic phytoplankton is impractical.
Monica Bricelj, V.; Cembella, Allan D.; Laby, David
Paralytic shellfish poisoning toxins (PSTs) are a family of more than 30 natural alkaloids synthesized by dinoflagellates and cyanobacteria whose toxicity in animals is mediated by voltage-gated Na(+) channel blocking. The export of PST analogues may be through SxtF and SxtM, two putative MATE (multidrug and toxic compound extrusion) family transporters encoded in PSTs biosynthetic gene cluster (sxt). sxtM is present in every sxt cluster analyzed; however, sxtF is only present in the Cylindrospermopsis-Raphidiopsis clade. These transporters are energetically coupled with an electrochemical gradient of proton (H(+)) or sodium (Na(+)) ions across membranes. Because the functional role of PSTs remains unknown and methods for genetic manipulation in PST-producing organisms have not yet been developed, protein structure analyses will allow us to understand their function. By analyzing the sxt cluster of eight PST-producing cyanobacteria, we found no correlation between the presence of sxtF or sxtM and a specific PSTs profile. Phylogenetic analyses of SxtF/M showed a high conservation of SxtF in the Cylindrospermopsis-Raphidiopsis clade, suggesting conserved substrate affinity. Two domains involved in Na(+) and drug recognition from NorM proteins (MATE family) of Vibrio parahaemolyticus and V. cholerae are present in SxtF/M. The Na(+) recognition domain was conserved in both SxtF/M, indicating that Na(+) can maintain the role as a cation anti-transporter. Consensus motifs for toxin binding differed between SxtF and SxtM implying differential substrate binding. Through protein modeling and docking analysis, we found that there is no marked affinity between the recognition domain and a specific PST analogue. This agrees with our previous results of PST export in R. brookii D9, where we observed that the response to Na(+) incubation was similar to different analogues. These results reassert the hypothesis regarding the involvement of Na(+) in toxin export, as well as the motifs L(398)XGLQD(403) (SxtM) and L(390)VGLRD(395) (SxtF) in toxin recognition. PMID:23457475
Soto-Liebe, Katia; López-Cortés, Xaviera A; Fuentes-Valdes, Juan José; Stucken, Karina; Gonzalez-Nilo, Fernando; Vásquez, Mónica
Four species of shellfish (Mytilus galloprovincialis, Ostrea edulis, Chlamis varia and Venus verrucosa) were collected during the autumn 2011 and spring 2012 along the eastern Adriatic coast from six shellfish harvesting areas (all species) and 13 breeding sampling areas (mussels) to assess As, Cd, Hg and Pb levels and the human risks of shellfish consumption. The mean metal concentrations (wet weight) in the examined species ranged from 1.420 to 9.575mg/kg for As, 0.034 to 1.270 for Cd, 0.005 to 0.680 for Hg and 0.140 to 2.072 for Pb. Examination of the spatial distribution of As, Cd, Hg and Pb revealed statistically significant differences among the studied areas. Since the concentrations were below the maximum prescribed by the laws of the EU and Croatia (the concentrations slightly exceeded the upper limits for three samples; Pb, Cd and Hg) and the hazard index, (HI) for Cd, Hg and Pb were below 1 and the target cancer risk (TR) for As was lower than 1×10(-6), there is no human health risk of consumption of shellfish from Croatian waters. PMID:24176332
Bogdanovi?, Tanja; Ujevi?, Ivana; Sedak, Marija; Listeš, Eddy; Simat, Vida; Petri?evi?, Sandra; Poljak, Vedran
Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT-PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples.
Jothikumar, Narayanan; Lowther, James A.; Henshilwood, Kathleen; Lees, David N.; Hill, Vincent R.; Vinje, Jan
The ability of two Alexandrium species to produce paralytic shellfish toxins (PST) in laboratory culture following the generation of bacteria-free cultures was investigated. The dinoflagellates Alexandrium lusitanicum NEPCC 253 and Alexandrium tamarense NEPCC 407 were cultured in the presence of antibiotics and tested for residual bacteria. After treatment with a cocktail of streptomycin, ciprofloxacin, gentamicin and penicillin G, bacteria could
Georgina L Hold; Elizabeth A Smith; T. Harry Birkbeck; Susan Gallacher
Paralytic shellfish poisoning (PSP), a potentially fatal foodborne illness, is often diagnosed anecdotally based on symptoms and dietary history. The neurotoxins responsible for PSP, collectively referred to as the saxitoxins or paralytic shellfish toxins (PSTs), are natural toxins, produced by certain dinoflagellates, that may accumulate in seafood, particularly filter-feeding bivalves. Illnesses are rare because of effective monitoring programs, yet occasional poisonings occur. Rarely are contaminated food and human clinical samples (e.g., urine and serum) available for testing. There are currently few methods, none of which are validated, for determining PSTs in clinical matrices. This study evaluated AOAC (Association of Analytical Communities) Official Method of Analysis (OMA) 2005.06. [AOAC Official Method 2005.06 Paralytic Shellfish Poisoning Toxins in Shellfish: Prechormatographic Oxidation and Liquid Chromatography with Fluorescence Detection. In Official Methods of Analysis of AOAC International
DeGrasse, Stacey; Rivera, Victor; Roach, John; White, Kevin; Callahan, John; Couture, Darcie; Simone, Karen; Peredy, Tamas; Poli, Mark
The rep-PCR DNA fingerprinting performed with REP, BOX A1R, and (GTG)(5) primers was investigated as a way to differentiate between human, livestock, and poultry sources of fecal pollution on the area of Xiangshan Bay, East China Sea. Of the three methods, the BOX-PCR DNA fingerprints analyzed by jack-knife algorithm were revealed high rate of correct classification (RCC) with 91.30, 80.39, 89.39, 86.14, 93.24, 87.72, and 89.28% of human, cattle, swine, chicken, duck, sheep, and goose E. coli isolates classified into the correct host source, respectively. The average rate of correct classification (ARCC) of REP-, BOX-, and (GTG)(5)-PCR patterns was 79.88, 88.21, and 86.39%, respectively. Although the highest amount of bands in (GTG)(5)-PCR fingerprints could be observed, the discriminatory efficacy of BOX-PCR was superior to both REP- and (GTG)(5)-PCR. Moreover, the similarity of 459 isolates originated from shellfish and growing water was compared with fecal-obtained strains. The results showed that 92.4 and 96.2% E. coli strains isolated from midstream and downstream shellfish samples, respectively, had a ? 80% similarity with corresponding strains isolated from fecal samples. It was indicated that E. coli in feces could spread from human sewage or domestic farms to the surrounding shellfish culture water, and potentially affect the quality of shellfish. This work suggests that rep-PCR fingerprinting can be a promising genotypic tool applied in the shellfish growing water management on East China Sea for source identification of fecal pollution. PMID:21279641
Ma, Hong-Jia; Fu, Ling-Lin; Li, Jian-Rong
This study was conducted to quantitatively determine the fatty acid contents of 20 species of marine fish and four species of shellfish from Straits of Malacca. Most samples contained fairly high amounts of polyunsaturated fatty acids (PUFAs), especially alpha-linolenic acid (ALA, C18:3 n3), eicosapentaenoic acid (EPA, C20:5 n3), and docosahexaenoic acid (DHA, C22:6 n3). Longtail shad, yellowstripe scad, and moonfish contained significantly higher (P < 0.05) amounts of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and alpha-linolenic acid (ALA), respectively. Meanwhile, fringescale sardinella, malabar red snapper, black pomfret, Japanese threadfin bream, giant seaperch, and sixbar grouper showed considerably high content (537.2-944.1?mg/100 g wet sample) of desirable omega-3 fatty acids. The polyunsaturated-fatty-acids/saturated-fatty-acids (P/S) ratios for most samples were higher than that of Menhaden oil (P/S = 0.58), a recommended PUFA supplement which may help to lower blood pressure. Yellowstripe scad (highest DHA, ? - 3/? - 6 = 6.4, P/S = 1.7), moonfish (highest ALA, ? - 3/? - 6 = 1.9, P/S = 1.0), and longtail shad (highest EPA, ? - 3/? - 6 = 0.8, P/S = 0.4) were the samples with an outstandingly desirable overall composition of fatty acids. Overall, the marine fish and shellfish from the area contained good composition of fatty acids which offer health benefits and may be used for nutraceutical purposes in the future. PMID:23509703
Abd Aziz, Nurnadia; Azlan, Azrina; Ismail, Amin; Mohd Alinafiah, Suryati; Razman, Muhammad Rizal
Glutathione-S-transferases have been identified in all the living species examined so far, yet little is known about their function in marine organisms. In a previous report, the recently identified GST from Antarctic bivalve Laternula elliptica (LeGST) was classified into the rho class GST, but there are several unique features of LeGST that may justify reclassification, which could represent specific shellfish GSTs. Here, we determined the crystal structure of LeGST, which is a shellfish specific class of GST. The structural analysis showed that the relatively open and wide hydrophobic H-site of the LeGST allows this GST to accommodate various substrates. These results suggest that the H-site of LeGST may be the result of adaptation to their environments as sedentary organisms. PMID:23152139
Park, Ae Kyung; Moon, Jin Ho; Jang, Eun Hyuk; Park, Hyun; Ahn, In Young; Lee, Ki Seog; Chi, Young Min
Contamination of shellfish with paralytic shellfish poisoning toxins (PST) produced by toxic harmful algal blooms (HABs) have been negatively affecting the shellfish and aquaculture industries worldwide. Therefore, accurate and early identification of toxic phytoplankton species is crucial in HABs surveillance programs that allow fish-farmers to take appropriate preventive measures in shellfish harvesting and other aquaculture activities to overcome the negative impacts of HABs on human health. The identification of toxic dinoflagellates present in the water is currently a time-consuming operation since it requires skillful taxonomists and toxicologists equipped with optical and scanning electron microscopes as well as sophisticated equipment, for example, high-performance liquid chromotography-fluorescence detection. In this paper, a two-dimensional gel electrophoresis (2-DE)-based proteomic approach was applied to discriminate between toxic and nontoxic strains of Alexandrium minutum. Variation in morphological features between toxic and nontoxic strains was minimal and not significant. Also, variation in 2-DE protein patterns within either toxic or nontoxic strains was low, but pronounced differences were detected between toxic and nontoxic strains. The most notable differences between these strains were several abundant proteins with pIs ranging from 4.8 to 5.3 and apparent molecular masses between 17.5 and 21.5 kDa. Groups of proteins, namely NT1, NT2, NT3, and NT4, were consistently found in all nontoxic strains, while T1 and T2 were prominent in the toxic strains. These specific protein spots characteristic for toxic and nontoxic strains remained clearly distinguishable irrespective of the various growth conditions tested. Therefore, they have the potential to serve as "taxonomic markers" to distinguish toxic and nontoxic strains within A. minutum. Initial studies revealed that the expression pattern of T1 was tightly correlated to toxin biosynthesis in the examined alga and may be used to serve as a potential toxin indicator. PMID:15800974
Chan, Leo Lai; Hodgkiss, Ivor John; Lam, Paul Kwan-Sing; Wan, Jennifer Man-Fan; Chou, Hong-Nong; Lum, John Hon-Kei; Lo, Maria Gar-Yee; Mak, Abby Sin-Chi; Sit, Wai-Hung; Lo, Samuel Chun-Lap
The contamination of different types of shellfish by okadaic acid (OA)-group toxin esters is an important problem that presents serious risk for human health. During previous investigations carried out in our laboratory by liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS), the occurrence of a high percentage of esters in relation to the total OA equivalents has been observed in several shellfish species. The determination of these kinds of toxins using LC/MS or other chemical methods requires a hydrolysis step in order to convert the sterified compounds into the parent toxins, OA, dinophysistoxins-1 (DTX-1) and dinophysistoxins-2 (DTX-2). Most of the hydrolysis procedures are based on an alkaline hydrolysis reaction. However, despite hydrolysis being a critical step within the analysis, it has not been studied in depth up to now. The present paper reports the results obtained after evaluating the hydrolysis process of an esterified form of OA by using a standard of 7-O-acyl ester with palmitoyl as the fatty acid (palOA). Investigations were focused on checking the effectiveness of the hydrolysis for palOA using methanol as solvent standard and matrices matched standards. From the results obtained, no matrix influence on the hydrolysis process was observed and the quantity of palOA converted into OA was always above 80%. The analyses of different Spanish shellfish samples showed percentages of palOA in relation to the total OA esters ranging from 27% to 90%, depending on the shellfish specie. PMID:18243269
Villar-González, A; Rodríguez-Velasco, M L; Ben-Gigirey, B; Yasumoto, T; Botana, L M
This bibliography contains citations concerning heavy-metal contamination of shellfish and marine plants. Toxicity levels and the long-term effects on the ecology of the marine environment are discussed. The growth rate of marine life as a function of metal concentration and the long-term effects on the food chain are also considered. (This updated bibliography contains 276 citations, 13 of which are new entries to the previous edition.)
Amnesic shellfish poisoning (ASP) is a potentially lethal human toxic syndrome which is caused by domoic acid (DA) that originates in marine phytoplankton belonging to the Pseudonitzschia genus. A confirmatory and sensitive procedure has been developed and validated for the determination of DA in shellfish. The proposed method includes pressurised liquid extraction (PLE) with methanol/acetone (9:1), florisil clean-up purification inside the PLE extraction cell and detection by liquid chromatography (LC) coupled to electrospray ionization in positive mode tandem mass spectrometry (ESI-MS-MS). Comparison of ionization sources (ESI, atmospheric pressure ionization (APCI) atmospheric pressure photoionization (APPI) and combined APCI/APPI) were carried out in order to improve the analytical signal. The main parameters affecting the performance of the different ionization sources and PLE parameters were previously optimised using statistical design of experiments (DOE). Linear calibrations were obtained using mussel tissue extracts 0.05-5 microg DA/ml (R2>0.999). The limits of detection (LOD) and quantitation (LOQ) of the method were 0.2 and 0.5 microg/g respectively and recoveries ranged from 81 to 95%. This method was successfully applied to determine DA levels in 46 shellfish samples collected from Valencian (Spain) supermarkets, showing high sample throughput. PMID:17475272
Pardo, Olga; Yusà, Vicent; León, Nuria; Pastor, Agustín
This work proposes a procedure for the determination of total selenium content in shellfish after digestion of samples in block using cold finger system and detection using atomic fluorescent spectrometry coupled hydride generation (HG AFS). The optimal conditions for HG such as effect and volume of prereduction KBr 10 % (m/v) (1.0 and 2.0 ml) and concentration of hydrochloric acid (3.0 and 6.0 mol L(-1)) were evaluated. The best results were obtained using 3 mL of HCl (6 mol L(-1)) and 1 mL of KBr 10 % (m/v), followed by 30 min of prereduction for the volume of 1 mL of the digested sample. The precision and accuracy were assessed by the analysis of the Certified Reference Material NIST 1566b. Under the optimized conditions, the detection and quantification limits were 6.06 and 21.21 ?g kg(-1), respectively. The developed method was applied to samples of shellfish (oysters, clams, and mussels) collected at Todos os Santos Bay, Bahia, Brazil. Selenium concentrations ranged from 0.23?±?0.02 to 3.70?±?0.27 mg kg(-1) for Mytella guyanensis and Anomalocardia brasiliana, respectively. The developed method proved to be accurate, precise, cheap, fast, and could be used for monitoring Se in shellfish samples. PMID:24771464
Lopes Dos Santos, Walter Nei; Macedo, Samuel Marques; Teixeira da Rocha, Sofia Negreiros; Souza de Jesus, Caio Niela; Cavalcante, Dannuza Dias; Hatje, Vanessa
This paper describes a gas chromatography-mass spectrometric method of l-hydroxypyrene (1-HOP) and 2-naphthol (2-NAP) in fish and shellfish. Alkali hydrolysis method in this study was chosen and optimized to the reaction condition for 90 min at 90°C in a 2.0M KOH solution. For five independent determinations at 0.2 and 1.0 ?g/kg, the coefficient of variation was less than 5.1%. This method was used to assess the long-term influence of spilled crude oil on marine ecosystems and analyze fifty-two shellfish samples taken in the near of the accident region of the Hebei Spirit oil spill. 2-NAP and 1-HOP were detected in the mean concentration range of 0.09-12.42 and 0.03-0.06 ?g/kg, respectively. 2-NAP was detected in a high concentration range in shellfishes gathered in 2 months after the accident and it decreased rapidly to 6 months after that. The results showed that 2-NAP might be an important biomarker in biota contaminated with crude oil. PMID:23411178
Lim, Hyun-Hee; Shin, Ho-Sang
This study describes an experience in the training of female shellfish gatherers in the fishing community in Ilha do Paty, São Francisco do Conde, within the context of actions to promote health and food safety. This is an intervention study with planning of activities by a multidisciplinary team. The activities were developed in five stages: survey of topics of interest; awareness; female shellfish gatherers' work routine; teaching of best practices in the processing of shellfish; and group conversation. The methodologies included exposure through dialogue, group dynamics, workshops, theater, work with images, directed activities and the distribution of educational materials. At the end, an evaluation by the participants regarding the different aspects was conducted. Overall there was satisfaction among the audience for most indicators, which may relate to both the use of audiovisual resources and strategies that permitted the exchange of experiences regarding the recognition of the importance of training by the target audience. The experiment demonstrates the potential of developing educational activities with the fishing communities, with results that contribute to the bolstering of the local supply chain, with the promotion of food safety and occupational health. PMID:24897221
Nóbrega, Gabriela Silva da; Cardoso, Ryzia de Cassia Vieira; Furtunato, Dalva Maria da Nóbrega; Góes, José Ângelo Wenceslau; Ferreira, Tereza Cristina Braga; Santos, Mary Daiane Fontes; Santos, Sissa Maria Garrido
As part of an effort to develop a broadly applicable test for Norwalk-like viruses and hepatitis A virus (HAV) in shellfish, a rapid extraction method that is suitable for use with one-step reverse transcription (RT)-PCR-based detection methods was developed. The method involves virus extraction using a pH 9.5 glycine buffer, polyethylene glycol (PEG) precipitation, Tri-reagent, and purification of viral poly(A) RNA by using magnetic poly(dT) beads. This glycine–PEG–Tri-reagent–poly(dT) method can be performed in less than 8 h on hard-shell clams (Mercenaria mercenaria) and Eastern oysters (Crassostrea virginica) and, when coupled with RT-PCR-based detection, can yield results within 24 h. Observed sensitivities for seeded shellfish extracts are as low as 0.015 PFU of HAV and 22.4 RT-PCR50 units for Norwalk virus. Detection of HAV in live oysters experimentally exposed to contaminated seawater is also demonstrated. An adaptation of this method was used to identify HAV in imported clams (tentatively identified as Ruditapes philippinarum) implicated in an outbreak of food-borne viral illness. All of the required reagents are commercially available. This method should facilitate the implementation of RT-PCR testing of commercial shellfish.
Kingsley, David H.; Richards, Gary P.
Natural abundance carbon isotope analyses are sensitive tracers for fates and use of oil in aquatic environments. Use of oil carbon in estuarine food webs should lead to isotope values approaching those of oil itself, -27‰ for stable carbon isotopes reflecting oil origins and -1000‰ for carbon-14 reflecting oil age. To test for transfer of oil from the 2010 Deepwater Horizon spill into estuarine food webs, filter-feeding barnacles (Balanus sp.) and marsh mussels (Geukensia demissa) were collected from Louisiana estuaries near the site of the oil spill. Carbon-14 analyses of these animals from open waters and oiled marshes showed that oil use was <1% and near detection limits estimated at 0.3% oil incorporation. Respiration studies showed no evidence for enhanced microbial activity in bay waters. Results are consistent with low dietary impacts of oil for filter feeders and little overall impact on respiration in the productive Louisiana estuarine systems. PMID:24461698
Fry, Brian; Anderson, Laurie C
The periodic occurrence of Paralytic Shellfish Poisoning (PSP) toxins in UK surf clams and the recent move away from biological assays for PSP testing resulted in the need to determine method performance characteristics for the replacement analytical method in this species. With the requirement for laboratory reference materials to aid this validation together with known issues relating to toxin transformation in live clams and homogenised tissue, there was the need to assess the toxin transformation characteristics of PSP toxins in surf clam tissue. Initial work examined the rates of toxin transformation in UK surf clam tissue incubated with toxin standards, showing rapid transformation of N-sulfocarbamoyl toxins with slower transformation of carbamate toxins. Full transformational pathways were determined using a combination of three different analytical methods and confirmed the major expected transformations involving decarbamoylation, with some evidence for additional reaction pathways. Results obtained from the analysis of surf clam and oyster tissues incubated with varying concentrations of toxic Alexandrium algae highlighted expected transformation reactions, although significant differences were observed in the extent of the transformations amongst the range of toxins studied, with less efficient transformation of N-hydroxylated toxins as compared with other carbamate and N-sulfocarbamoyl toxins. Analysis of PSP-toxic incurred oyster, scallop and mussel tissues incubated with variable proportions of surf clam tissue showed large differences in the extent of the transformations. Total conversion of N-sulfocarbamoyl toxins was confirmed at low relative proportions of surf clam tissue in all three species, whereas transformation of carbamate toxins was found to occur only in the presence of higher proportions of surf clam tissue in oysters and mussels in comparison with scallops. Results enabled the production of three laboratory reference materials prepared following incubation of incurred homogenates with optimum proportions of surf clam tissue, resulting in materials containing a large number of PSP toxins. Stability experiments provided good preliminary evidence for the stability of these targeted materials under storage conditions. The work therefore provides both additional information relating to the transformational activity in UK surf clams and highlights a good potential method for the targeted production of reference materials which include a wider range of toxins than normally present in naturally incurred shellfish. PMID:23369833
Turner, Andrew D; Lewis, Adam M; O'Neil, Alison; Hatfield, Robert G
Runoff of cattle manures (cows, pigs, sheeps) or discharge of effluent from wastewater treatment plants (WWTP) into aquatic ecosystems can lead to microbiological contamination of waters and living organisms. In coastal ecosystems and particularly in shellfish harvesting areas, the presence of pathogen microorganisms in waters induces fecal contamination of filter feeding bivalves (oysters, mussels, scallops…), therefore leading to human health risks associated to the consumption of these contaminated organisms. Watershed management plans that aim at limiting these risks require the development of tools able to identify fecal contamination sources. The fecal indicator bacteria used in the regulations to determine fecal contamination are not source specific since they are found in the feces of most warm-blooded animals. Thus, microbiological biomarkers have been developed in association with chemical biomarkers as Microbial Source Tracking (MST) methods. Fecal stanols, by-products of sterols obtained by human and animal microbial gut flora, are found in considerable amounts in feces with different relative proportions depending on their animal or human source. Recently, in association with microbiological biomarkers, the stanol fingerprint of contaminated waters has been successfully used to determine the main source of fecal contamination (cow, pig or human sources) in rural watersheds (Brittany, France). Up to now, the use of the stanol fingerprint to track the fecal contamination in shellfish tissues, especially bivalves, has been limited to the analysis of coprostanol, a stanol commonly associated to human contamination. Therefore, whether the stanol fingerprint can be used as a MST method in bivalves or not is still unknown. The first aim of this study was to compare several organic extraction procedures of stanols in the oyster Crassostrea gigas to determine a reliable method for stanol fingerprint analysis in bivalves. Solvent extraction and purification steps have been carried out with attention as they are critical for stanol quantification. Secondly, the evolution of the stanol fingerprint of oysters with time was evaluated during 6 days by artificially contaminating microcosms with two concentrations of a WWTP effluent. In the microcosms, the fingerprint of stanols as a chemical biomarkers of fecal (human) contamination was compared to counts of Escherichia coli, a commonly used microbial indicator. In association with microbial markers, the method developed from the two previous steps will be applied at the watershed scale in order to identify sources of fecal contamination in Brittany and Normandy (France).
Harrault, Loïc; Jardé, Emilie; Jeanneau, Laurent; Petitjean, Patrice
The bioaccumulation of paralytic shellfish toxins in mussels, oysters, cockles, hard clams, razors, and king scallops is monitored in England, Scotland, and Wales by AOAC Official Method 2005.06 LC-with fluorescence detection (FLD). One of the commonly perceived disadvantages of using this method is the long turnaround time and low throughput in a busy laboratory environment. The chromatographic analysis of each sample typically utilizes a 15 min cycle time to achieve toxin oxidation product separation and column equilibration prior to subsequent analysis. A standard RP C18 analytical column, used successfully in recent years, achieves good separation with a long column lifetime. The analysis of a 40 sample qualitative screening batch takes approximately 18 h, including blanks, standards, and other QC samples. The availability of superficially porous column technology has offered the potential to reduce analysis time while retaining column performance on existing hardware. In this study, AOAC Official Method 2005.06 with LC-FLD was transferred to two different commercially available superficially porous columns, and the method performance characteristics were evaluated. Both columns separated all toxins adequately with cycle times less than half that of the existing method. Linearity for each toxin was acceptable up to two times the European maximum permitted limit of 800 microg di-HCl saxitoxin equivalent/kg flesh. LOD and LOQ values were substantially improved for the majority of toxins, with gonyautoxin 1&4 and neosaxitoxin showing up to a two- and fourfold improvement, respectively, depending on the column used. Quantification results obtained from parallel analysis of contaminated samples were acceptable on both columns. Comparative screen results gave a slight increase in the occurrence of contaminated samples, which was attributed to the improved detection limit for most toxins. Issues with rapidly increasing back pressure, however, were identified with both columns, with a limit of around 500 injections. This compares to the >3000 cycles routinely obtained with the standard RP-C18 HPLC columns currently in use. Overall, the gain achieved with these columns through shorter analysis time and improved analytical sensitivity is potentially of benefit in a high-throughput environment. For the routine high-throughput screening of shellfish samples, however, an improved column lifetime is desirable. PMID:22970577
Hatfield, Robert G; Turner, Andrew D
Fish are routinely exposed to harmful algal blooms that produce noxious compounds and impact the marine food web. This study investigates the role of phase I and II detoxification enzymes on metabolism of the novel paralytic shellfish toxins (PSTs), the hydroxybenzoate analogues recently discovered in Gymnodinium catenatum strains, in the liver of white seabream, assessing ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST) activities. Additionally, the genotoxic potential of hydroxybenzoate PSTs was examined through the erythrocytic nuclear abnormality (ENA) assay. Fish were injected with hydroxybenzoate PSTs into the coelomic cavity and sacrificed 2 and 6 days later for biochemical and cytogenetic analyses. While the activity of EROD was unresponsive to toxins, a significant GST activity decrease was observed at 2 days after injection indicating an impairment of this line of the detoxification system. The genotoxic potential of PSTs was demonstrated by the induction of clastogenic/aneugenic effects at 2 days, as measured by the ENA assay. Overall, this study contributes to better understand the impact of toxins produced by G. catenatum blooms in fish, revealing effects that, even transitory, point out a risk associated to hydroxybenzoate analogues. PMID:22710085
Costa, Pedro Reis; Pereira, Patrícia; Guilherme, Sofia; Barata, Marisa; Santos, Maria Ana; Pacheco, Mário; Pousão-Ferreira, Pedro
Increases in atmospheric carbon dioxide are leading to physical changes in marine environments including parallel decreases in ocean pH and increases in seawater temperature. This study examined the impacts of a six month exposure to combined decreased pH and increased temperature on the immune response and disease status in the blue mussel, Mytilus edulis L. Results provide the first confirmation that exposure to future acidification and warming conditions via aquarium-based simulation may have parallel implications for bivalve health. Collectively, the data suggests that temperature more than pH may be the key driver affecting immune response in M. edulis. Data also suggests that both increases in temperature and/or lowered pH conditions may lead to changes in parasite abundance and diversity, pathological conditions, and bacterial incidence in M. edulis. These results have implications for future management of shellfish under a predicted climate change scenario and future sustainability of shellfisheries. Examination of the combined effects of two stressors over an extended exposure period provides key preliminary data and thus, this work represents a unique and vital contribution to current research efforts towards a collective understanding of expected near-future impacts of climate change on marine environments.
Mackenzie, Clara L.; Lynch, Sharon A.; Culloty, Sarah C.; Malham, Shelagh K.
Okadaic acid and its analogs (OAs) responsible for diarrhetic shellfish poisoning (DSP) strongly inhibit protein phosphatase 2A (PP2A) and thus are quantifiable by measuring the extent of the enzyme inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant human PP2A (rhPP2Ac) for use in a microplate OA assay. OA, dinophysistoxin-1(DTX1), and hydrolyzate of 7-O-palmitoyl-OA strongly inhibited rhPP2Ac activity with IC50 values of 0.095, 0.104, and 0.135 nM, respectively. The limits of detection and quantitation for OA in the digestive gland of scallops and mussels were 0.0348 ?g/g and 0.0611 ?g/g respectively, and, when converted to the whole meat basis, are well below the regulation level proposed by EU (0.16 ?g/g whole meat). A good correlation with LC-MS data was demonstrated, the correlation coefficient being 0.996 with the regression slope of 1.097.
Ikehara, Tsuyoshi; Imamura, Shihoko; Yoshino, Atsushi; Yasumoto, Takeshi
The diarrhetic poisoning by bivalve molluscs, diarrhetic shellfish poisoning, is due to consumption of mussels containing biotoxins produced by some Dinoflagellate species. Toxic effects of yessotoxin (YTX) include morphological alterations of mitochondria from heart and liver but the biochemical basis for these alterations is completely unknown. This paper demonstrates that YTX is a very powerful compound that opens the permeability transition pore (PTP) of the inner mitochondrial membrane of rat liver mitochondria at nanomolar concentrations. The effect requires the presence of a permissive level of calcium, by itself incapable of opening the pore. The direct effect of YTX on PTP is further confirmed by the inhibition exerted by cyclosporin A (CsA) that is known as a powerful inhibitor of PTP opening. Moreover, YTX induces membrane depolarization as shown by the quenching of tetramethylrhodamine methyl ester (TMRM), also prevented by the addition of CsA. YTX caused PTP opening in Morris Hepatoma 1C1 cells, as shown by the occurrence of CsA-sensitive depolarization within minutes of the addition of submicromolar concentrations of the toxin. These results provide a biochemical basis for the mitochondrial alterations observed in the course of intoxication with YTX, offering the first clue into the pathogenesis of diseases caused by YTX, and providing a novel tool to study the PTP in situ. PMID:15178475
Bianchi, Cristina; Fato, Romana; Angelin, Alessia; Trombetti, Fabiana; Ventrella, Vittoria; Borgatti, Anna Rosa; Fattorusso, Ernesto; Ciminiello, Patrizia; Bernardi, Paolo; Lenaz, Giorgio; Parenti Castelli, Giovanna
The production and certification of a series of azaspiracid (AZA) calibration solution reference materials is described. Azaspiracids were isolated from contaminated mussels, purified by preparative liquid chromatography and dried under vacuum to the anhydrous form. The purity was assessed by liquid chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy. The final concentration of each AZA in a CD(3)OH stock solution was determined by quantitative NMR spectroscopy. This solution was then diluted very accurately in degassed, high purity methanol to a concentration of 1.47 ± 0.08 ?mol/L for CRM-AZA1, 1.52 ± 0.05 ?mol/L for CRM-AZA2, and 1.37 ± 0.13 ?mol/L for CRM-AZA3. Aliquots were dispensed into argon-filled glass ampoules, which were immediately flame-sealed. The calibration solutions are suitable for method development, method validation, calibration of liquid chromatography or mass spectrometry instrumentation and quality control of shellfish monitoring programs. PMID:20827466
Perez, Ruth A; Rehmann, Nils; Crain, Sheila; LeBlanc, Patricia; Craft, Cheryl; MacKinnon, Shawna; Reeves, Kelley; Burton, Ian W; Walter, John A; Hess, Philipp; Quilliam, Michael A; Melanson, Jeremy E
The mouse bioassay (MBA) for diarrhetic shellfish poisoning (DSP) toxins has been widely used in many countries of the world. However, different body weight ranges of mice are designated to be used in the Japanese official method and European Union procedure. In this study we investigated whether and to what extent the body weights of the mice affect the susceptibility to DSP toxins. A lethal dose of okadaic acid, one of the representative DSP toxins, was injected intraperitoneally into mice of five different body weight range groups, from 14 to 24 g. The mice were observed until 24 h after injection. The lethality was 100% in the 14-15 and 16-17 g groups, 80% in the 19-20 g group, 50% in the 21-22 g group, and 40% in the 23-24 g group, with significant differences. Survival analysis indicated a relationship between body weights of mice and susceptibility to okadaic acid. These results would be quite useful not only for the MBA, but also to improve understanding of the biological responses to DSP toxins. PMID:24460779
Increases in atmospheric carbon dioxide are leading to physical changes in marine environments including parallel decreases in ocean pH and increases in seawater temperature. This study examined the impacts of a six month exposure to combined decreased pH and increased temperature on the immune response and disease status in the blue mussel, Mytilus edulis L. Results provide the first confirmation that exposure to future acidification and warming conditions via aquarium-based simulation may have parallel implications for bivalve health. Collectively, the data suggests that temperature more than pH may be the key driver affecting immune response in M. edulis. Data also suggests that both increases in temperature and/or lowered pH conditions may lead to changes in parasite abundance and diversity, pathological conditions, and bacterial incidence in M. edulis. These results have implications for future management of shellfish under a predicted climate change scenario and future sustainability of shellfisheries. Examination of the combined effects of two stressors over an extended exposure period provides key preliminary data and thus, this work represents a unique and vital contribution to current research efforts towards a collective understanding of expected near-future impacts of climate change on marine environments. PMID:24927423
Mackenzie, Clara L; Lynch, Sharon A; Culloty, Sarah C; Malham, Shelagh K
This review examines the developments in optical biosensor technology, which uses the phenomenon of surface plasmon resonance, for the detection of paralytic shellfish poisoning (PSP) toxins. Optical biosensor technology measures the competitive biomolecular interaction of a specific biological recognition element or binder with a target toxin immobilised onto a sensor chip surface against toxin in a sample. Different binders such as receptors and antibodies previously employed in functional and immunological assays have been assessed. Highlighted are the difficulties in detecting this range of low molecular weight toxins, with analogues differing at four chemical substitution sites, using a single binder. The complications that arise with the toxicity factors of each toxin relative to the parent compound, saxitoxin, for the measurement of total toxicity relative to the mouse bioassay are also considered. For antibodies, the cross-reactivity profile does not always correlate to toxic potency, but rather to the toxin structure to which it was produced. Restrictions and availability of the toxins makes alternative chemical strategies for the synthesis of protein conjugate derivatives for antibody production a difficult task. However, when two antibodies with different cross-reactivity profiles are employed, with a toxin chip surface generic to both antibodies, it was demonstrated that the cross-reactivity profile of each could be combined into a single-assay format. Difficulties with receptors for optical biosensor analysis of low molecular weight compounds are discussed, as are the potential of alternative non-antibody-based binders for future assay development in this area.
Campbell, K.; Rawn, D.F.K.; Niedzwiadek, B.; Elliott, C.T.
The purpose of the study was to determine if paralytic shellfish poisoning (PSP) toxins are present in extracellular bacteria isolated from a toxic strain of the dinoflagellate Alexandrium minutum. A quantitative analysis was carried out of viable culturable bacteria attached to the surface of dinoflagellates and of bacteria present in dinoflagellate culture medium. A numerical taxonomy study was undertaken for presumptive identification of bacteria attached to the surface of dinoflagellates. Members of the following genera were detected on the cell surface of A. minutum: Cellulophaga, Marinomonas, Pseudoalteromonas, and Vibrio. The presence of intracellular PSP toxins in bacteria isolated from the cell surface of dinoflagellates was analysed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD). Compounds that eluted at the same time as the standards of the PSP toxins GTX-2, GTX-3, GTX-4, dcGTX-2, and dcGTX-3 were present in some of the bacterial cell extracts. Natural fluorescent bacterial compounds, coeluting with some PSP toxins, were also detected. The results obtained showed that the fluorescent compounds, identified as putative PSP toxins by HPLC-FLD, did not correspond to any PSP analogue. This allowed us to reject the hypothesis that extracellular bacteria attached to the surface of dinoflagellates produce PSP toxins. PMID:19898534
Prol, María Jesús; Guisande, Cástor; Barreiro, Aldo; Míguez, Beatriz; de la Iglesia, Pablo; Villar, Adriano; Gago-Martínez, Ana; Combarro, María Pilar
Solid-phase adsorption toxin tracking (SPATT) technology was developed as an effective passive sampling method for dissolved diarrhetic shellfish poisoning (DSP) toxins in seawater. HP20 and SP700 resins have been reported as preferred adsorption substrates for lipophilic algal toxins and are recommended for use in SPATT testing. However, information on the mechanism of passive adsorption by these polymeric resins is still limited. Described herein is a study on the adsorption of OA and DTX1 toxins extracted from Prorocentrum lima algae by HP20 and SP700 resins. The pore size distribution of the adsorbents was characterized by a nitrogen adsorption method to determine the relationship between adsorption and resin porosity. The Freundlich equation constant showed that the difference in adsorption capacity for OA and DTX1 toxins was not determined by specific surface area, but by the pore size distribution in particular, with micropores playing an especially important role. Additionally, it was found that differences in affinity between OA and DTX1 for aromatic resins were as a result of polarity discrepancies due to DTX1 having an additional methyl moiety. PMID:21300360
Li, Aifeng; Ma, Feifei; Song, Xiuli; Yu, Rencheng
The toxicity of the gastropod Nassarius papillosus implicated in a food paralytic poisoning incident in Liuchiu Island, Taiwan, in October 2005 is reported. The symptoms of a victim (67 years old) were featured by general paresthesia, paralysis of phalanges and extremities, paralysis, coma, and aphasia. The remaining specimens of shell were assayed for toxicity. The range of specimen toxicity was found to be 63-474 mouse units (MU) per specimen for N. papillosus by a tetrodotoxin (TTX) bioassay. The mean (SD) toxicity of the digestive gland and other portions were 296 +/- 120 and 382 +/- 156 MU in N. papillosus. The toxin was partially purified from the acidic methanol extract of the gastropod by using a C18 solid-phase extraction column. The eluate was then filtered through a 3000 MW cut-off ultrafree microcentrifuge filter. It was shown that the toxin purified from gastropods analysed by high-performance liquid chromatography and liquid chromatography/mass spectrometry contained TTX 42-60 microg g(-1) (about 90%), whereas along with minor paralytic shellfish poisons (PSP) it was 3-6 microg g(-1) (about 10%). PMID:17613078
Jen, H-C; Lin, S-J; Lin, S-Y; Huang, Y-W; Liao, I-C; Arakawa, O; Hwang, D-F
Many marine bivalve molluscs produce byssal threads for attachment to solid substrata. Small (less than 10 mm) consecutive sections of the byssal threads of Mytilus edulis, M. californianus, Geukensia demissa, Atrina vexillum, and A. rigida were analyzed by amino acid analysis to determine if chemical composition remains constant as a function of location in thread segments. Nonlinear longitudinal protein gradients, probably involving collagen and an elastic protein, were found in the Mytilus species. In these, collagen peaks in the distal third of the thread. In Geukensia and the Atrina species, although the two differed greatly in composition, there is a clear nonvariability in composition of the thread within each species as a function of location in the thread. The adhesive plaque at the tip of the thread of all species examined differs substantially in composition from the remainder of the thread. Protein gradients in the threads of some bivalves may reflect specific adaptations evolved to respond to exposed habitats in high-energy environments. PMID:3095484
Mascolo, J M; Waite, J H
AOAC Official Method 2005.06 precolumn oxidation LC-fluorescence detection method has been used for many years for the detection and quantitation of paralytic shellfish poisoning (PSP) toxins in bivalve molluscs. After extensive single- and multiple-laboratory validation, the method has been slowly gaining acceptance worldwide as a useful and practical tool for official control testing. In Great Britain, the method has become routine since 2008, with no requirement since then for reverting back to the bioassay reference method. Although the method has been refined to be semiautomated, faster, and more reproducible, the quantitation step can be complex and time-consuming. An alternative approach was developed to utilize the qualitative screening results for generating a semiquantitative results assessment. Data obtained over 5 years enabled the comparison of semiquantitative and fully quantitative PSP results in over 15 000 shellfish samples comprising eight different species showed that the semiquantitative approach resulted in over-estimated paralytic shellfish toxin levels by an average factor close to two in comparison with the fully quantified levels. No temporal trends were observed in the data or relating to species type, with the exception of surf clams. The comparison suggested a semiquantitative threshold of 800 microg saxitoxin (STX) eq/kg should provide a safe limit for the determination of samples to be forwarded to full quantitation. However, the decision was taken to halve this limit to include an additional safety factor of 2, resulting in the use of a semiquantitative threshold of 400 microg STX eq/kg. Implementation of the semiquantitative method into routine testing would result in a significant reduction in the numbers of samples requiring quantitation and have a positive impact on the overall turnaround of reported PSP results. The refined method would be appropriate for any monitoring laboratory faced with high throughput requirements. PMID:24830161
Turner, Andrew D; Dhanji-Rapkova, Monika; Baker, Clothilde; Algoet, Myriam
The paralytic shellfish toxins (PSTs) are potent neurotoxic alkaloids and their major biological effect is due to the blockage of voltage-gated sodium channels in excitable cells. They have been recognised as an important health risk for humans, animals, and ecosystems worldwide. The metabolic pathways that lead to the production and the degradation of these toxic metabolites are still unknown. In this study, we investigated the possible link between PST accumulation and the activation of the metabolism that leads to purine degradation in the filamentous freshwater cyanobacterium Planktothrix sp. FP1. The purine catabolic pathway is related to the nitrogen microcycle in water environments, in which cyanobacteria use traces of purines and ureides as a nitrogen source for growth. Thus, the activity of allantoicase, a key inducible enzyme of this metabolism, was used as tool for assaying the activation of the purine degradation pathway. The enzyme and the pathway were induced by allantoic acid, the direct substrate of allantoicase, as well as by adenine and, to a lower degree, by urea, one of the main products of purine catabolism. Crude cell extract of Escherichia coli was also employed and showed the best induction of allantoicase activity. In culture, Planktothrix sp. FP1 showed a differential accumulation of PST in consequence of the induction with different substrates. The cyanobacterial culture induced with allantoic acid accumulated 61.7% more toxins in comparison with the control. On the other hand, the cultures induced with adenine, urea, and the E. coli extract showed low PST accumulation, respectively, 1%, 38%, and 5% of the total toxins content detected in the noninduced culture. A degradation pathway for the PSTs can be hypothesised: as suggested for purine alkaloids in higher plants, saxitoxin (STX) and derivatives may also be converted into xanthine, urea, and further to CO2 and NH4+ or recycled in the primary metabolism through the purine degradation pathway. PMID:11800428
Pomati, F; Manarolla, G; Rossi, O; Vigetti, D; Rossetti, C
Ocean acidification is considered a major threat to marine ecosystems and may particularly affect primary producers. Here we investigated the impact of elevated pCO? on paralytic shellfish poisoning toxin (PST) content and composition in two strains of Alexandrium tamarense, Alex5 and Alex2. Experiments were carried out as dilute batch to keep carbonate chemistry unaltered over time. We observed only minor changes with respect to growth and elemental composition in response to elevated pCO?. For both strains, the cellular PST content, and in particular the associated cellular toxicity, was lower in the high CO? treatments. In addition, Alex5 showed a shift in its PST composition from a non-sulfated analogue towards less toxic sulfated analogues with increasing pCO?. Transcriptomic analyses suggest that the ability of A. tamarense to maintain cellular homeostasis is predominantly regulated on the post-translational level rather than on the transcriptomic level. Furthermore, genes associated to secondary metabolite and amino acid metabolism in Alex5 were down-regulated in the high CO? treatment, which may explain the lower PST content. Elevated pCO? also induced up-regulation of a putative sulfotransferase sxtN homologue and a substantial down-regulation of several sulfatases. Such changes in sulfur metabolism may explain the shift in PST composition towards more sulfated analogues. All in all, our results indicate that elevated pCO? will have minor consequences for growth and elemental composition, but may potentially reduce the cellular toxicity of A. tamarense. PMID:24291633
Van de Waal, Dedmer B; Eberlein, Tim; John, Uwe; Wohlrab, Sylke; Rost, Björn
A large variety of environmental carcinogens are metabolically activated to electrophilic metabolites that can bind to nucleic acids and protein, forming covalent adducts. The formation of DNA-carcinogen adducts is thought to be a necessary step in the action of most carcinogens. Recently, a variety of new fluorescence, immunochemical, and radioactive-postlabeling procedures have been developed that allow the sensitive measurement of DNA-carcinogen adducts in organisms exposed to environmental carcinogens. In some cases, similar procedures have been developed for protein-carcinogen adducts. In an organism with active metabolic systems for a given carcinogen, adducts are generally much longer lived than the carcinogens that formed them. Thus, the detection of DNA- or protein-carcinogen adducts in aquatic foodstuffs can act as an indicator of prior carcinogen exposure. The presence of DNA adducts would, in addition, suggest a mutagenic/carcinogenic risk to the aquatic organism itself. Vertebrate fish are characterized by high levels of carcinogen metabolism, low body burdens of carcinogen, the formation of carcinogen-macromolecule adducts, and the occurrence of pollution-related tumors. Shellfish, on the other hand, have low levels of carcinogen metabolism, high body burdens of carcinogen, and have little or no evidence of carcinogen-macromolecule adducts or tumors. The consumption of carcinogen adducts in aquatic foodstuffs is unlikely to represent a human health hazard. There are no metabolic pathways by which protein-carcinogen or DNA-carcinogen adducts could reform carcinogens. Incorporation via salvage pathways of preformed nucleoside-carcinogen adducts from foodstuffs into newly synthesized human DNA is theoretically possible.(ABSTRACT TRUNCATED AT 250 WORDS) Images FIGURE 1. FIGURE 1. FIGURE 2.
Dunn, B P
The receptor-binding assay (RBA) method for determining saxatoxin (STX) and its numerous analogues, which cause paralytic shellfish poisoning (PSP) in humans, was evaluated in a single laboratory study. Each step of the assay preparation procedure including the performance of the multi-detector TopCount® instrument was evaluated for its contribution to method variability. The overall inherent RBA variability was determined to be 17%. Variability within the 12 detectors was observed; however, there was no reproducible pattern in detector performance. This observed variability among detectors could be attributed to other factors, such as pipetting errors. In an attempt to reduce the number of plates rejected due to excessive variability in the method's quality control parameters, a statistical approach was evaluated using either Grubbs' test or the Student's t-test for rejecting outliers in the measurement of triplicate wells. This approach improved the ratio of accepted versus rejected plates, saving cost and time for rerunning the assay. However, the potential reduction in accuracy and the lack of improvement in precision suggests caution when using this approach. The current study has recommended an alternate quality control procedure for accepting or rejecting plates in place of the criteria currently used in the published assay, or the alternative of outlier testing. The recommended procedure involves the development of control charts to monitor the critical parameters identified in the published method (QC sample, EC??, slope of calibration curve), with the addition of a fourth critical parameter which is the top value (100% binding) of the calibration curve. PMID:22928711
Ruberu, S R; Langlois, G W; Masuda, M; Perera, S Kusum
The temporal dynamics of carbon and nitrogen isotope values of co-occurring suspension-feeders in two shellfish culture areas (Normandy, France) were investigated over two years to evaluate the inter-specific trophic partitioning and relative contributions of organic matter sources to benthic suspension-feeders' diet. Oysters ( Crassostrea gigas), mussels ( Mytilus edulis), cockles ( Cerastoderma edule), slipper limpets ( Crepidula fornicata), and sand-mason worms ( Lanice conchilega) were sampled in an estuarine environment (Baie des Veys, east Cotentin, Normandy), while oysters, mussels, slipper limpets, and honeycomb worms ( Sabellaria alveolata) were sampled in an open-marine environment (Lingreville-sur-mer, west Cotentin, Normandy). Whatever the sampling period, the bivalves, C. gigas and M. edulis, exhibited the lowest values of ?13C and ?15N compared with the other species. Feeding relationships among suspension-feeders in both C. gigas culture areas exhibited temporal variations due to the marine/estuarine influence and seasonal changes in food supply. In the open-marine ecosystem, the contribution of phytoplankton remained the most important for all species except S. alveolata, while in the estuarine ecosystem, microphytobenthos and/or macroalgae detritus contributed a larger extent to the organisms' diets. During phytoplankton bloom periods (e.g. May and July) suspension-feeders, except for S. alveolata, relied strongly on phytoplankton; however, the majority of suspension-feeders exhibited different opportunistic behaviour in winter when phytoplankton biomass might be a limiting factor. We hypothesized that differences in particle capture and selection by the suspension-feeders influenced their isotopic values. Feeding ecology of suspension-feeders partly explained why competition was limited and why ecosystems can often support unexpectedly large numbers of suspension-feeders. We also showed that understanding ecosystem characteristics of the organic matter sources is of primary importance to determine the extent to which members of the suspension-feeding guild potentially compete for food.
Lefebvre, Sébastien; Marín Leal, Julio César; Dubois, Stanislas; Orvain, Francis; Blin, Jean-Louis; Bataillé, Marie-Paule; Ourry, Alain; Galois, Robert
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. Images
Chen, D; Hanna, P J; Altmann, K; Smith, A; Moon, P; Hammond, L S
Fish are recurrently exposed to paralytic shellfish toxins (PSTs) produced by Gymnodinium catenatum. Nevertheless, the knowledge regarding metabolism of PSTs and their toxic effects in fish is scarce. Consequently, the current study aims to investigate the role of phase I and II detoxification enzymes on PST metabolism in the liver of white seabream (Diplodus sargus), assessing ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST) activities. Moreover, the genotoxic potential of PSTs was examined through the erythrocytic nuclear abnormality (ENA) assay. Fish were intracoelomically (IC) injected with a nominal dose (expressed as saxitoxin equivalents) of 1.60 ?g STXeq kg?¹ semipurified from a G. catenatum cell culture with previously determined toxin profile. Fish were sacrificed 2 and 6 days after IC injection. PST levels determined in fish liver were 15.2 and 12.2 ?g STXeq kg?¹, respectively, at 2 and 6 days after the injection. Though several PSTs were administered, only dcSTX was detected in the liver after 2 and 6 days. This was regarded as an evidence that most of the N-sulfocarbamoyl and decarbamoyl toxins were rapidly biotransformed in D. sargus liver and/or eliminated. This was corroborated by a hepatic GST activity induction at 2 days after injection. Hepatic EROD activity was unresponsive to PSTs, suggesting that these toxins enter phase II of biotransformation directly. The genotoxic potential of PSTs was also demonstrated; these toxins were able to induce cytogenetic damage, such as chromosome (or chromatid) breaks or loss and segregational anomalies, measured by the ENA assay. Overall, this study pointed out the ecological risk associated with the contamination of fish with PSTs generated by G. catenatum blooms, providing the necessary first data for a proper interpretation of biomonitoring programs aiming to assess the impact of phytoplankton blooms in fish. PMID:22057254
Costa, Pedro Reis; Pereira, Patrícia; Guilherme, Sofia; Barata, Marisa; Nicolau, Lídia; Santos, Maria Ana; Pacheco, Mário; Pousão-Ferreira, Pedro
Farmed greenlip abalone Haliotis laevigata were fed commercial seaweed-based food pellets or feed pellets supplemented with 8 × 10? Alexandrium minutum dinoflagellate cells g?¹ (containing 12 ± 3.0 ?g STX-equivalent 100 g?¹, which was mainly GTX-1,4) every second day for 50 days. Exposure of abalone to PST supplemented feed for 50 days did not affect behaviour or survival but saw accumulation of up to 1.6 ?g STX-equivalent 100 g?¹ in the abalone foot tissue (muscle, mouth without oesophagus and epipodial fringe), which is ?50 times lower than the maximum permissible limit (80 ?g 100 g?¹ tissue) for PSTs in molluscan shellfish. The PST levels in the foot were reduced to 0.48 ?g STX-equivalent 100 g?¹ after scrubbing and removal of the pigment surrounding the epithelium of the epipodial fringe (confirmed by both HPLC and LC-MS/MS). Thus, scrubbing the epipodial fringe, a common procedure during commercial abalone canning, reduced PST levels by ?70%. Only trace levels of PSTs were detected in the viscera (stomach, gut, heart, gonad, gills and mantle) of the abalone. A toxin reduction of approximately 73% was observed in STX-contaminated abalone held in clean water and fed uncontaminated food over 50 days. The low level of PST uptake when abalone were exposed to high numbers of A. minutum cells over a prolonged period may indicate a low risk of PSP poisoning to humans from the consumption of H. laevigata that has been exposed to a bloom of potentially toxic A. minutum in Australia. Further research is required to establish if non-dietary accumulation can result in significant levels of PSTs in abalone. PMID:21640130
Dowsett, Natalie; Hallegraeff, Gustaaf; van Ruth, Paul; van Ginkel, Roel; McNabb, Paul; Hay, Brenda; O'Connor, Wayne; Kiermeier, Andreas; Deveney, Marty; McLeod, Catherine
Uptake and depuration of paralytic shellfish toxins in the green-lipped mussel, Perna viridis, were investigated by exposing the mussels to dinoflagellates (Alexandrium tamarense, ACTI01) under laboratory conditions for 8 d, then depurating them in clean seawater for 14 d. First-order linear differential equations were set up for five tissue compartments: Viscera, gill, hepatopancreas, adductor muscle, and foot. The solutions to these equations were used to fit the experimental data. We then estimated the parameters governing the model, which depend on the elimination rate from each compartment and the transfer coefficient between compartments. An assumption of the model is that the gills transport the dinoflagellates directly to the mouth and then to the viscera, where the ingested cells are broken down, releasing the toxins. The toxins absorbed are transferred to other tissues. During the uptake phase, the transfer coefficients from viscera to gill, hepatopancreas, adductor muscle, and foot were 0.03, 0.24, 0.01, and 0.004 per day, respectively. During the depuration phase, the transfer coefficients were 0.01, 0, 0.01, and 0.003 per day, respectively. In terms of the anatomical distribution of N-sulfocarbamoyl-11-hydroxysulfate (C2) toxins in various tissues, viscera and hepatopancreas contained the highest percentages (47-74% and 8-41%, respectively). Together, these two tissue compartments accounted for 71 to 96% of all C2 toxins present. The biokinetic model allows a quantitative prediction of C2 toxins in whole ussel as well as individual tissue compartments based on the density estimates and toxin load of dinoflagellate cells in the surrounding waters over time. PMID:15683176
Li, Ashley M Y; Yu, Peter K N; Hsieh, Dennis; Wang, Wen-Xiong; Wu, Rudolf S S; Lam, Paul K S
We exposed green-lipped mussels Perna viridis and black sea breams Acanthopagrus schlegeli to toxic dinoflagellates Alexandrium fundyense to evaluate the accumulation, distribution, transformation, and elimination of paralytic shellfish toxins (PSTs) in a controlled environmental condition. The mussels were fed A. fundyense for 7 days followed by 3 weeks of depuration, and the fish were fed toxic clams (pre-exposed to the dinoflagellates) for 5 days followed by 2 weeks of depuration. The toxin content and the compartmental distribution of PSTs were monitored throughout the experiments by high-performance liquid chromatography with post-column fluorescence derivatization (HPLC-FLD). This is the first report to assess the biokinetics of PSTs in marine fish under dietary exposure. The hepatopancreas in the mussels and the viscera in the fish accumulated most of the PSTs. Differential elimination of each toxin was observed in the mussels. The C2 toxins were eliminated rapidly in all organs; except in hepatopancreas, the more potent toxins such as GTX4, were eliminated slower during the depuration period. The relative proportions of various PSTs in the mussels changed over time, suggesting toxin-specific uptake and elimination rates, or biotransformation preferences between toxins. In the fish, the ratio of C1/C2 was 3.0 times (p<0.01) higher when compared to the clam tissues, indicating that conversion from C2 to C1 might have occurred when the toxin was transferred from the clams to the fish. In summary, species differences in uptake, distribution and elimination of PSTs were observed between mussels and fish, and this may influence trophic transfer of algal toxins in marine organisms. PMID:16959334
Kwong, Raymond W M; Wang, Wen-Xiong; Lam, Paul K S; Yu, Peter K N
In this paper we describe optimization of SYBR Green I-based real-time PCR parameters and testing of a large number of microbial species with vvh-specific oligonucleotide primers to establish a rapid, specific, and sensitive method for detection of Vibrio vulnificus in oyster tissue homogenate and Gulf of Mexico water (gulf water). Selected oligonucleotide primers for the vvh gene were tested for PCR amplification of a 205-bp DNA fragment with a melting temperature of approximately 87°C for 84 clinical and environmental strains of V. vulnificus. No amplification was observed with other vibrios or nonvibrio strains with these primers. The minimum level of detection by the real-time PCR method was 1 pg of purified genomic DNA or 102 V. vulnificus cells in 1 g of unenriched oyster tissue homogenate or 10 ml of gulf water. It was possible to improve the level of detection to one V. vulnificus cell in samples that were enriched for 5 h. The standard curves prepared from the real-time PCR cycle threshold values revealed that there was a strong correlation between the number of cells in unenriched samples and the number of cells in enriched samples. Detection of a single cell of V. vulnificus in 1 g of enriched oyster tissue homogenate is in compliance with the recent Interstate Shellfish Sanitation Conference guidelines. The entire detection method, including sample processing, enrichment, and real-time PCR amplification, was completed within 8 h, making it a rapid single-day assay. Rapid and sensitive detection of V. vulnificus would ensure a steady supply of postharvest treated oysters to consumers, which should help decrease the number of illnesses or outbreaks caused by this pathogen.
Panicker, Gitika; Myers, Michael L.; Bej, Asim K.
Vibrio parahaemolyticus is an important human pathogen which can cause gastroenteritis when consumed in raw or partially-cooked seafood. A multiplex PCR amplification-based detection of total and virulent strains of V. parahaemolyticus was developed by targeting thermolabile hemolysin encoded by tl, thermostable direct hemolysin encoded by tdh, and thermostable direct hemolysin-related trh genes. Following optimization using oligonucleotide primers targeting tl, tdh and trh genes, the multiplex PCR was applied to V. parahaemolyticus from 27 clinical, 43 seafood, 15 environmental, 7 strains obtained from various laboratories and 19 from oyster plants. All 111 V. parahaemolyticus isolates showed PCR amplification of the tl gene; however, only 60 isolates showed amplification of tdh, and 43 isolates showed amplification of the trh gene. Also, 18 strains showed amplification of the tdh gene, but these strains did not show amplification of the trh gene. However, one strain exhibited amplification for the trh but not the tdh gene, suggesting both genes need to be targeted in a PCR amplification reaction to detect all hemolysin-producing strains of this pathogen. The multiplex PCR approach was successfully used to detect various strains of V parahaemolyticus in seeded oyster tissue homogenate. Sensitivity of detection for all three target gene segments was at least between 10(1)-10(2) cfu per 10 g of alkaline peptone water enriched seeded oyster tissue homogenate. This high level of sensitivity of detection of this pathogen within 8 h of pre-enrichment is well within the action level (10(4) cfu per 1 g of shell stock) suggested by the National Seafood Sanitation Program guideline. Compared to conventional microbiological culture methods, this multiplex PCR approach is rapid and reliable for accomplishing a comprehensive detection of V. parahaemolyticus in shellfish. PMID:10379807
Bej, A K; Patterson, D P; Brasher, C W; Vickery, M C; Jones, D D; Kaysner, C A
A lateral flow immunoassay (LFIA) has been developed and fully validated to detect the primary amnesic shellfish poisoning (ASP) toxin, domoic acid (DA). The performance characteristics of two versions of the test were investigated using spiked and naturally contaminated shellfish (mussels, scallops, oysters, clams, and cockles). The tests provide a qualitative result, to indicate the absence or presence of DA in extracts of shellfish tissues, at concentrations that are relevant to regulatory limits. The new rapid assay (LFIA version 2) was designed to overcome the performance limitations identified in the first version of the assay. The improved test uses an electronic reader to remove the subjective nature of the generated results, and the positive cut-off for screening of DA in shellfish was increased from 10 ppm (version 1) to 17.5 ppm (version 2). A simple extraction and test procedure was employed, which required minimal equipment and materials; results were available 15 min after sample preparation. Stability of the aqueous extracts at room temperature (22 °C) at four time points (up to 245 min after extraction) and across a range of DA concentrations was 100.3±1.3% and 98.8±2.4% for pre- and post-buffered extracts, respectively. The assay can be used both within laboratory settings and in remote locations. The accuracy of the new assay, to indicate negative results at or below 10 ppm DA, and positive results at or above 17.5 ppm, was 99.5% (n=216 tests). Validation data were obtained from a 2-day, randomised, blind study consisting of multiple LFIA lots (n=3), readers (n=3) and operators (n=3), carrying out multiple extractions of mussel tissue (n=3) at each concentration (0, 10, 17.5, and 20 ppm). No matrix effects were observed on the performance of the assay with different species (mussels, scallops, oysters, clams, and cockles). There was no impact on accuracy or interference from other phycotoxins, glutamic acid or glutamine with various strip incubations (8, 10, and 12 min). The accuracy of the assay, using naturally contaminated samples to indicate negative results at or below 12.5 ppm and positive results at or above 17.5 ppm, was 100%. Variability between three LFIA lots across a range of DA concentrations, expressed as coefficient of variation (% CV), was 1.1±0.4% (n=2 days) based on quantitative readings from the electronic reader. During an 8 week stability study, accuracy of the method with test strips stored at various temperatures (6, 22, 37 and 50 °C) was 100%. Validation for both versions included comparisons with results obtained using reference LC-UV methods. PMID:24148459
Jawaid, Waqass; Meneely, Julie; Campbell, Katrina; Hooper, Mark; Melville, Karrie; Holmes, Stephen; Rice, Jennifer; Elliott, Christopher
LC/MS methodology for the analysis of domoic acid and lipophilic toxins in shellfish was developed using a hybrid triple quadrupole linear ion trap mass spectrometer. For routine quantitation a scheduled selected reaction monitoring (SRM) method for the analysis of domoic acid, okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, gymnodimines, spirolides, and pinnatoxins was developed and validated. The method performed well in terms of LOD, linearity, precision, and trueness. Taking advantage of the high instrument sensitivity, matrix effects were mitigated by reducing the amount of sample introduced to the mass spectrometer. Optionally, samples can be analyzed using information dependent acquisition (IDA) methods, either in positive or negative mode, which can provide an extra level of confirmation by matching the full product ion spectra acquired for a sample with those from a specially constructed spectral library. Methods were applied to the analysis df a new certified reference material and Canadian mussels (Mytilus edulis) implicated in a 2011 diarrhetic shellfish poisoning (DSP) incident. The scheduled SRM method enabled the screening and quantitation of multiple phycotoxins. As DSP had not previously been observed in this area of Canada, positive identification of putative toxins was accomplished using the IDA and spectral search method. Analysis of the 2011 toxic mussel samples revealed the presence of high levels of dinophysistoxin-1, which explained the DSP symptoms, as well as pectenotoxins, yessotoxins, and variety of cyclic imines. PMID:24830142
McCarron, Pearse; Wright, Elliott; Quilliam, Michael A
The objective of the Hanford Environmental Dose Reconstruction Project (HEDR) is to estimate the potential radiation doses received by people living within the sphere of influence of the Hanford Site. A potential critical pathway for human radiation exposure is through the consumption of waterfowl that frequent onsite waste-water ponds or through eating of fish, shellfish, and waterfowl that reside in/on the Columbia River and its tributaries downstream of the reactors. This document summarizes information on fish, shellfish, and waterfowl radiation contamination for samples collected by Hanford monitoring personnel and offsite agencies for the period 1945 to 1972. Specific information includes the types of organisms sampled, the kinds of tissues and organs analyzed, the sampling locations, and the radionuclides reported. Some tissue concentrations are also included. We anticipate that these yearly summaries will be helpful to individuals and organizations interested in evaluating aquatic pathway information for locations impacted by Hanford operations and will be useful for planning the direction of future HEDR studies.
Hanf, R.W.; Dirkes, R.L.; Duncan, J.P.
This work had as main objectives to characterize two bacteriocins produced by lactic acid bacteria (LAB) previously isolated from non-fermented seafood, in order to evaluate their potential as new food protective agents. The two bacteriocinogenic isolates were identified by Polymerase Chain Reaction (PCR) using genus- and species-specific primers, and confirmed by 16S rDNA sequencing, as Enterococcus faecium and Pediococcus pentosaceus. The antimicrobial spectrum of each strain included several indicator microorganisms, some of them also isolated from seafood. Growth of Listeria innocua, L. monocytogenes, Staphylococcus aureus, Bacillus cereus and other LAB species were inhibited, although no inhibition of Gram-negative microorganisms was observed. Proteolytic, but not lipolytic or glycolytic enzymes, completely inactivated the antimicrobial effect of both cell-free supernatants confirming the proteinaceous nature of the inhibitors. The antimicrobial activity was maintained after treatment with NaCl, SDS, Triton X-100, Tween 20, Tween 80 and EDTA after 2 h or 5 h of exposure and both bacteriocins were stable over a wide range of pH and temperatures. Production of bacteriocin by E. faecium (bacALP7) was detected initially at exponential phase and reached a maximum activity of 25,600 AU/ml in the early stationary phase, whereas bacteriocin production by P. pentosaceus ALP57 (bacALP57) reached the maximum at exponential phase with 12,800 AU/ml. The bacteriocins did not kill L. monocytogenes ESB54 nor L. innocua 2030c however, cellular growth was reduced. The partially purified bacteriocins, bacALP7 and bacALP57, were below 6.5 kDa in size as determined by Tricine-SDS gel electrophoresis. E. faecium and P. pentosaceus contained DNA fragments corresponding in size to those recorded for enterocin B and pediocin PA-1, respectively. Sequencing of the fragments from both bacteriocins confirmed the homology. To our knowledge, for the first time two LAB producing bacteriocins similar to pediocin PA-1 and enterocin B, were isolated from non-fermented shellfish. The adaptation of the cultures to seafood matrices may be advantageous in terms of application as a biopreservation strategy for reduction of L. monocytogenes levels in seafood products. PMID:19081155
Pinto, Ana Luísa; Fernandes, Melissa; Pinto, Cristina; Albano, Helena; Castilho, Fernanda; Teixeira, Paula; Gibbs, Paul A
Significant differences previously observed in the determination of paralytic shellfish poisoning toxins (PSTs) in oysters using official method AOAC 2005.06 and 959.08 were investigated in detail with regard to possible matrix effects. Method AOAC 2005.06 gave results 2-3 times higher than the mouse bioassay method, 959.08, differences thought to be due to underestimation of PSTs by the mouse bioassay. In order to prove the cause of these large differences, work was conducted here to examine the presence and effects of matrix components on the performance of each of the two assays. A range of oyster, cockle and mussel samples were extracted using the AOAC 959.08 hydrochloric acid (HCl) extraction method and analysed for PSP by both MBA and LC-FLD. In addition, extracts were analysed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) for metals as well as being subjected to a range of nutritional testing methods. Whilst there was no evidence for effect of nutritional components on either assay, ICP-MS analysis revealed a relationship between samples exhibiting the largest differences in relative method performance, specifically those with the largest LC-FLD/MBA toxicity ratio, and samples containing the highest concentrations of zinc and manganese. In order to prove the potential effect of the metals on either the LC-FLD and/or MBA assays, HCl extracts of a range of shellfish were subjected to a number of matrix modifications. Firstly, a number of PSP-positive oyster samples were processed to reduce the concentrations of metals within the extracts, without significantly reducing the concentrations of PSTs. Secondly, a range of mussel and cockle extracts, plus a standard solution of saxitoxin di-hydrochloride were spiked at variable concentrations of zinc. All treated and non-treated extracts, plus a number of controls were subjected to ICP-MS, LC-FLD and MBA testing. Results proved the absence of any effect of metals on the performance of the LC-FLD, whilst showing a large suppressive effect of the metals on the MBA. As such, the results show the performance of the official MBA is potentially unsafe for application to the routine monitoring of PSP toxicity in oysters or in any other shellfish found to contain high concentrations of metal ions. PMID:22138287
Turner, Andrew D; Dhanji-Rapkova, Monika; Algoet, Myriam; Suarez-Isla, Benjamin A; Cordova, Marco; Caceres, Catherine; Murphy, Cory J; Casey, Melanie; Lees, David N
Located in Bivalve, New Jersey, HSRL has over a 100-yr tradition of disseminating research results and working cooperatively with state and federal agencies and the fisheries and aquaculture communities in southern New Jersey. Site features information on the lab history, facilities, faculty, and current and past research. An abundance of information regarding oyster and clam aquaculture, as well as links to additional resources.
... monitoring programs (assessing toxin levels in mussels, oysters, scallops, clams) and rapid closures of suspect or demonstrated ... of North America Mussels, surfclams, softshell clams, sea scallops, butterclams, ocean quahogs, oysters, gastropods, lobsters, crabs. Herring, ...
A closed vessel microwave digestion procedure was developed for shellfish samples. This procedure was compared with wet and dry ash procedures for levels of lead, cadmium, chromium, copper, and zinc. Results obtained for microwave and conventional wet ash digestion were comparable. The dry ashing procedure produced results consistently lower than either of the other methods. Recoveries ranged from 80-92% for microwave and wet ashing procedures and 54-72% for the dry ashing procedure. Accuracy was also determined by analyzing lobster hepatopancreas marine reference material. Values for Pb, Cd, and Cr fell within the range specified for the reference material for all 3 digestion procedures; however, values were lower for Cu and Zn. Results of this study show that microwave digestion is comparable to wet ashing. PMID:1874704
McCarthy, H T; Ellis, P C
The increased frequency and distribution of red tides requires the development of high-throughput detection methods for paralytic shellfish toxins (PST). Community ethics also requires that there be a reduced reliance upon the standard mouse bioassay. A biomolecular assay such as the sodium channel 3H-saxitoxin binding assay can satisfy both of these requirements but may be compromised by cross-reactivity with the
Andrew Negri; Lyndon Llewellyn
Thin-layer chromatography (TLC) on Chromarods-SIII with the Iatroscan (Mark-5) and a flame thermionic detector (FTID) was used to develop a rapid method for the detection of paralytic shellfish poisoning (PSP) toxins. The effect of variation in hydrogen (H2) flow, air flow, scan time and detector current on the FTID peak response for both phosphatidylcholine (PC) and PSP were studied in order to define optimum detection conditions. A combination of hydrogen and air flow-rates of 50 ml/min and 1.5-2.0 l/min respectively, along with a scan time of 40 s/rod and detector current of 3.0 A (ampere) or above were found to yield the best results for the detection of PSP compounds. Increasing the detector current level to as high as 3.3 A gave about 130 times more FTID response than did flame ionization detection (FID), for PSP components. Quantities of standards as small as 1 ng neosaxitoxin (NEO), 5 ng saxitoxin (STX), 5 ng B1-toxins (B1), 2 ng gonyautoxin (GTX) 2/3, 6 ng GTX 1/4 and 6 ng C-toxins (C1/C2) could be detected with the FTID. The method detection limits for toxic shellfish tissues using the FTID were 0.4, 2.1, 0.8 and 2.5 micrograms per g tissue for GTX 2/3, STX, NEO and C toxins, respectively. The FTID response increased with increasing detector current and with increasing the scan time. Increasing hydrogen and air flow-rates resulted in decreasing sensitivity within defined limits. Numerous solvent systems were tested, and, solvent consisting of chloroform: methanol-water-acetic acid (30:50:8:2) could separate C toxins from GTX, which eluted ahead of NEO and STX. Accordingly, TLC/FTID with the Iatroscan (Mark-5) seems to be a promising, relatively inexpensive and rapid method of screening plant and animal tissues for PSP toxins. PMID:10519102
Indrasena, W M; Ackman, R G; Gill, T A
The marine environment in Kuwait is dominated by Kuwait Bay, a shallow, depositional habitat vital for the breeding and propagation of marine organisms. The bay receives effluent inputs from industrial centres, ports, sewage outflows along with discharges from power and desalination plants. The major classes of pollutant discharged into the bay include petroleum hydrocarbons, metals, nutrients, cooling water and hyper-saline water. Further, the bay has been historically impacted by a deliberate release of oil and contamination with ordnance and shipwrecks during the 1991 Gulf war. With an aim to establish an integrated pollution effects monitoring programme in Kuwait, this paper describes the application of a quality assured approach to conduct a histopathology baseline survey in oriental sole (Synaptura orientalis) and the large-toothed flounder (Pseudorhombus arsius), which are two potential sentinel flatfish species present in the Arabian Gulf. Liver and gonadal histopathology revealed a range of pathologies similar to those previously observed in European and American pollution effects surveys that utilise flatfish (including pathology markers indicative of possible carcinogenesis and endocrine disruption). Further, we extended these studies to invertebrates (Jinga prawn, Metapenaeus affinis and the grooved tiger prawn, Penaeus semisulcatus) found within the Arabian Gulf. Such baseline data is essential before attempts are made to develop integrated monitoring programmes that aim to assess the health of fish and shellfish in relation to chemical contamination. PMID:24680107
Stentiford, G D; Massoud, M S; Al-Mudhhi, S; Al-Sarawi, M A; Al-Enezi, M; Lyons, B P
First evidence of "paralytic shellfish toxins" and cylindrospermopsin in a Mexican freshwater system, Lago Catemaco, and apparent bioaccumulation of the toxins in "tegogolo" snails (Pomacea patula catemacensis).
Exposure to cyanobacterial toxins in freshwater systems, including both direct (e.g., drinking water) and indirect (e.g., bioaccumulation in food webs) routes, is emerging as a potentially significant threat to human health. We investigated cyanobacterial toxins, specifically cylindrospermopsin (CYN), the microcystins (MCYST) and the "paralytic shellfish toxins" (PST), in Lago Catemaco (Veracruz, Mexico). Lago Catemaco is a tropical lake dominated by Cylindrospermopsis, specifically identified as Cylindrospermopsis catemaco and Cylindrospermopsis philippinensis, and characterized by an abundant, endemic species of snail (Pomacea patula catemacensis), known as "tegogolos," that is both consumed locally and commercially important. Samples of water, including dissolved and particulate fractions, as well as extracts of tegogolos, were screened using highly specific and sensitive ELISA. ELISA identified CYN and PST at low concentrations in only one sample of seston; however, both toxins were detected at appreciable quantities in tegogolos. Calculated bioaccumulation factors (BAF) support bioaccumulation of both toxins in tegogolos. The presence of CYN in the phytoplankton was further confirmed by HPLC-UV and LC-MS, following concentration and extraction of algal cells, but the toxin could not be confirmed by these methods in tegogolos. These data represent the first published evidence for CYN and the PST in Lago Catemaco and, indeed, for any freshwater system in Mexico. Identification of the apparent bioaccumulation of these toxins in tegogolos may suggest the need to further our understanding of the transfer of cyanobacterial toxins in freshwater food webs as it relates to human health. PMID:19651152
Berry, John P; Lind, Owen
The objective of this work was to study the effects of washing and purification steps on qualitative and quantitative analysis of fecal stanols in the oyster Crassostrea gigas using either single or a combination of lipid purification steps on silica gel or aminopropyl bonded silica gel (NH2) or a washing step. Among the three analytical pathways compared, the two including water extraction or NH2 purification did not lead to higher recoveries and decreased repeatabilities of extractions compared to the single purification on silica gel. This latter led to similar recoveries (ca. 80 %) and repeatabilities (ca. 10 %) for both spiked standards (coprostanol and sitostanol). This analytical pathway has been applied to oysters collected in a harvesting area in Brittany (France) where fecal contaminations are important and allowed to quantify eight stanols in oysters. The relative proportions of fecal stanols of these oysters were combined with principal component analysis in order to investigate the usefulness of their stanol fingerprints to record a fecal contamination and to distinguish its source between human, porcine and bovine contaminations. Oysters non-fecally contaminated by Escherichia coli did not present specific stanol fingerprints while oysters fecally contaminated had a bovine fingerprint, suggesting a contamination of these samples by bovine sources. As a consequence, the method developed here allows the use of stanol fingerprints of oysters as a microbial source tracking tool that can be applied to shellfish harvesting areas subjected to fecal contaminations in order to identify the different sources of contamination and improve watershed management. PMID:24771549
Harrault, Loïc; Jardé, Emilie; Jeanneau, Laurent; Petitjean, Patrice
Humans have contributed to phenotypic and demographic changes in their prey from very early on in the colonization of Europe, including the harvesting of shellfish in coastal ecosystems. We estimated trends in population growth (variation in the number of individuals) from DNA sequences of modern specimens in two North Iberian molluscs, top shells (Osilinus lineatus, from 24 sequences and 14 haplotypes) and limpets (Patella vulgata, taken from the bibliography), which were subjected to very different levels of harvesting pressure during the Upper Palaeolithic (~ 20000 to ~ 6000 years ago). The less harvested Osilinus top shells experienced fluctuations in population numbers coincident with climatic oscillations. Patella limpets, which were harvested in greater numbers, suffered clear and uninterrupted decreases in their numbers during the Upper Palaeolithic. These trends coincided with morphological changes in shell size (length or width) in the same direction (i.e., shell size decreased when population size decreased and vice versa). The differing trends seen in taxa subjected to different intensities of harvesting pressure suggest that climate effects were overcome by anthropogenic selection (leading to a smaller average length) in limpets. We suggest that intense fishing pressure may have induced irreversible shell length decreases in the most exploited species.
Turrero, Pablo; Muñoz-Colmenero, A. Marta; Prado, Andrea; Garcia-Vazquez, Eva
Confirmation of Pinnatoxins and Spirolides in Shellfish and Passive Samplers from Catalonia (Spain) by Liquid Chromatography Coupled with Triple Quadrupole and High-Resolution Hybrid Tandem Mass Spectrometry
Cyclic imines are lipophilic marine toxins that bioaccumulate in seafood. Their structure comprises a cyclic-imino moiety, responsible for acute neurotoxicity in mice. Cyclic imines have not been linked yet to human poisonings and are not regulated in Europe, although the European Food Safety Authority requires more data to perform a conclusive risk assessment for consumers. This work presents the first detection of pinnatoxin G (PnTX-G) in Spain and 13-desmethyl spirolide C (SPX-1) in shellfish from Catalonia (Spain, NW Mediterranean Sea). Cyclic imines were found at low concentrations (2 to 60 µg/kg) in 13 samples of mussels and oysters (22 samples analyzed). Pinnatoxin G has been also detected in 17 seawater samples (out of 34) using solid phase adsorption toxin tracking devices (0.3 to 0.9 µg/kg-resin). Pinnatoxin G and SPX-1 were confirmed with both low and high resolution (<2 ppm) mass spectrometry by comparison of the response with that from reference standards. For other analogs without reference standards, we applied a strategy combining low resolution MS with a triple quadrupole mass analyzer for a fast and reliable screening, and high resolution MS LTQ Orbitrap® for unambiguous confirmation. The advantages and limitations of using high resolution MS without reference standards were discussed.
Garcia-Altares, Maria; Casanova, Alexis; Bane, Vaishali; Diogene, Jorge; Furey, Ambrose; de la Iglesia, Pablo
The cyanotoxin, microcystin (MC), is known to accumulate in the tissues of diverse aquatic biota although factors influencing exposure, such as feeding habits and seasonal patterns in toxin production, are poorly known. We analyzed seasonal variation in the MC content of primary and secondary consumers, and used dietary analysis (gut contents and stable isotopes) to improve understanding of cyanotoxin transport in food webs. Periods of elevated toxin concentration were associated with peaks in the abundance of genes specific to Microcystis and MC toxin production (mcyD). Peak toxin levels in consumer tissues coincided with peak MC concentrations in seston. However, toxins in tissues persisted in overwintering populations suggesting that potential health impacts may not be limited to bloom periods. Interspecific differences in tissue MC concentrations were related to feeding habits and organic matter sources as pelagic fishes ingested a greater proportion of algae in their diet, which resulted in greater MC content in liver and muscle tissues. Sediments contained a greater proportion of allochthonous (terrestrial) organic matter and lower concentrations of MC, resulting in lower toxin concentrations among benthic detritivores. Among shellfish, the benthic suspension feeder Rangia cuneata (wedge clam) showed seasonal avoidance of toxin ingestion due to low feeding rates during periods of elevated MC. Among predators, adult Blue Catfish had low MC concentrations, whereas Blue Crabs exhibited high levels of MC in both muscle and viscera. PMID:24694322
Wood, Joseph D; Franklin, Rima B; Garman, Greg; McIninch, Stephen; Porter, Aaron J; Bukaveckas, Paul A
Domoic acid, a neurotoxic amino acid produced by the marine diatom Nitchia pungens multiseries, was determined in samples of anchovies, razor clams, mussels, crab, rat serum, urine and feces by HPLC with UV absorption and electrospray (ESI) mass spectrometric (MS) detection. Shellfish samples were extracted with methanol-water followed by clean-up of the extracts with solid-phase extraction cartridges (strong anion or strong cation exchange). An aliquot of the fraction containing the domoic acid was analysed by HPLC. HPLC column size, mobile phase composition and flow-rate were selected so that essentially the same conditions could be used for both HPLC-UV and HPLC-ESI-MS with selected ion monitoring (SIM) determinations. These included the use of acetonitrile-water-formic acid as the mobile phase, at a flow-rate of 0.2 ml/min (split 13:1 for HPLC-ESI-MS-SIM, 10 microliters/min to the mass spectrometer). The results indicated that extracts found positive by the HPLC-UV method could be readily confirmed directly by HPLC-ESI-MS-SIM without additional sample treatment down to levels of 0.1 micrograms/g of domoic acid. This study demonstrates the use of HPLC-ESI-MS-SIM for the routine confirmation of domoic acid in a wide variety of samples. PMID:8118555
Lawrence, J F; Lau, B P; Cleroux, C; Lewis, D
The Manila clam Ruditapes philippinarum cultivation is an original shellfish farming activity strongly mechanized. In the Chausey archipelago (France) this activity settles on the Lanice conchilega beds, habitat known to host a rich and diversified benthic macrofauna and which is an attractive feeding ground for birds. A first study highlighted that this activity had strong negative effects on the L. conchilega beds and their associated benthic macrofauna. Here we assess the impacts of such an activity on the Eurasian Oystercatcher Haematopus ostralegus for which Chausey is one of the most important national breeding sites and which is also a common species in winter, spring and autumn migrations. We found that Oystercatchers significantly selected the L. conchilega beds to feed and that their spatial distribution was significantly modified after the creation of new clam concessions. In a context of a growing disappearance of pristine coastal ecosystems for the benefit of anthropo-ecosystems, we discuss the problem of the degradation of such benthic habitats with a low resilience which may loose their high functional value. PMID:19100583
Godet, Laurent; Toupoint, Nicolas; Fournier, Jérôme; Le Mao, Patrick; Retière, Christian; Olivier, Frédéric
Mouse bioassay is the official testing method to quantify paralytic shellfish toxins (PSTs) in bivalves. A number of alternative analytical methods have been reported. Some methods have been evaluated by a single laboratory validation. Among the different types of methods, chemical analyses are capable of identifying and quantifying the toxins, however a shortage of the necessary calibration standards hampers implementation of the chemical analyses in routine monitoring of PSTs in bivalves. In our present study, we studied preparation of major PST analogues as calibrants by large-scale cultivation of toxic freshwater cyanobacteria Anabaena circinalis TA04. The cells were steadily grown in 10 L bottle for 28 days. The primary N1-H toxins, C1/C2, were produced at a concentration of 1.3 ±0.1 ?mol/L. The intracellular and extracellular toxins occupied 80% and 20%, respectively. Over 220 ?mol of the toxins was obtained from approximately 200 L of the culture over six months, demonstrating that it is sufficient to prepare saxitoxin analogues. The toxins were chemically converted to six N1-H analogues. Preparation of the analogues was carried out at relatively high yields (50–90%). The results indicate that our preparation method is useful to produce N1-H toxins. In our present study, detailed conditions for preparation of one of the rare N1-H analogues, gonyautoxin-5, were investigated.
Watanabe, Ryuichi; Suzuki, Toshiyuki; Oshima, Yasukatsu
The marine dinoflagellate genus Alexandrium includes a number of species that produce potent neurotoxins responsible for paralytic shellfish poisoning, which in humans may cause muscular paralysis, neurological symptoms and, in extreme cases, death. Because of the genetic diversity of different genera and species, molecular tools may help to detect the presence of target microorganisms in marine field samples. Here we employed a loop-mediated isothermal amplification (LAMP) method for the rapid and simple detection of toxic Alexandrium species. A set of four primers were designed based upon the conserved region of the 5.8S rRNA gene of members of the genus Alexandrium. Using this detection system, toxic Alexandrium genes were amplified and visualized as a ladder-like pattern of bands on agarose gels under isothermal condition within 60 min. The LAMP amplicons were also directly visualized by eye in the reaction tube by the addition of SYBR Green I. This LAMP assay was 10-fold more sensitive than a conventional PCR method with a detection limit of 5 cells per tube when targeting DNA from Alexandrium minutum. The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of target toxic Alexandrium species during coastal water monitoring. PMID:18355290
Wang, Li; Li, Lin; Alam, M J; Geng, Yuhuan; Li, Zhiyong; Yamasaki, Shinji; Shi, Lei
To ensure sustainable uses of the coastal zone, an integrated ecosystemic approach and ecosystem models are required to frame ecological processes and evaluate environmental impacts. Here, a mass-balance trophic (Ecopath) model of the Mont Saint Michel Bay (MSMB) was developed, to analyze the bay's functioning as an ecosystem. This bay, intensively exploited by fishing and for shellfish farming, is also suffering from the proliferation of the gastropod Crepidula fornicata, an exotic species. The MSMB model has 18 compartments, from the primary producers to top predators, and emphasizes the large biomass of filter feeders. The model identified the MSMB as a highly productive ecosystem controlled largely from the bottom-up, and strongly impacted by huge biomasses of filter feeders. However, the low transfer efficiency rates imply that a large part of the primary production is not transferred upward to higher trophic levels, but is lost in high hydrodynamic exchanges and in the trophic impasse represented by a large biomass of Crepidula fornicata.
Arbach Leloup, F.; Desroy, N.; Le Mao, P.; Pauly, D.; Le Pape, O.
GC-ECD methods were adopted to determine the residual level of OCPs (including HCHs and DDTs) and PCBs in the surface sediments collected from mudflat shellfish culturing areas in Ningbo, with the sources of the OCPs and PCBs analyzed and the ecological risks of the residual OCPs and PCBs evaluated. The residual level of OCPs was 0.80-32.40 ng X g(-1), and that of PCBs was 3.20-33.33 ng X g(-1). The HCHs mainly came from long distance atmospheric transportation and historical residues, while the DDTs had new input at some sites, possibly coming from the application of dicofol. At most sites, there existed potential ecological risks of p, p'-DDT and DDTs, with strong indications in Qiangtou and Xidian where the residual level of p, p'-DDT was higher than the effect rang median (ERM), suggesting an ecological menace to the benthos. The residual PCBs at most sites were in low level ecological risk. PMID:22937662
Zhu, Yun-Hai; You, Zhong-Jie; Shentu, Ji-Kang; Zhong, Hui-Ying; Chai, Li-Yue
Laboratory experiments were designed to study the toxin content and profile of the Alexandrium catenella strain ACT03 (isolated from Thau Lagoon, French Mediterranean) in response to abiotic environmental factors under nutrient-replete conditions. This dinoflagellate can produce various paralytic shellfish toxins with concentrations ranging from 2.9 to 50.3 fmol/cell. The toxin profile was characterized by carbamate toxins (GTX3, GTX4 and GTX5) and N-sulfocarbamoyl toxins (C1, C2, C3 and C4). C2 dominated at 12–18 °C, but only for salinities ranging from 10 to 25 psu, whereas GTX5 became dominant at temperatures ranging from 21 to 30 °C at almost all salinities. There was no significant variation in the cellular toxin amount from 18 °C to 27 °C for salinities ranging between 30 and 40 psu. At salinities of 10 to 25 psu, the toxin concentrations always remained below 20 fmol/cell. Toxin content was stable for irradiance ranging from 10 to 70 ?mol photons/m2/s then slightly increased. Overall, the toxin profile was more stable than the toxin content (fmol/cell), except for temperature and/or salinity values different from those recorded during Alexandrium blooms in Thau Lagoon.
Laabir, Mohamed; Collos, Yves; Masseret, Estelle; Grzebyk, Daniel; Abadie, Eric; Savart, Veronique; Sibat, Manoella; Amzil, Zouher
Increased residential development in coastal watersheds has led to increases in anthropogenic nitrogen inputs into estuaries. Sessile bivalves are good candidate organisms to examine animal condition in nutrient-enriched areas because they contribute significantly to energy flow...
Outbreaks of food- and waterborne gastroenteritis are being increasingly reported throughout the world. The analysis of environmental samples by newer diagnostic techniques such as reverse transcription-PCR (RT-PCR) amplification of nucleic acid has begun to identify human enteric viruses (predominantly "Norwalk-like" viruses [NLVs]) as the cause of many of these outbreaks. To streamline NLV detection from environmental samples such as shellfish, we have developed an RT-PCR-oligoprobe amplification and detection method using several new procedures that enable confirmed RT-PCR amplification and product detection in 1 day. The new steps include replacing reverse transcriptase and Taq polymerase with rTth polymerase, a heat-stable enzyme that functions as both a reverse transcriptase and DNA polymerase, in a single-tube, single-buffer, elevated temperature reaction. An internal standard Norwalk virus (NV) RNA control is added to each RT-PCR to identify sample inhibition, and thermolabile uracil N-glycosylase is incorporated into the reaction to prevent PCR product carryover contamination. Finally, RT-PCR-generated amplicons are detected in microtiter wells using virus-specific biotinylated oligoprobes in an enzyme-linked immunosorbent assay-based format. The DNA enzyme immunoassay is based on the capture of PCR product by biotinylated probes fixed onto individual streptavidin-coated wells. Using this method, low levels of NV were detected in stool and both NLV and hepatitis A virus were detected in bivalve mollusks following bioaccumulation. The method also successfully detected NLV in oysters implicated in an outbreak of NLV gastroenteritis. This method dramatically decreases the time needed for analysis and is amenable to automation. PMID:11157239
Schwab, K J; Neill, F H; Le Guyader, F; Estes, M K; Atmar, R L
In recent years, related research has mainly examined lipophilic marine toxins (LMTs) in contaminated bivalves or toxic algae, whereas the levels of LMTs in seawater remain largely unexplored. Okadaic acid (OA), yessotoxin (YTX), and pectenotoxin-2 (PTX2) are three typical LMTs produced by certain marine algae that are closely linked to diarrhetic shellfish poisoning. In this study, a new method of solid phase extraction combined with liquid chromatography - electrospray ionization ion trap tandem mass spectrometry was developed to determine the presence of OA, YTX, and PTX2 in seawater simultaneously. Satisfactory sensitivity, repeatability (RSD<25.00%) and recovery (56.25-70.18%) of the method were achieved. Then, the method was applied to determine the amounts of the three toxins in the coastal seawater. OA and PTX2 were detected in all the seawater samples collected from eight locations along the coastline of Qingdao City, China on October 23, 2012, with concentration ranges of OA 4.24-9.64ngL(-1) and PTX2 0.42-0.74ngL(-1). Monthly concentrations of OA and PTX2 in the seawater of four locations were determined over the course of a year, with concentration ranges of OA 1.41-89.52ngL(-1) and PTX2 below detectable limit to 1.70ngL(-1). The peak values of OA and PTX2 in coastal seawater were observed in August and July, respectively. Our results suggest that follow-up research on the fate modeling and risk assessment of LMTs in coastal seawater should be implemented. PMID:24997966
Li, Xin; Li, Zhaoyong; Chen, Junhui; Shi, Qian; Zhang, Rutan; Wang, Shuai; Wang, Xiaoru
Concentrations of paralytic shellfish toxins (C1+2, B1, dcGTX2+3, dcSTX, GTX2+3 and STX) were determined by LC-FLD in composite samples of digestive glands of the cockle Cerastoderma edule and in each sub-cellular particulate fractions obtained after differential centrifugation (nuclei+debris, mitochondria, lysosomes and microsomes). The specimens were sampled during the exposure to a bloom of Gymnodinium catenatum (day 0) and in the subsequent 8, 12, 14, 19, 21 and 25 days under natural depuration conditions. Toxin profiles of digestive glands were dominated by C1+2 followed by B1 and dcGTX2+3, although the proportion between C1+2 and B1 contents decreased with the time, indicating a slower elimination of B1. All toxins, except GTX2+3 and STX, were quantified in the four sub-cellular fractions. The content of the quantified toxins decreased most markedly in nuclei+debris and microsomal fractions, during the first eight and 12 days, respectively. Conversely, different patterns were observed among toxins in mitochondrial and lysosomal fractions. The less accentuated decreases of dcGTX2+3 and dcSTX contents in the mitochondrial fraction may have resulted from the conversion of other toxins, like C1+2 and B1, associated with enzymatic activities in that fraction. The largest discrepancy was registered in lysosomal fraction for B1, since its content increased after eight days of post-bloom conditions. Input of B1 may come from the conversion of other toxins, like the abundant B2 and C1+2. These transformations are associated to the major role of lysosomes in the intra-cellular digestive process of materials acquired through vesicular transport. PMID:24736027
Botelho, Maria João; Raimundo, Joana; Vale, Carlos; Ferreira, João Gomes
The biotoxins, azaspiracids (AZAs), from marine phytoplankton accumulate in shellfish and affect human health by causing severe gastrointestinal disturbance, diarrhea, nausea and vomiting. Specific and sensitive methods have been developed and validated for the determination of the most commonly occurring azaspiracid analogs. An LTQ Orbitrap mass spectrometer is a hybrid instrument that combines linear ion trap (LIT) mass spectrometry (MS) with high-resolution Fourier transform (FT) MS and this was exploited to perform simultaneous ultra-high-resolution full-scan MS analysis and collision-induced dissociation (CID) tandem mass spectrometry (MS/MS). Using the highest mass resolution setting (100,000 FWHM) in full-scan mode, the methodology was validated for the determination of six AZAs in mussel (Mytilus galloprovincialis) tissue extracts. Ultra-high mass resolution, together with a narrow mass tolerance window of ±2 mDa, dramatically improved detection sensitivity. In addition to employing chromatographic resolution to distinguish between the isomeric azaspiracid analogs, AZA1/AZA6 and AZA4/AZA5, higher energy collisionally induced dissociation (HCD) fragmentation on selected precursor ions were performed in parallel with full-scan FTMS. Using HCD MS/MS, most precursor and product ion masses were determined within 1?ppm of the theoretical m/z values throughout the mass spectral range and this enhanced the reliability of analyte identity.For the analysis of mussels (M. galloprovincialis), the method limit of quantitation (LOQ) was 0.010?µg/g using full-scan FTMS and this was comparable with the LOQ (0.007?µg/g) using CID MS/MS. The repeatability data were; intra-day RSD% (1.8-4.4%; n?=?6) and inter-day RSD% (4.7-8.6%; n?=?3). Application of these methods to the analysis of mussels (M. edulis) that were naturally contaminated with azaspiracids, using high-resolution full-scan Orbitrap MS and low-resolution CID MS/MS, produced equivalent quantitative data. PMID:20872629
Skrabáková, Zuzana; O'Halloran, John; van Pelt, Frank N A M; James, Kevin J
To protect public health from the potential risk of paralytic shellfish poisoning (PSP) in sea scallops, Placopecten magellanicus, from the Atlantic offshore U.S. waters of Georges Bank, harvesting of roe-on or whole scallops is banned. Only adductor muscles may be sold if harvested from Georges Bank, Gulf of Maine, or the PSP closure areas as far west as 71° West Longitude. Given the limited toxicity data available for sea scallops from this region both prior to and subsequent to implementation of this management strategy, this study sought a more extensive spatial and temporal evaluation of sea scallop gonad and viscera toxicities that would inform management decisions related to the roe-on and whole scallop fishery. Both overall toxicity and toxin composition were measured for sea scallop gonads and viscera collected from 232 stations in 2007 and 23 in 2010. Overall toxicity was assessed using two screening methods: field-deployable Jellett Rapid Tests (JRT) and quantitative, laboratory-based receptor binding assays (in 2007). Additionally, a quantitative liquid chromatography with fluorescence detection (LC-FD) method was used to determine toxin composition and overall toxicity (in 2010). The at-sea qualitative JRT screening tool results, whereby a positive indicated the sample contained at least 40 ?g saxitoxin (STX) equivalents 100 g-1, were often inconsistent with results obtained using the quantitative methods. Sea scallop viscera toxicity represented the majority of toxin load in the organism and was often in excess of the regulatory guidance level. Sea scallop gonads accounted for a small percentage of total toxicity, but at times reached unsafe levels. Toxin composition in both the gonads and viscera was dominated by STX and gonyautoxin 2/3, as has been reported in previous studies. No predictive indices of gonad toxicity were found. Results at this time do not support a roe-on or whole scallop fishery on Georges Bank. While access restrictions to whole scallops on Georges Bank are unlikely to change based on these results, it seems plausible that a management strategy requiring at-sea testing of scallop gonads for PSP may be necessary in order for a safe roe-on scallop fishery from this region to be feasible.
DeGrasse, Stacey; Vanegas, Camilo; Conrad, Stephen
Farmers' vigilance is essential for the detection of epidemics, including potential emerging diseases, in marine shellfish. A field study was conducted to investigate oyster farmers' reporting practices and behaviour, and to identify factors influencing the reporting process of oyster mortality, with the ultimate aim of improving early detection of unexplained oyster mortality outbreaks. A retrospective case-control study of oyster farmers from Charente-Maritime (France) was designed, based on interviews with 27 non-reporting and 89 reporting farmers, further split into 40 formerly-reporting and 49 currently-reporting farmers. Information about farmer and farm characteristics, farming practices, farm health history and related financial compensation on the farm, knowledge of the mortality reporting system and reporting behaviour was collected. Sampling design was considered in the calculations and farmers' reporting behaviour was modelled using an ordinal logistic regression (continuation-ratio model). Notification procedures were fairly well known among farmers and the reporting system was well accepted overall. Nevertheless, a lack of awareness of the aims of the reporting system was revealed, which contributed to late reporting. Factors identified as driving a farmer's decision to report oyster mortality concerned their lack of awareness of mortality reporting (production type, farm size, location of the production cycle, accessibility of the leasing grounds) and willingness to report (possibility and extent of financial compensation, a feeling of not being involved, whether it was first year of reporting). Overall classification performance of the model built in this study was 64%. In particular, financial compensation for oyster production losses appeared to be a clear incentive for reporting, but was countered by a habituation effect combined with a lack of awareness of the aims of the reporting system: oyster farmers looking for benefits for themselves in reporting, rather than early detection of a disease outbreak. Both economic compensation and the farmers' non-economic values and perceptions should be considered to improve oyster farmers' reporting compliance and sustainability of the reporting system. Education and participatory approaches could help to change these attitudes and thus improve oyster farmers' compliance with reporting duties, resulting in improved early detection of epidemics and emerging or exotic oyster diseases. PMID:24880624
Lupo, C; Osta Amigo, A; Mandard, Y V; Peroz, C; Renault, T
The objective of this study was to identify and characterize a "biomarker of toxicity" from the proteome of Alexandrium tamarense, a paralytic shellfish toxin (PST)-producing dinoflagellate. A combination of 2-DE and MS approaches was employed to identify proteins of interest in the vegetative cells of several strains of A. tamarense with different toxin compositions and from different geographical locations. The electrophoretic analysis of the total water-soluble proteins from these toxic strains by 2-DE showed that several abundant proteins, namely AT-T1, AT-T2 and AT-T3, differing slightly in apparent Mr and pIs, were consistently present in all toxic strains of A. tamarense. Further analysis by MALDI-TOF MS and N-terminal amino acid sequencing revealed that they are isoforms of the same protein. Even more intriguing is that these proteins in A. tamarense have similar amino acid sequences and are closely related to a "biomarker of toxicity" previously reported in A. minutum. Unambiguous and highly species-specific identification was later achieved by comparing the PMFs of proteins in these two species. An initial attempt to characterize these proteins by generation of murine polyclonal antibodies against the AT-T1 protein was successful. Western blot analysis using the murine AT-T1-polycolonal antibodies identified all the toxic strains of A. tamarense and A. minutum, but not the nontoxic strain of A. tamarense. These results indicate that these protein characteristics for toxic strains are species-specific and that they are stable properties of the tested algae which are clearly distinguishable irrespective of geographical location and toxin composition. To our knowledge, this is the first study to demonstrate the use of polyclonal antibodies against marker proteins purified from 2-DE gels to distinguish different strains and species of the PST-producing dinoflagellate Alexandrium. It provides the basis for the production of monoclonal antibody probes against the "biomarkers of toxicity" for those dinoflagellates whose genome is incompletely characterized. Potentially, immunoassays could be developed to detect the presence of toxic algae in routine monitoring programs as well as to predict bloom development and movement. PMID:16342137
Chan, Leo Lai; Sit, Wai-Hung; Lam, Paul Kwan-Sing; Hsieh, Dennis Paul Hsientang; Hodgkiss, Ivor John; Wan, Jennifer Man-Fan; Ho, Alvin Yam-Tat; Choi, Nicola Man-Chi; Wang, Da-Zhi; Dudgeon, David
This State of New Jersey fact sheet (PDF) briefly outlines the ecological and socioeconomic risks associated with Quahog Parasite X (QPX), a parasite that kills hard clams. QPX was first discovered in New Jersey in the 1970s.
Jersey, State O.
The invasive Asian shore crab, Hemigrapsus sanguineus, has recently been observed occupying salt marshes, a novel environment for this crab species. As it invades this new habitat, it is likely to interact with a number of important salt marsh species. To understand the potential effects of H. sanguineus on this ecosystem, interactions between this invasive crab and important salt marsh ecosystem engineers were examined. Laboratory experiments demonstrated competition for burrows between H. sanguineus and the native fiddler crab, Uca pugilator. Results indicate that H. sanguineus is able to displace an established fiddler crab from its burrow. Feeding experiments revealed that the presence of H. sanguineus has a significantly negative impact on the number as well as the biomass of ribbed mussels (Geukensia demissa) consumed by the green crab, Carcinus maenas, although this only occurred at high predator densities. In addition, when both crabs foraged together, there was a significant shift in the size of mussels consumed. These interactions suggests that H. sanguineus may have long-term impacts and wide-ranging negative effects on the saltmarsh ecosystem.
Fournier, Alexa M.; Furman, Bradley T.; Carroll, John M.
Byssally tethered mussels are found in a variety of habitats, including rocky intertidal, salt marsh, subtidal, and hydrothermal vents. One key to the survival of mussels in these communities is a secure attachment, achieved by the production of byssal threads. Although many studies have detailed the unique biomechanical properties of byssal threads, only a few prevalent species have been examined. This study assesses the variation in the mechanical properties of byssus in a broad range of mussel species from diverse environments, including intertidal and subtidal Mytilus edulis, Modiolus modiolus, Geukensia demissa, Bathymodiolus thermophilus, and Dreissena polymorpha. A tensometer was used to measure quasi-static and dynamic mechanical properties of individual threads, and several aspects of morphology were quantified. The results indicate that thread mechanical properties vary among mussel species, and several novel properties were observed. For example, of the species examined, D. polymorpha threads were the strongest, stiffest, least resilient, and fastest to recover after partial deformation. Threads of M. modiolus were characterized by the presence of two distinct yield regions prior to tensile failure. This comparative study not only provides insight into the ecological limitations and evolution of mussels, but also suggests new models for the design of novel biomimetic polymers. PMID:17179385
Brazee, Shanna L; Carrington, Emily
New Bedford Harbor (NBH), MA, is a Superfund site because of high polychlorinated biphenyl (PCB) concentrations in the sediment. From April 1994 to September 1995, a remedial dredging operation (termed the "Hot Spot") removed the most contaminated sediments (PCB concentrations greater than 4000 microg/g) from the upper harbor. During remediation, a monitoring program assessed the potential environmental impacts to NBH and adjacent Buzzards Bay. The monitoring program was developed with input from federal, state, and local authorities. Site-specific decision criteria were established to assess net PCB transport, water column toxicity, and PCB bioaccumulation in blue and ribbed mussels (Mytilus edulis and Geukensia demissa, respectively). The remediation was completed without exceeding PCB net transport or acute toxicity effects specified in the decision criteria. In addition, PCB bioaccumulation in mussels during this time period was not significantly greater than pre- or post-operational measurements. The results indicated that approximately 14000 cubic yards of highly PCB contaminated sediment were permanently removed with minimal environmental effects. The lessons learned during this operation, as well as previous pilot studies at the site, will be used to make full-scale remedial efforts in NBH more efficient and environmentally protective. PMID:16311831
Bergen, Barbara J; Nelson, William G; Mackay, Joseph; Dickerson, David; Jayaraman, Saro
Detection of okadaic acid and related esters in mussels during diarrhetic shellfish poisoning (DSP) episodes in Greece using the mouse bioassay, the PP2A inhibition assay and HPLC with fluorimetric detection.
An approach involving chemical, functional and biological techniques was taken for the detection and quantification of the marine toxin okadaic acid (OA) in mussels from Thermaikos and Saronikos Gulfs, Greece, during DSP episodes that occurred in 2006-2007. Samples were analyzed using the mouse bioassay, high performance liquid chromatography with fluorimetric detection (HPLC-FLD), using l-bromoacetylpyrene (BAP), as a precolumn derivatisation reagent, and the protein phosphatase 2A inhibition assay (PP2AIA) using a commercially available kit. Okadaic acid (OA) and its polar and non-polar esters were detected and quantified by HPLC-FLD, after hydrolysis of the samples during preparation. The detection limit of the HPLC method for OA was 5.86 microg OA/kg, which permits this method to be used for the regulatory control of these toxins in shellfish. Comparison of the results by all three methods revealed excellent consistency. PMID:19046982
Prassopoulou, Eleanna; Katikou, Panagiota; Georgantelis, Dimitrios; Kyritsakis, Apostolos
An integrated multi-trophic aquaculture assessment for Pacific oyster (Crassostrea gigas) aquaculture as a bioremediation strategy in areas impacted by fish farm effluents in Rio San Pedro was assessed by combining geographic information system with carrying capacity models. Sites of 0.44 km(2) were evaluated considering constraints; physical factors, growth and survival factors, environmental quality factors, water and sediment quality criteria, factor suitability ranges, and Multi-Criteria Evaluation. Isleta and Flamenco are promising sites for oyster production, and Dorada is of marginal interest. Carbon and nitrogen removal from the water by algae and through detritus filtration was estimated. The biodeposition of organic material from longline leases was found to have little negative impact on sediment. The eutrophication results indicate that phytoplankton removal had a positive impact on water quality at the Dorada. This case study quantified the direct profitability and bioremediative environmental service advantages that fish-shellfish farms can have relative to fish monocultures. PMID:22310375
Silva, C; Yáñez, E; Martín-Díaz, M L; DelValls, T A
Refinement of AOAC Official Method 2005.06 liquid chromatography-fluorescence detection method to improve performance characteristics for the determination of paralytic shellfish toxins in king and queen scallops.
AOAC Official Method 2005.06 LC-fluorescence detection (FLD) method is an official alternative to the mouse bioassay for the determination of paralytic shellfish poisoning (PSP) toxins in bivalve shellfish. To validate the method for species of relevance to the UK official control monitoring program, the method performance characteristics were tested for whole king and queen scallops. Validation showed that, while the performance was generally acceptable for the quantitation of non-N-hydroxylated toxins, poor toxin recovery and sensitivity was evident for the analysis of N-hydroxylated toxins following periodate oxidation. These effects occurred in a range of scallop samples with variable temporal and spatial sources. The effects were also noted in other laboratories following a small interlaboratory study. As a result, the method was refined to improve the recovery and sensitivity of analysis following the periodate oxidation step in the PSP method for scallops. Performance improved through alterations to the preparation of the periodate oxidant, use of higher volumes for C18 cleanup, and injection volumes in combination with the use of a king scallop matrix modifier for oxidation of N-hydroxylated toxin calibration standards. A single-laboratory validation of the refined method showed that the selectivity, linearity, sensitivity, recovery, and precision were acceptable and similar to values reported previously for AOAC Official Method 2005.06 in other bivalve species. Results showed the method to be rugged for all parameters investigated, including small changes to the composition of the new periodate reagent utilized in the refined method. The refined scallops LC method was subsequently compared with the European reference method. PSP-positive scallops showed an excellent agreement between the methods for queen and Atlantic scallops, with a small level of positive bias in the LC results for whole king scallops. These differences were related solely to the use of the highest toxicity equivalence factors for toxin epimeric pairs, with gonyautoxin (GTX)1,4 and GTX2,3 in particular present at high concentrations in the king scallops. Overall, the refined LC-FLD method improved the performance characteristics of AOAC Official Method 2005.06 for the determination of PSP toxins in whole king and queen scallops, and showed a good overall agreement between the official methodologies. It is, therefore, recommended as a more appropriate option for the routine monitoring of PSP toxins in these species. PMID:22468351
Turner, Andrew D; Hatfield, Robert G
Database of radionuclide measurements in Columbia River water, fish, waterfowl, gamebirds, and shellfish downstream of Hanford`s single-pass production reactors, 1960--1970. Hanford Environmental Dose Reconstruction Project
This report is a result of the Hanford Environmental Dose Reconstruction (HEDR) Project. The goal of the HEDR Project is to estimate the radiation dose that individuals could have received from radionuclide emissions since 1944 at the Hanford Site near Richland, Washington. The HEDR Project is conducted by Battelle, Pacific Northwest Laboratories. The time periods of greatest interest to the HEDR study vary depending on the type of environmental media concerned. Concentrations of radionuclides in Columbia River media from 1960--1970 provide the best historical data for validation of the Columbia River pathway computer models. This report provides the historical radionuclide measurements in Columbia River water (1960--1970), fish (1960--1967), waterfowl (1960--1970), gamebirds (1967--1970), and shellfish (1960--1970). Because of the large size of the databases (845 pages), this report is being published on diskette. A diskette of this report is available from the Technical Steering Panel (c/o K. CharLee, Office of Nuclear Waste Management, Department of Ecology, Technical Support and Publication Information Section, P.O. Box 47651, Olympia, Washington 98504-7651).
Thiede, M.E.; Duncan, J.P.
Predators can affect prey populations and, via trophic cascades, predators can indirectly impact resource populations (2 trophic levels below the predator) through consumption of prey (density-mediated indirect effects; DMIEs) and by inducing predator-avoidance behavior in prey (trait-mediated indirect effects; TMIEs). Prey often employ multiple predator-avoidance behaviors, such as dispersal or reduced foraging activity, but estimates of TMIEs are usually on individual behaviors. We assessed direct and indirect predator effects in a mesocosm experiment using a marine food chain consisting of a predator (toadfish – Opsanus tau), prey (mud crab - Panopeus herbstii) and resource (ribbed mussel – Geukensia demissa). We measured dispersal and foraging activity of prey separately by manipulating both the presence and absence of the predator, and whether prey could or could not disperse into a predator-free area. Consumption of prey was 9 times greater when prey could not disperse, probably because mesocosm boundaries increased predator capture success. Although predator presence did not significantly affect the number of crabs that emigrated, the presence of a predator decreased resource consumption by prey, which resulted in fewer resources consumed for each prey that emigrated in the presence of a predator, and reduced the overall TMIE. When prey were unable to disperse, TMIEs on mussel survival were 3 times higher than the DMIEs. When prey were allowed to disperse, the TMIEs on resource survival increased to 11-times the DMIEs. We found that restricting the ability of prey to disperse, or focusing on only one predator-avoidance behavior, may be underestimating TMIEs. Our results indicate that the relative contribution of behavior and consumption in food chain dynamics will depend on which predator-avoidance behaviors are allowed to occur and measured.
Geraldi, Nathan R.; Macreadie, Peter I.
Activity measurements of a suite of radionuclides (241Am, 239,240Pu, 238Pu, 238U, 234U, 235U, 232Th, 230Th, 228Th, 228Ra, 137Cs, 210Pb, 90Sr and 40K) in biota reference material (Ocean Shellfish): CCRI(II)-S3
In 2005, the CCRI decided that a comparison undertaken from 2002 to 2008 by the NIST (under the auspices of the Inter-America Metrology System [SIM]) in the development of a new biota (Ocean Shellfish) standard reference material (SRM) was sufficiently well constructed that it could be converted into a supplementary comparison under CCRI(II), with comparison identifier CCRI(II)-S3. This would enable the comparison to be used to support calibration and measurement capability (CMC) claims for radionuclide measurements in reference materials (specifically, animal-based organic materials). Previous comparisons of radionuclides have been of single or multiple nuclides in non-complex matrices and results of such could not be extended to support capabilities to measure the same nuclides in reference materials. The results of this comparison have been used to determine the certified reference value of the SRM. The key comparison working group (KCWG) of the CCRI(II) has approved this approach as a mechanism to link all the results to certified 'reference values' in lieu of the key comparison reference value (KCRV) of these specified radionuclides in this type of matrix (shellfish) so as to support CMCs of similar materials submitted by the present participants. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).
Nour, S.; Karam, L. R.; Inn, K. G. W.
Habitat provides a variety of life support functions for many species, such as providing shelter, substrate, food, and nursery areas. Habitat alteration is one of the most important causes of declines in ecological resources in North America, and habitats essential to the well b...
The use of remote sensing techniques for collecting bacteriological, physical, and chemical water quality data, locating point and nonpoint sources of pollution, and developing hydrological data was found to be valuable to the Maryland program if it could be produced effectively and rapidly with a minimum amount of ground corroboration.
Approximately 50 ka, one or more subgroups of modern humans expanded from Africa to populate the rest of the world. Significant behavioral change accompanied this expansion, and archaeologists commonly seek its roots in the African Middle Stone Age (MSA; ?200 to ?50 ka). Easily recognizable art objects and “jewelry” become common only in sites that postdate the MSA in Africa and Eurasia, but some MSA sites contain possible precursors, especially including abstractly incised fragments of ocher and perforated shells interpreted as beads. These proposed art objects have convinced most specialists that MSA people were behaviorally (cognitively) modern, and many argue that population growth explains the appearance of art in the MSA and its post-MSA florescence. The average size of rocky intertidal gastropod species in MSA and later coastal middens allows a test of this idea, because smaller size implies more intense collection, and more intense collection is most readily attributed to growth in the number of human collectors. Here we demonstrate that economically important Cape turban shells and limpets from MSA layers along the south and west coasts of South Africa are consistently and significantly larger than turban shells and limpets in succeeding Later Stone Age (LSA) layers that formed under equivalent environmental conditions. We conclude that whatever cognitive capacity precocious MSA artifacts imply, it was not associated with human population growth. MSA populations remained consistently small by LSA standards, and a substantial increase in population size is obvious only near the MSA/LSA transition, when it is dramatically reflected in the Out-of-Africa expansion.
Klein, Richard G.; Steele, Teresa E.
Preterm parturition is a syndrome caused by several mechanisms of disease, including intrauterine infection/inflammation, uteroplacental ischemia, uterine overdistension, cervical disease, maternal/fetal stress, abnormal allogeneic responses, allergic reactions, and unknown insults. An allergic-like mechanism was proposed as a potential etiology for the preterm parturition syndrome, based on the observation that eosinophils were present in the amniotic fluid in a fraction of women with preterm labor and a history of allergy, coupled with the observation that conditioned media from degranulated mast cells (the effector cells of type 1 hypersensitivity) induced contractility of human myometrial strips. This communication describes a case of a pregnant woman who had an allergic reaction and regular uterine contractions after the ingestion of lobster meat, to which she was known to be allergic. Preterm labor subsided after the treatment of antihistamines and steroids. The patient subsequently delivered at term. At follow-up, the child was diagnosed with atopy and asthma, and required frequent use of inhaled corticosteroids and beta-2 adrenergic agents.
ROMERO, ROBERTO; KUSANOVIC, JUAN PEDRO; MUNOZ, HERNAN; GOMEZ, RICARDO; LAMONT, RONALD F.; YEO, LAMI
Aquaculture, the farming of aquatic organisms, is more prevalent in the rest of the world, particularly in East Asia, than in the United States. Aquaculture began to receive focused attention in the United States during the 1970s when many domestic fisher...
Contents: Membrane filter technics as applied to estuarine and tributary waters; Bacteriological studies on commercial processing of Olympia oysters (Ostrea lurida); Storage studies on shucked olympia oysters (Ostrea lurida); Storage studies on olympia oy...
W. J. Beck J. C. Hoff
Contents: Comparative study of several indicator organisms in samples collected from three northwest watersheds; Studies on the incidence of Clostridium botulinum Type E in some west coast estuarine areas; Studies on wet storage of oyster and clam shellst...
W. J. Beck J. C. Hoff
The Bartol in-air Pixe system has been used to study the elemental composition of many varieties of sea life; the sensitivity for the elements from Na to Sr is good, a reasonable spatial resolution is attainable and little preparation of the samples is required. Two examples are cited. First, the composition of the inner nacreous and prismatic regions of mussels ( Mytilus edulis) taken monthly (1981-1982) from the Damariscotta River, Maine, has been measured and, second, the shell and soft tissues of oysters ( Crassostrea virginica) grown under laboratory conditions on substrate of compacted residues from coal-fired power plants have been analysed. The mussel study shows a rather large anomaly in the Sr/Ca ratio for the nacreous region for May which suggests a possible connection with the reproduction cycle. Oysters grown on metal-containing coal-ash substrata gave tissue and shell metal concentrations similar to natural oysters, an indication of no anomalous uptake of metals from the coal-ash substrata.
Swann, C. P.; Hansen, K. Mueller; Price, K.; Lutz, R.
A tertiary treatment plant and an ultraviolet disinfection chamber were installed following an activated sludge plant at the municipal sewage treatment plant in St. Michaels, Maryland. The multiple-tube fermentation technique was used to determine the tot...
J. A. Roeber F. M. Hoot
The following case concerns a soft tissue Vibrio cholerae (V. cholerae) infection in a fisherman who cut his foot while retrieving his fishing dinghy. It is rare for V. cholerae to cause extraintestinal infection. This V. cholera was identified as a non-toxigenic organism. The patient was successfully treated with medical therapy at Waikato Hospital (Hamilton, New Zealand) and discharged home after 10 days. PMID:24045356
Whittaker, Samuel J
It has been demonstrated that several microalgae species, which are thought to be good food for feeders, will survive and grow in Roswell Test Facility (RTF) saline ground water at pilot scale (50 sq m raceways). Under specific environmental and cultural ...
Candida parapsilosis, C. tropicalis, and Torulopsis glabrata were the human-associated yeasts most frequently isolated from quahogs, oysters, and mussels collected from four estuarine areas along the northern shore of Long Island Sound. Some inconsistency and seasonal variation in the occurrence of these and other yeast species were noted. In particular, C. albicans densities were greatest during colder months in the more heavily polluted waters. A total of 347 yeasts were isolated and cultured at 37 degrees C and, of these, 219 of 62% were human-associated forms. Generally, these yeasts in the animals sampled reflected the overall pollution status of the estuary from which they were taken. This study represents a clear demonstration of potentially pathogenic yeasts in a valuable marine resource.
Buck, J D; Bubucis, P M; Combs, T J
Maximum acceptable concentrations of metals in food - based on total concentrations - have been established in many countries. To improve risk assessment, it would be better to take into account bioaccessible concentrations. A total of seven species of molluscs from France, UK and Hong Kong was examined in this study including clams, mussels, oysters, scallops and gastropods. The species which have total metal concentrations higher than the most severe food security limits are mainly oysters (all of the three studied species), the gastropod Buccinum undatum for cadmium and zinc, and scallops for cadmium. The lowest bioaccessibility (in % extractability with gut juices) was observed for silver (median for all of the species: 14%), it was moderate for lead (median: 33%) and higher for cadmium, zinc and copper (medians were respectively 54%, 65%, and 70%). In most cases, bioaccessible concentrations remained higher than the safety limits, except for cadmium in scallops and zinc in B. undatum. The influence of feeding habit (masticated or swallowed, addition of vinegar or lemon) on metal bioaccessibility in oysters is limited. On the contrary, cooking the gastropods decreased the bioaccessibility of metals, except silver. PMID:18329777
Amiard, Jean-Claude; Amiard-Triquet, Claude; Charbonnier, Laetitia; Mesnil, Aurélie; Rainbow, Philip S; Wang, Wen-Xiong
ECD?GC, NP?FID?GC and GC\\/MS assay which make it possible to concurrently determine oxadiazon and its metabolites (phenol, mehoxy, acid and alcohol forms) in pond snails (collected in the Yodo River) and blue mussels (collected in the Osaka Bay) were developed. These assays were used to determine oxadiazon and its four metabolites in pond snails and blue mussels collected during the
Y. Murakami; T. Nishimune; K. Sueki
This work had as main objectives to characterize two bacteriocins produced by lactic acid bacteria (LAB) previously isolated from non-fermented seafood, in order to evaluate their potential as new food protective agents. The two bacteriocinogenic isolates were identified by Polymerase Chain Reaction (PCR) using genus- and species-specific primers, and confirmed by 16S rDNA sequencing, as Enterococcus faecium and Pediococcus pentosaceus.
Ana Luísa Pinto; Melissa Fernandes; Cristina Pinto; Helena Albano; Fernanda Castilho; Paula Teixeira; Paul A. Gibbs
The project will document the adverse effects of episodic hypoxia on the biotic integrity of Lake Pontchartrain and provide quantitative data on environmental benefits derived from the restoration of 100 square miles of clam habitat in Lake Pontchartrain. This project will prov...
The blooms of toxic cyanobacteria (blue–green algae) are causing problems in many countries. During a screening of toxic freshwater cyanobacteria in Brazil, three strains isolated from the State of São Paulo were found toxic by the mouse bioassay. They all were identified as Cylindrospermopsis raciborskii by a close morphological examination. Extracts of cultured cells caused acute death to mice when
Néstor Lagos; Hideyuki Onodera; Pedro Antonio Zagatto; Dar??o Andrinolo; Sandra M. F. Q Azevedo; Yasukatsu Oshima
Studies by the University of Lowell Radiation Laboratory and the US National Marine Fisheries Service N.E. Laboratory in Gloucester, MA on softshelled clams (Mya arenaria) demonstrated the effectiveness of low to medium doses of Cobalt 60 source gamma irr...
L. E. Beghian J. C. Mallett
Bandon Bay (Surat Thani Province) is one of the most productive coastal areas in southern Thailand. The Tapi River and 18\\u000a channels are the main sources of freshwater, nutrients, organic matter and sediment to the bay and the loading of freshwater\\u000a and nutrients provide essential support for the production of phytoplankton in the estuarine ecosystem. Bandon Bay is important\\u000a as
A. Jarernpornnipat; O. Pedersen; K. R. Jensen; S. Boromthanarat; S. Vongvisessomjai; P. Choncheanchob
... the harmful effects of mercury. ?? Do not eat: • Shark • Swordfish • King Mackerel • Tilefish They contain high levels ... time to accumulate it. These large fish (swordfish, shark, king mackerel and tilefish) pose the greatest risk. ...
Conditions for the determination of lipophilic marine toxins, such as yessotoxins and pectenotoxins (PTX)-6, were investigated with capillary electrophoresis coupled to mass spectrometry (MS) with an electrospray ionization source. After optimization, a simple and MS compatible alkaline volatile buffer solution of ammonium acetate was selected as background electrolyte, with isopropanol\\/water (80\\/20, v\\/v) sheath liquid modified with ammonium acetate used at
P. de la Iglesia; A. Gago-Martínez
Levels of 18 polychlorinated biphenyl (PCB) congeners were determined by gas chromatography-mass spectrometry (GC-MS) in some marine species, living both in the coastal area and in deeper seawater. In some species analysis was performed separately in edible parts (fillets) and in viscera. The existence and degree of bioaccumulation was assessed studying individual species of very different size, with the smaller being younger. Furthermore, with a multivariate statistical analysis, a correlation between PCB congeners and the feeding habits and habitat of the fish was demonstrated. The results show that fat from edible parts (fish fillets) had total PCB levels in the range 22.6-601.9 µg kg(-1) (with 601.9 µg kg(-1) in anchovies), while fat from viscera showed much higher concentrations (407.3-916.6 µg kg(-1)). Bioaccumulation was confirmed, comparing PCB levels between younger and older individual hake, squid, and horned octopus. The total PCB concentration ratio (older/younger individuals) ranges from 2.11 (squid = 292.1/137.8 µg kg(-1)) to 3.46 (hake = 546.0/158.0 µg kg(-1)). PMID:24784539
Sagratini, G; Buccioni, M; Ciccarelli, C; Conti, P; Cristalli, G; Giardinà, D; Lambertucci, C; Marucci, G; Volpini, R; Vittori, S
The impact of suspended mussel culture (Mytilus edulis, M. trossulus) on the benthos of a small Nova Scotia cove (7 m depth) was assessed using meehods involving both benthic metabolism and\\u000a community structure. Due to deposition of mussel feces and pseudofeces, sedimentation rate was higher under the mussel culture\\u000a lines than at an adjacent reference site of similar sediment texture.
J. Grant; A. Hatcher; D. B. Scott; P. Pocklington; C. T. Schafer; G. V. Winters
Temperature regulation is a vital component of any aquaculture system. Existing facilities can be retrofitted with extra insulation, waste heat recovery systems and in some cases, active solar water heating. Those aquaculture ventures that seek to raise o...
D. L. Berg
To supplement natural hard clam (Mercenaria mercenaria) production, Brookhaven Township constructed an onland facility to culture seed clasm in 1980. The facility is only operated from June to October when water temperatures are above 60/sup 0/F, the minimum acceptable for hard clam growth. If seawater could be heated economically, operation of the facility could begin in March, thereby increasing hard clam production. The use of solar energy is particularly suited for this purpose since the seawater need only be warmed a few degrees. After considering a number of active and passive type systems, the Town opted for the latter since it would be simpler to build and operate, less expensive, and at the same time would meet the biological requirements of the hard clam. The two basic components of the solar pond, as the system is known, are a reservoir in which the seawater is heated by direct absorption and a transparent cover to minimize heat loss. For an initial production of 1 million .75mm seed clams, two tanks each measuring 10 feet by 20 feet by 3 feet deep were installed beneath a greenhouse type structure. Total cost of the system was less than $23,000.
...an antler, the length of which is greater than its width and is at least...the skin of the head (cape) or the entire...skin, viscera, head, or bones of...hares, marmots, beaver, muskrats, or...as well as the head, tail, fins, and...
This Woods Hole Oceanographic Institute news release reports that a genetic test has been developed to detect QPX (Quahog Parasite Unknown), a crippling disease occurring in clam beds from Cape Cod south to Virginia and north to Canada. The article features Woods Hole scientists Rebecca Gast and Roxanna Smolowitz and links to the related Oceanus magazine article.
Dawicki, Shelley; Institute, Woods H.
...norovirus-surv-disease-burden.htm. 3. National Microbiology Laboratory and Public Health Agency...Estuary,'' Applied and Environmental Microbiology, vol. 69, pp. 7130-7136, 2003...Illness,'' Applied Environmental Microbiology, vol. 66, pp. 1375-1378,...
Ocean surface pH levels are predicted to fall by 0.3–0.4 pH units by the end of the century and are likely to coincide with an increase in sea surface temperature of 2–4°C. The combined effect of ocean acidification and warming on the functional properties of bivalve shells is largely unknown and of growing concern as the shell provides protection from mechanical and environmental challenges. We examined the effects of near-future pH (ambient pH –0.4 pH units) and warming (ambient temperature +4°C) on the shells of the commercially important bivalve, Mytilus edulis when fed for a limited period (4–6 h day?1). After six months exposure, warming, but not acidification, significantly reduced shell strength determined as reductions in the maximum load endured by the shells. However, acidification resulted in a reduction in shell flex before failure. Reductions in shell strength with warming could not be explained by alterations in morphology, or shell composition but were accompanied by reductions in shell surface area, and by a fall in whole-body condition index. It appears that warming has an indirect effect on shell strength by re-allocating energy from shell formation to support temperature-related increases in maintenance costs, especially as food supply was limited and the mussels were probably relying on internal energy reserves. The maintenance of shell strength despite seawater acidification suggests that biomineralisation processes are unaffected by the associated changes in CaCO3 saturation levels. We conclude that under near-future climate change conditions, ocean warming will pose a greater risk to shell integrity in M. edulis than ocean acidification when food availability is limited.
Mackenzie, Clara L.; Ormondroyd, Graham A.; Curling, Simon F.; Ball, Richard J.; Whiteley, Nia M.; Malham, Shelagh K.
The worldwide depletion of major fish stocks through intensive industrial fishing is thought to have profoundly altered the trophic structure of marine ecosystems. Here we assess changes in the trophic structure of the English Channel marine ecosystem using a 90-year time-series (1920–2010) of commercial fishery landings. Our analysis was based on estimates of the mean trophic level (mTL) of annual landings and the Fishing-in-Balance index (FiB). Food webs of the Channel ecosystem have been altered, as shown by a significant decline in the mTL of fishery landings whilst increases in the FiB index suggest increased fishing effort and fishery expansion. Large, high trophic level species (e.g. spurdog, cod, ling) have been increasingly replaced by smaller, low trophic level fish (e.g. small spotted catsharks) and invertebrates (e.g. scallops, crabs and lobster). Declining trophic levels in fisheries catches have occurred worldwide, with fish catches progressively being replaced by invertebrates. We argue that a network of fisheries closures would help rebalance the trophic status of the Channel and allow regeneration of marine ecosystems.
Molfese, Carlotta; Beare, Doug; Hall-Spencer, Jason M.
A full-cycle laying hen study was conducted to evaluate crab meal (CM) and lobster meal (LM) as feed ingredients for laying hens by assigning four hundred thirty-two 35-wk-old White Leghorns to 1 of 6 diets [control, 2.5% CM, 2.5% LM, 5% CM, 5% LM, and 2.5% CM + 2.5% LM (blend)]. Productive performance and egg parameters were evaluated every 28-d period. Eggs were collected at 67 wk of age from the 5% CM, 5% LM, and blend treatments for analysis of yolk fatty acid composition. At 55 and 67 wk of age, ulnas were collected to determine breaking strength, percent ash, and calcium. Body weights, feed consumption, hen-day production, feed efficiency, and egg quality were not affected (P > 0.05) by treatment. The L* scores of eggs from 5% CM, 5% LM, and blend were lower (P < 0.05) than eggs from the control by the end of the second 28-d period. The a* score of the eggs from 5% CM, 5% LM, and blend was higher (P < 0.05) than the control by the end of the first 28-d period. The content of docosahexaenoic acid (DHA) in egg yolks was different (P < 0.05) among the control (0.17 g/100 g), 5% CM (0.46 g/100 g), 5% LM (0.32 g/100 g), and blend (0.38 g/100 g) treatments. Treatment did not affect (P > 0.05) any of the bone parameters measured at 55 and 67 wk of age. CM and LM supported similar egg production, feed efficiency, egg yolk color, adequate bone strength, and the incorporation of DHA into egg yolks. PMID:22912453
Langille, M A; Anderson, D M; MacIsaac, J L
Ocean surface pH levels are predicted to fall by 0.3-0.4 pH units by the end of the century and are likely to coincide with an increase in sea surface temperature of 2-4 °C. The combined effect of ocean acidification and warming on the functional properties of bivalve shells is largely unknown and of growing concern as the shell provides protection from mechanical and environmental challenges. We examined the effects of near-future pH (ambient pH -0.4 pH units) and warming (ambient temperature +4 °C) on the shells of the commercially important bivalve, Mytilus edulis when fed for a limited period (4-6 h day(-1)). After six months exposure, warming, but not acidification, significantly reduced shell strength determined as reductions in the maximum load endured by the shells. However, acidification resulted in a reduction in shell flex before failure. Reductions in shell strength with warming could not be explained by alterations in morphology, or shell composition but were accompanied by reductions in shell surface area, and by a fall in whole-body condition index. It appears that warming has an indirect effect on shell strength by re-allocating energy from shell formation to support temperature-related increases in maintenance costs, especially as food supply was limited and the mussels were probably relying on internal energy reserves. The maintenance of shell strength despite seawater acidification suggests that biomineralisation processes are unaffected by the associated changes in CaCO3 saturation levels. We conclude that under near-future climate change conditions, ocean warming will pose a greater risk to shell integrity in M. edulis than ocean acidification when food availability is limited. PMID:24489785
Mackenzie, Clara L; Ormondroyd, Graham A; Curling, Simon F; Ball, Richard J; Whiteley, Nia M; Malham, Shelagh K
Persistent bioaccumulative and toxic (PBT) pollutants are chemical contaminants that pose risks to ecosystems and human health. For these reasons, available tissue contaminant data from the US EPA Environmental Monitoring and Assessment Program's National Coastal Assessment were examined to estimate to what areal extent PBTs are found in US estuarine resources. The data document composite, whole-body tissue chemical concentrations for 736 sampling sites across Northeast, Southeast, Gulf of Mexico, and West Coast estuaries. Tissue chemical concentrations were compared to US EPA non-cancer risk guidelines for recreational fishers, because of a lack of ecological guidelines for these chemical concentrations.