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Microalgal blooms: a global issue with negative impact in Chile.

Ecological and health problems posed by microalgal blooms (red tides) occurring in the Southern part of Chile are reviewed. Out of the six human illnesses provoked by microalgal toxins, paralytic shellfish poisoning is the most important, because of its high mortality rate and the high levels of phycotoxins found in contaminated molluscs. Saxitoxin and its analogues bind to a receptor in the voltage-gated sodium channel of neural membranes. The most important toxin-producer microalgae are Alexandrium catenella and Dinophysis acuta. Phycotoxins become concentrated by filter-feeding shellfish, like Mytilus chilensis. Highly sensitive methods available for detection of microalgal toxins are analyzed.

Animals↗

Effects of toxic dinoflagellates and toxin biotransformation in bivalves.

Attempts were made to elucidate the different responses of shellfish to paralytic shellfish poison (PSP) and the PSP donor Alexandrium minutum T1. Five species of edible bivalves (Crassostrea gigas, Meretrix lusoria, Mytilus edulis, Ruditapes philippinarum, and Soletellina diphos) were collected and examined for susceptibility to PSP and PSP donor. It was determined that all five bivalves had low susceptibility to PSP following an intramuscular injection (> 300 MU/20 g). The abnormal effects on bivalves were species-specific and varied with the concentration of A. minutum T1. Judging from the LC50 data (medium lethal concentration), the resistance of bivalves to the toxic dinoflagellate was as follows (least to most resistant): C. gigas < R. philippinarum < M. lusoria < M. edulis, S. diphos. With the exception of S. diphos, the bivalves accumulated very little toxin (< 2 MU/g edible tissue) when they were exposed to 10(7) cells/L of A. minutum for four days. The toxin levels in S. diphos increased with exposure time to the toxic dinoflagellates and accumulated primarily in the digestive gland (88-100%), followed by the gill (0-10%), and other organs (0-8%). Although the concentrations of toxin components in the digestive gland were found to be variable during the exposure period, the toxin profile in the digestive gland of S. diphos during the early exposure period was similar to that of A. minutum. Moreover, toxin components in the gills and in other organs were retained at near constant concentrations during the exposure period.

Animals↗

Creeping eruption caused by a larva of the suborder Spirurina type X.

We report a case of creeping eruption caused by a larva of the suborder Spirurina type X, which developed in a 46-year-old Japanese male. The patient ate small raw squids (Watasenia scintillans) 5 days before the onset of symptoms. On examination, an approximately 25-cm-long serpiginous red track with vesicles was observed from the right to the upper left side of the abdomen of the patient. Histological examination revealed the transverse section of a larval worm in the upper to middle dermis.The patient serum was positive only for the antibody against larvae of the suborder Spirurina type X in ELISA, and negative for all other anti-parasite antibodies. Because a considerable number of people are fond of eating raw or nearly-raw fish and shellfish in Japan, opportunities for developing creeping eruption cause by parasites present in raw fish and shellfish are relatively high.

Animals↗

Human pathogenic protozoa in bivalves collected from local markets in Alexandria.

Bivalves are filter feeders concentrating pathogens from waters and are often eaten raw. Two species, Caelatura Iaronia pruneri and Donax trunculus limiacus commonly known as Gandofli and Om el Kholool respectively were examined. Shellfish were collected from markets in Alexandria to detect any human pathogenic protozoa in them. The infectivity of the detected protozoa was studied by experimental animal inoculation. The results revealed the presence of infectious Cryptosporidia oocysts and Microsporidia spores in both bivalves. Cyclospora sp. was detected only in Gandofli but was non-infectious to mice. There is a potential risk of acquiring protozoa infections if shellfish are eaten raw or insufficiently cooked.

Animals↗

The current position of poikilotherms.

The major research themes in poikilotherm leukaemias, and their progress over the past two years are reviewed. Despite the fact that poikilotherms represent 99% of the animal world including man, background knowledge on most poikilotherm species is sparse. Furthermore, haemic systems in many poikilotherms have functions different to those of homeotherms. Most progress is being made in relation to lethal blood mutant neoplasms in Drosophila, leukaemias of farmed salmonids among the fishes, and among shellfish, the hemic sarcomas of bivalves. The hypothesis that epizootic neoplasia in fish and shellfish populations could be indicative of environmental pollution is being refined in the light of recent studies; environmental xenobiotics are no longer considered to play a primary aetiological role in either lymphomas and leukaemias of fish or haemic sarcomas of bivalves, although xenobiotics may have a primary role in other poikilotherm neoplasms.

Animals↗

[Selected seafoods as the components of a highly nutritional diet].

Shellfish products available on the Polish market were analyzed for the content of protein, fat, carbohydrates, water, ash, energy value and vitamins B. Nutrition value of the analyzed products was determined by analytical methods applying in food chemistry. Concentrations of niacin and vitamin B6 were determined by microbiological methods. The concentration of the selected shellfish in 100 g of the analyzed products and percentage of realization on recommended level of nutritive components by analyzed products for adults were determined.

Animals↗

Marine and freshwater toxins.

In a very busy and exciting year, 2005 included First Action approval of a much needed official method for paralytic shellfish toxins and multiple international toxin symposia highlighted by groundbreaking research. These are the first-year milestones and activities of the Marine and Freshwater Toxins Task Force and Analytical Community. Inaugurated in 2004 and described in detail in last year's General Referee Report (1) this international toxins group has grown to 150 members from many regions and countries. Perhaps most important they are now making important and global contributions to food safety and to providing alternatives to animal-based assays. Official Method 2005.06 was first approved in late 2004 by the Task Force and subsequently Official First Action in 2005 (2) by the Methods Committee on Natural Toxins and Food Allergens and the Official Methods Board. This nonproprietary method (3) is a precolumn oxidation, liquid chromatographic method that makes good use of fluorescence detection to provide high sensitivity detection of the saxitoxins. It has also proven to be rugged enough for regulatory use and the highest level of validation. As pointed out in the report of method principle investigator and Study Director James Lawrence, approval of 2005.06 now provides the first official alternative to the mouse bioassay after many decades of shellfish monitoring. This past year in April 2005 the group also held their first international conference, "Marine and Freshwater Toxins Analysis: Ist Joint Symposium and AOAC Task Force Meeting," in Baiona, Spain. The 4-day conference consisted of research and stakeholder presentations and symposium-integrated subgroup sessions on ciguatoxins, saxitoxin assays and liquid chromatography (LC) methods for saxitoxins and domoic acids, okadaiates and azaspiracids, and yessotoxins. Many of these subgroups were recently formed in 2005 and are working towards their goals of producing officially validated analytical methods. (Abstracts from the Baiona 2005 meeting cited in this report can be found in the online version of the conference abstract book in the Files and Folders section of the Marine and Freshwater Toxins online community at www.aoac.org.) An active topic for discussion in Baiona and subsequent Task Force activities was the expert consultation for Codex which met in Oslo, Norway in 2004 (previously described and cited in last year's GR report, ref 1). The consultation group's executive summary report (http://www.fao.org/es/ESN/food/risk_biotoxin en.stm) describes suggested changes in action levels as well as methods, method validation, and other issues. September 2005 saw the AOAC Task Force efforts further supported by another symposium, "Marine and Freshwater Toxins: Quality Methods for Food Safety and International Trade," at the AOAC INTERNATIONAL Annual Conference in Orlando, Florida. The multidisciplinary talks at this full day symposium ranged from ciguatoxins to cyanobacterial toxins, and spanned toxicology, biochemistry, molecular biology and analytical chemistry. Again, the symposium preceded Task Force meetings. Toxin subgroups, including a new group on cyanobacterial toxins, met for engaging and productive subgroup discussions. All of these activities were preceded by a Wiley Award symposium for Task Force member Mike Quilliam of NRC Canada. These talks, presented at a half-day symposium on the first day of the Annual Meeting, focused on Quilliam's work with LC tandem mass spectrometry (LC/MS/MS) and certified reference standards and materials, and included related presentations by some of his many research collaborators. To maintain flow and continuity between symposia and between Task Force meetings, the group now uses new electronic discussion forums. Individual subgroup areas, under the Marine and Freshwater Toxins Task Force, comprise this online community. First introduced by AOAC INTERNATIONAL in early 2005, these new resources are being used to distribute information and to supplement the in-person subgroup meetings and electronic mail in the group's validation efforts.

Animals↗

Determination of domoic acid in seafoods and in biological tissues and fluids.

Domoic acid is extracted from mussel tissue using the Association of Official Analytical Chemists procedure for paralytic shellfish toxins. This involves boiling the sample for 5 min with O.1N HCl then cooling and centrifuging. An aliquot of the supernatant is diluted 10 to 100 times with water, filtered and analyzed by reversed-phase liquid chromatography with a mobile phase of acetonitrile-water (12:88) at pH 2.5 and absorption detection at 242 nm. The detection limit is about 1 mg/kg domoic acid in seafood samples. The method was successfully used in collaborative studies and a survey of 44 different commercially purchased shellfish products from areas outside of Prince Edward Island showed no domoic acid greater than 1 mg/kg. The same method was applied to urine and feces from monkeys and blood (serum) from humans. The method was unsuccessful for urine and blood which required additional cleanup before analysis. The method worked well for feces at domoic acid levels greater than 1 mg/kg.

Animals↗

Human viruses in sediments, sludges, and soils.

Recent studies have provided a greater understanding of the movement of viruses in the environment by their attachment to solids. These studies have focused on solids-associated viruses present in wastewater discharged into the ocean and on viruses in sludge and wastewater that may be retained in soil following their land disposal. Such ocean or land disposal of wastewater and sludge may result in a discharge of one or more of 120 human enteric virus pathogens including those causing poliomyelitis, viral hepatitis A and acute gastroenteritis.Solids-associated viruses in effluents discharged into coastal waters accumulate in bottom sediments, which may contain 10 to 10 000 more virus per unit volume than the overlying seawater. Solids-associated viruses resuspended by water turbulence may be transported from polluted to distant non-polluted recreational or shellfish-growing water. Transmission of viruses causing hepatitis or gastroenteritis may result from contact by bathers or swimmers with these viruses in recreational waters, or from ingestion of raw or improperly cooked shellfish in which the solids-associated virus had been bioaccumulated.The land disposal of sludge and wastewater has a potential of causing infections in farm workers, contamination of crops, pollution of raw potable water sources or infiltration of ground water. Viruses retained on soils can be released by rain water and may contaminate ground water through lateral and vertical movements.

Adenoviridae↗

Food-borne parasitic infections--old stories and new facts.

A review is presented of food-borne parasitic infections. Parasitic infections with public health hazards both from conventional slaughter animals and from seafood (fish and shellfish) are discussed. The former category includes cysticercosis, echinococcosis, and trichinosis, the latter category covers various trematode, cestode, nematode, and possibly also protozoan infections. Examples of trematode infections are heterophyidiasis, transmitted to man by marine fish, and Paragonimus spp parasites, transmitted by crustaceans. Cestode infections include diphyllobothriasis transmitted by both fresh water fish and fish from brackish waters. Special attention is drawn to the condition known as sparganosis. Of the nematode infections, the eosinophilic granulomatous enteritis due to the genera Anisakis, Phocanema, and Contracaecum, transmitted to man by either marine fish or crustaceans, is mentioned. Two other nematode infections. Angiostrongylus cantonensis and Capillaria philippinensis, can also be transmitted to man by marine fish. Free living amoebae (a.o. Naegleria) may be transmitted to man via shellfish as vehicles. Apart from a possible direct effect of these parasites, chemical alterations in seafood resulting from the presence of parasites may also be deleterious to the consumer. Special attention is drawn to a newly developed serological detection method, the enzyme-linked immunosorbent assay, which makes detection of infection possible not only at the slaughterhouse but also at the farm or in large herds. Strategies to control parasitic infections both in conventional slaughter animals and in seafood are discussed.

Animals↗

Case control study of risk factors for hepatitis A: Naples 1990-1991. Hepatitis Collaborating Group.

An increased incidence of hepatitis A was observed in Naples from 1990-1991. A hospital-based case-control study was carried out to evaluate the relative importance of various risk factors. A hundred and ninety-eight cases and 238 hospital controls were recruited during the study period. The strongest association was contact with a jaundiced person among children. A correlation was also shown for children and adults with raw shellfish consumption and pre-school nursery attendance or presence in the household of children attending pre-school nurseries. History of travel and intravenous drug use were risk factors for adult subjects. Considering the relative importance of the specific risk factors we found that 38% of the acute hepatitis A cases were attributable to contact with a jaundiced person, 15% to presence in the household of children attending pre-school nurseries and 28% to raw shellfish consumption.

Acute Disease↗

[Algal toxins, inhibitors of serine/threonine phosphatases].

Under certain environmental conditions, marine and freshwater phytoplankton may produce phycotoxins inhibitors of serine/thréonine protein phosphatases 1, 2A and 3. In the marine environment, dinoflagellates produce fatty polyethers: okadaic acid and its derivatives, the dinophysistoxins, which accumulate in shellfish and can cause diarrhetic shellfish poisoning (DSP) when ingested. In freshwater, the toxins are microcystins and nodularin, 7 or 5 amino acid cyclic peptides and are hepatotoxic. These toxins have caused massive poisoning of wild animals or domestic livestock and now are a health threat for humans through use of drinking and recreation water. Moreover, all these toxins are potent tumor promoters but belong to a new class, different from the TPA class, because they do not act on Protein Kinase C. Although the mutagenicity Ames test responds negatively, several results show their genotoxic potential, and therefore they are a health hazard through chronic exposition to low doses. Finally, okadaic acid, through its easy penetration in all cellular types can be used as a tool to study mechanisms involved in protein phosphorylation/dephosphorylation processes.

Animals↗

Studies on azaspiracid biotoxins. III. Instrumental validation for rapid quantification of AZA 1 in complex biological matrices.

Azaspiracids are neurotoxins produced by marine algae that have been detected in harvested mussels since 1995. They pose a significant threat to human health through the consumption of contaminated shellfish, and negatively impact the economy of areas where shellfish are harvested and processed. Regulatory agencies are beginning to advocate instrumental assays over traditional mouse bioassay methods. The development and validation of an assay method for AZA 1, the predominant azaspiracid toxin, and the production of a calibration standard and reference material will therefore be vital for quality control in monitoring laboratories worldwide. This report demonstrates a rapid and reproducible liquid chromatography/mass spectrometry (LC/MS) method for separation of all twelve known azaspiracids. Using a triple-quadrupole mass spectrometer, ultra-high sensitivity was obtained at the low-femtogram level injected on-column. At the same time, a linear response of three orders of magnitude was observed. We compared the results with those measured on an ion-trap mass spectrometer. The triple-quadrupole instrument was more sensitive, reliable and reproducible than the ion-trap instrument. The detection limit obtained on the ion-trap mass spectrometer was ten times higher than that obtained on the triple quadrupole. During the study, a new azaspiracid analog (AZA 7c) was discovered.

Animals↗

Classification of Vibrio vulnificus strains by the carbohydrate composition of their capsular polysaccharides.

Pathogenic bacteria are often classified on the basis of the complex polysaccharides found on the surface, usually capsular polysaccharides or lipopolysaccharides. It is common in clinical practice to use reactivity with antisera specific to the various cell surface carbohydrates for this purpose. In this work, we describe a chemotyping method for bacterial capsular polysaccharides which is based on a carbohydrate analysis of an acid hydrolysate of the capsule. High-performance anion-exchange chromatography at high pH (HPAE) with electrochemical detection, which is used for analysis of the hydrolysate, shows preferential sensitivity for sugars. A single acid hydrolysis condition is chosen for screening a large collection of bacterial isolates and a computerized autosampler is used to make possible a large number of rapid analyses. This procedure does not yield a quantitative carbohydrate analysis for the sample but produces a fingerprint which can be used to discriminate among isolates which have different capsular polysaccharide structures. The procedure has been applied to a collection of 120 isolates of Vibrio vulnificus, a water-born species common in shellfish which causes septicemia in immunocompromised individuals, most often from eating of raw oysters. The collection of bacterial isolates includes strains from both clinical cases of septicemia and from such environmental sources such as sea water, sediments, and shellfish. Our results show that a number of unusual sugars including many amino sugars are found in these polysaccharides and that a wide variety of capsular carbotypes in V. vulnificus may be readily distinguished by the HPAE fingerprint.

Anion Exchange Resins↗

Documenting temporal trends of polychlorinated biphenyls in the environment.

In the 15 years since the regulation of polychlorinated biphenyls (PCBs) in the United States, numerous studies have been undertaken to assess PCB levels in the environment. This report reviews the available data from published literature and government agency reports on temporal trends of PCB levels in various environmental compartments. Adequate data were identified to discuss PCB trends in foods for human consumption, human adipose tissue, human blood sera, various fish species, and shellfish. The review found that PCB levels in the human diet currently are less than 1% of the levels detected in the early 1970s as measured by the nationwide Total Diet Study of the FDA. Similarly, EPA data indicate a decrease in the percentage of persons with over 1 ppm of PCBs in adipose tissue from 62% in 1972 to 2% in 1984 when the study was terminated. Studies by the National Oceanic and Atmospheric Administration (NOAA) and other groups have shown a 2- to 10-fold decline in PCB contamination in fish and shellfish from the early 1970s to the late 1980s. The most dramatic declines across all environmental compartments appear in the late 1970s to the mid-1980s which corresponds to the time frame when regulatory controls were imposed by the Toxic Substances Control Act (TSCA). These declines are expected to continue but at slower rates.

Animals↗

Aquatic biotoxins: design and implementation of seafood safety monitoring programs.

Naturally occurring toxicants are usually odorless, tasteless, and generally undetectable by any simple chemical test. Various programs have been established that are effective in reducing risks associated with these toxicants in food. These programs include setting regulatory limits, monitoring susceptible commodities for toxin levels, and using decontamination procedures. Bioassays have been used traditionally to monitor suspect products. All traditional bioassays, however, have one common disadvantage, i.e., the lack of specificity for individual toxins. The lack of available reference standards for specific toxins has also hampered implementation of monitoring programs. Utilizing the knowledge gained with regulatory monitoring and decontamination programs for other toxins, e.g., aflatoxin, similar seafood safety programs can be developed for aquatic biotoxins that will reduce risks and hazards associated with the contaminant to practicable levels and help to preserve an adequate food supply. Research is needed in several areas identified in this article. International cooperation has an important role in achieving these essential elements. Global programs will help in the adequate management of risks associated with aquatic biotoxins. To have an effective monitoring program, it is necessary to define precisely the local needs for information in a short or long time range. It is necessary to have basic knowledge about the biological, chemical, and physical conditions as well as temporal and geographic variations within the region of interest (2). Regardless of the overall success of fish/shellfish toxin monitoring plans, emergencies will occur. Therefore, contingency plans should be developed so there will be no misunderstanding of what actions to take (148). In general, however, the structure of the program must be kept as simple as possible to facilitate fast and uncomplicated flow of information among the various organizations and individuals involved (2). Public health and safety requires the removal of any toxic shellfish from the market, within practicability, and closure of any suspect harvest area. It should be important to remember that economic value of the fish or shellfish resource is always secondary to public health and safety (148).

Environmental Monitoring↗

Viral zoonoses and food of animal origin: caliciviruses and human disease.

Caliciviruses are important veterinary and human pathogens. The viruses gain their name from characteristic cup-shaped structures seen on the virion surface by negative stain electron microscopy. In humans caliciviruses are a major cause of diarrhoeal disease. There are two fundamentally different genome structures amongst human caliciviruses. The Norwalk-like or small round structured viruses (SRSVs) are viruses that have an amorphous structure when viewed by EM, they have a genome composed of 3 major open reading frames (ORFs). These viruses cause epidemic gastroenteritis amongst all age groups. In contrast, the 'classic' human caliciviruses (HuCVs) display the typical calicivirus surface structure and have their capsid ORF fused to and contiguous with the non structural proteins forming one giant polyprotein. HuCVs are predominantly associated with paediatric infections and are only a minor cause of disease in humans. Spread of disease for both SRSVs and HuCVs is usually by faecal oral transmission. SRSVs are a major cause of foodborne gastroenteritis especially linked to the consumption of sewage-contaminated shellfish. However, there is no evidence that these viruses replicate in shellfish or that they originate from an animal source.

Animals↗

[The problem of selective determination of PSP toxins in mussels].

Levels of paralytic shellfish poisoning (PSP) toxins in shellfish are routinely determined by mouse bioassay. In order to improve the qualitative and quantitative determination of PSP toxins, chromatographic techniques with fluorescence detection have been developed. These HPLC methods and the HPLC/MS coupling were used to determine a second PSP toxin which was found, in addition to saxitoxin, in canned Spanish mussels. These canned mussels were rejected in 1986 by the German food control because PSP concentrations were too high. It has been shown that these samples contained mainly dc-saxitoxin.

Animals↗