Phycotoxins in marine organisms - a potential threat to consumer safety

Fikotoksini predstavljaju skupinu prirodnih kemijskih kontaminanata u koje spadaju azaspiracidi (AZA), okadaična kiselina (OA) i njeni derivati, pektenotoksini (PTX), jesotoksini (YTX), ciklički imini (CI), domoična kiselina (DA), saksitoksini (STX), brevetoxini (BTX), ciguatoksini (CTX) i palitoksini (PlTX). Rezultati dosadašnjih istraživanja ukazuju na učestalu pojavnost fikotoksina u morskim organizmima, a poznato je da ove tvari mogu imati štetan učinak na okoliš i zdravlje ljudi, budući da su pored akutne toksičnosti pojedini ujedno i tumor promotori, odnosno da već i u malim koncentracijama pokazuju mutageno i toksično djelovanje u organizmu. Stoga su s ciljem zaštite zdravlja potrošača zakonodavstvom određene najveće dopuštene količine (NDK) fikotoksina u školjkaša, bodljikaša, plaštenjaka i morskih puževa (EZ 853/2004; EU 786/2013), iako neki od njih, poput cikličkih imina, još nisu uključeni u programe monitoringa, odnosno za njih legislativom nije određena NDK. U daljnjim istraživanjima, uz primjenu potvrdnih visokospecifičnih analitičkih metoda u ispitivanjima pojavnosti fikotoksina, potrebno je ispitati njihove učinke na osjetljiviji dio populacije te zbog mogućeg sinergičkog učinka u organizmu ustvrditi i supojavnost, kao i provesti procjenu izloženosti potrošača zbog prisustnosti ovih toksičnih tvari u različitim morskim organizmima namijenjenih konzumaciji. U ovom radu dan je pregled literaturnih spoznaja o svojstvima najznačajnijih predstavnika fikotoksina, njihovim toksičnim učincima u organizmu i pojavnosti u različitim vrstama morskih organizama.Phycotoxins are natural chemical con- taminants that include azaspiracids (AZAs), okadaic acid (OAs), pectenotoxins (PTXs), yessotoxins (YTXs), cyclic imines (CIs), domoic acid (DA), saxitoxins (STXs) breve- toxins (BTXs), ciguatoxins (CTXs) and palyt- oxins (PlTX). The results of previous research have indicated the frequent occurrence of phycotoxins in marine organisms, and it is known that these substances can have a det- rimental effect on both the environment and human health, since in addition to acute tox- icity, some are tumour promoters, i.e., they show mutagenic and toxic effects in the body even when present in low concentrations. Therefore, in order to protect consumers, the legislation has set maximum levels (MLs) for phycotoxins in bivalve molluscs, echino- derms, tunicates and marine gastropods (EC 853/2004; EU 786/2013), although some, such as cyclic imines, have not yet been included in monitoring programmes and MLs have not yet been determined by the legislation. In further research, using confirmatory highly specific analytical methods to test for the oc- currence of these substances, it is necessary to examine their effects on a more sensitive part of the population due to possible syn- ergistic effects in the body to determine co-occurrence, and to assess consumer exposure due to the presence of these toxic substances intended for consumption. This paper pro- vides an overview of the literature on the properties of the most important representa- tives of phycotoxins, their toxic effects in the body and their occurrence in different types of marine organisms

Halophilic vibrios in shellfish as potentially pathogenic causative agents of zoonoses in humans

Uzgoj, izlov i promet školjkašima reguliran je brojnim zakonima kojima je cilj stavljanje na tržište zdrave hrane. Zakonodavstvo ne predviđa kontrolu Vibrio spp., što može rezultirati stavljanjem na tržište i konzumacijom školjkaša kontaminiranih bakterijama ovog roda te posljedično prouzročiti pojavu bolesti u ljudi. U radu je prikazano značenje najvažnijih potencijelno patogenih halofilnih vibrija poput V. cholerae, V. parahaemolyticus i V. vulnificus u infekciji ljudi. Tijekom zadnjih dvadesetak godina u zemljama mediteranske regije provedena su istraživanja, koja su redom potvrdila prisutnost vibrija u morskoj vodi, školjkašima, sedimentu i u otpadnim vodama. U Hrvatskoj je u rijetkim istraživanjima školjkaša dokazana duž Jadranske obale prisutnost Vibrio spp., uz dominaciju vrste V. parahaemolyticus. Sve izraženija pojava globalnog zatopljenja podudara se i s neočekivanim pojavama infekcija s Vibrio spp. na sjeveru Europe, ponajprije na području Baltičkog mora. Četiri su važne značajke pripadnika roda Vibrio koje ih čine pogodnima u procjeni klimatskih promjena. To su: osjetljivost na temperaturu (jako dobro rastu pri temperaturama iznad 15 °C), brzo umnažanje, pojava u područjima gdje se nikada nisu javljali (kao što su sjeverna Europa, Aljaska, Čile, sjevero - istočno područje SAD) i mogućnost ulaska u organizam putem hrane i izlaganjem kontaminiranoj vodi. S gledišta sigurnosti hrane i školjkaša kao hrane, globalno zatopljenje i klimatske promjene imaju značajan utjecaj na dinamiku i rasprostranjenost potencijalno patogenih halofilnih vibrija na područja u kojima nisu povijesno dokazani kao uzročnici oboljenja ljudi. Posljednjih su desetljeća razvijane različite metode s ciljem pouzdanije identifikacije Vibrio vrsta. Radilo se na poboljšanju klasičnih mikrobioloških metoda, a većina je istraživanja usmjerena prema razvoju i primjeni molekularnih tehnika. Istraživanjima kojima su uspoređivani rezultati kulturelnih mikrobioloških metoda i lančane reakcije polimerazom (PCR), dokazana je značajno veća osjetljivost i pouzdanost molekularnih metoda u dokazu uzročnika.Breeding, collection and trade of bivalve molluscs is regulated by a number of laws aimed at placing healthy food on the market. The legislation does not provide for the control of Vibrio spp., which may result the marketing and consumption of bivalve molluscs contaminated with Vibrio spp. bacteria, ultimately causing human disease. The significance of the most important potentially pathogenic halophilic vibrios, such as V. cholerae, V. parahaemolyticus and V. vulnificus, in human infection is presented in this paper. Over the last twenty years, research has been conducted in the countries of the Mediterranean region, confirming the presence of vibrios in seawater, shellfish, sediment and wastewater. In Croatia, the presence of Vibrio spp. has been proven in rare studies along the Adriatic coast, with the dominance of V. parahaemolitycus in shellfish. The increasingly pronounced phenomenon of global warming coincides with the unexpected occurrence of Vibrio spp. infections in northern Europe, primarily in the Baltic Sea. Four important features of the members of the genus Vibrio make them suitable for assessing climate change. These are temperature sensitivity (very good growth at temperatures above 15°C); rapid replication; occurrence in areas where they have never occurred (such as northern Europe, Alaska, Chile, northeast USA), and the possibility of entering the body through food consumption and exposure to contaminated water. From the point of view of food and shellfish safety as food, global warming and climate change have a significant impact on the dynamics and distribution of the potentially pathogenic halophile vibrios in areas where they have not historically been proven to cause human diseases. In recent decades, various methods have been developed with the aim of more reliable identification of Vibrio species. The aim was to improve classical microbiological methods, while numerous studies were based on the development and application of molecular techniques. Studies comparing the results of culture microbiological methods and polymerase chain reaction (PCR) have proven to have significantly higher sensitivity and reliability in the detection of pathogens

With food to health : proceedings of 11th International symposium

Proceedings contains 13 original scientific papers, 10 professional papers and 2 review papers which were presented at "10th International Scientific and Professional Conference WITH FOOD TO HEALTH", organised in following sections: Nutrition, Dietetics and diet therapy, Functional food and food supplemnents, Food safety, Food analysis, Production of safe food and food with added nutritional value

With food to health : proceedings of the 10th International scientific and professional conference

Proceedings contains 13 original scientific papers, 10 professional papers and 2 review papers which were presented at "10th International Scientific and Professional Conference WITH FOOD TO HEALTH", organised in following sections: Nutrition, Dietetics and diet therapy, Functional food and food supplemnents, Food safety, Food analysis, Production of safe food and food with added nutritional value

Occurrence and Seasonal Monitoring of Domoic Acid in Three Shellfish Species from the Northern Adriatic Sea

As filter feeders, bivalves and ascidians can accumulate contaminants present in the environment and pass them on to higher food chain levels as vectors. The consumption of bivalves contaminated with the potent neurotoxin domoic acid (DA) can cause amnesic shellfish poisoning in humans. The aim of this study was to determine seasonal differences in occurrence and accumulation of this phycotoxin in European oysters (Ostrea edulis Linnaeus, 1758) (n = 46), Queen scallops (Aequipecten opercularis Linnaeus, 1758) (n = 53), and edible ascidians of the Microcosmus spp. (n = 107), originating from the same harvesting area in the Northern Adriatic Sea. The quantification was performed using ultra-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) preceded by derivatization with dansyl chloride. DA was found in very low concentrations throughout the year, with a maximum value of 810 μg/kg in Queen scallops. This study reveals differences in the occurrence and accumulation of DA between Queen scallops and the other two investigated species (oysters and ascidians) and the highest concentrations during the colder part of the year. Even though DA was detected in all of them, Queen scallops showed higher DA accumulation compared to the other two (p < 0.001), hence representing a sentinel species suitable for the monitoring of DA level in seafood

Profile of phycotoxins in seafood from the Istrian peninsula waters and exposure assessment for Croatian consumers

Plodovi mora važan su dio uravnotežene prehrane zbog velike nutritivne vrijednosti i dobrobiti za zdravlje. Neki od njih, poput školjkaša i mješčićnica, zbog specifičnog načina hranidbe i filtracije velikih količina vode u svom tkivu mogu nakupljati različite kontaminante, poput fikotoksina, te na taj način postaju njihovi vektori u prehrambenom lancu. U ovom istraživanju validirane su LC-MS/MS metode za određivanje domoične kiseline u tragovima i lipofilnih toksina, uključujući slabo istražene cikličke imine i LC-FLD metoda za određivanje paralitičkih toksina, te su primijenjene za ispitivanje profila ovih skupina fikotoksina u uzorcima dagnji (n = 416), kamenica (n = 104), kapica (n = 52) i mješčićnica (n = 104). Od ukupnog broja uzoraka, u njih šest kvantificirani su fikotoksini u koncentraciji većoj od NDK. U najvećem broju uzoraka kvantificirani su SPX1 i GYM, zatim YTX, DA, OA, PnTX-G, AZA2, GTX2,3, GTX1,4, AZA1 i PTX2, uglavnom u niskim koncentracijama. Temeljem utvrđenih koncentracija i prehrambenih navika utvrđeno je da potrošači nisu izloženi akutnom toksičnom učinku većine fikotoksina obuhvaćenih istraživanjem, no utvrđena je izloženost akutnom štetnom djelovanju okadaične kiseline i paralitičkim toksinima. Utvrđena je statistički značajna (p < 0,05) sezonska pojavnost fikotoksina, utjecaj lokaliteta, kao i razlika u pojavnosti među vrstama izloženim jednakim okolišnim utjecajima.Due to its high nutritional value and health benefits, seafood is an important part of a healthy diet. Due to their specific way of feeding, bivalve mollusks and ascidians filter large amounts of water and may accumulate many contaminants in their tissues, such as phycotoxins, thus becoming their vectors in the food chain. In the framework of this dissertation, LC-MS/MS methods for the determination of domoic acid traces, lipophilic toxins, including insufficiently studied cyclic imines, and LC-FLD method for the determination of paralytic shellfish toxins were validated and applied to determine the profile of these phycotoxins in mussels (n = 416), oysters (n = 104), scallops (n = 52) and ascidians (n = 104). Phycotoxins were quantified in 6 samples in concentrations higher than the MPL. SPX1 and GYM were the most frequent, followed by YTX, DA, OA, PnTX-G, AZA2, GTX2,3, GTX1,4, AZA1, and PTX2, present mostly in low concentrations. Based on the established concentrations and dietary habits of consumers in Croatia, an exposure assessment of consumers to phycotoxins was done. Consumers were not exposed to the acute toxic effect of most of the phycotoxins included in this research, but they were exposed to the acute harmful effect of okadaic acid and paralytic shellfish toxins. Statisticaly significant seasonality (p < 0,05) and influence of geographical region on the occurrence of phycotoxins were observed, as well as the interspecies differences in their accumulation in samples collected from the same region, exposed to the same environmental influence

Profile of phycotoxins in seafood from the Istrian peninsula waters and exposure assessment for Croatian consumers

Plodovi mora važan su dio uravnotežene prehrane zbog velike nutritivne vrijednosti i dobrobiti za zdravlje. Neki od njih, poput školjkaša i mješčićnica, zbog specifičnog načina hranidbe i filtracije velikih količina vode u svom tkivu mogu nakupljati različite kontaminante, poput fikotoksina, te na taj način postaju njihovi vektori u prehrambenom lancu. U ovom istraživanju validirane su LC-MS/MS metode za određivanje domoične kiseline u tragovima i lipofilnih toksina, uključujući slabo istražene cikličke imine i LC-FLD metoda za određivanje paralitičkih toksina, te su primijenjene za ispitivanje profila ovih skupina fikotoksina u uzorcima dagnji (n = 416), kamenica (n = 104), kapica (n = 52) i mješčićnica (n = 104). Od ukupnog broja uzoraka, u njih šest kvantificirani su fikotoksini u koncentraciji većoj od NDK. U najvećem broju uzoraka kvantificirani su SPX1 i GYM, zatim YTX, DA, OA, PnTX-G, AZA2, GTX2,3, GTX1,4, AZA1 i PTX2, uglavnom u niskim koncentracijama. Temeljem utvrđenih koncentracija i prehrambenih navika utvrđeno je da potrošači nisu izloženi akutnom toksičnom učinku većine fikotoksina obuhvaćenih istraživanjem, no utvrđena je izloženost akutnom štetnom djelovanju okadaične kiseline i paralitičkim toksinima. Utvrđena je statistički značajna (p < 0,05) sezonska pojavnost fikotoksina, utjecaj lokaliteta, kao i razlika u pojavnosti među vrstama izloženim jednakim okolišnim utjecajima.Due to its high nutritional value and health benefits, seafood is an important part of a healthy diet. Due to their specific way of feeding, bivalve mollusks and ascidians filter large amounts of water and may accumulate many contaminants in their tissues, such as phycotoxins, thus becoming their vectors in the food chain. In the framework of this dissertation, LC-MS/MS methods for the determination of domoic acid traces, lipophilic toxins, including insufficiently studied cyclic imines, and LC-FLD method for the determination of paralytic shellfish toxins were validated and applied to determine the profile of these phycotoxins in mussels (n = 416), oysters (n = 104), scallops (n = 52) and ascidians (n = 104). Phycotoxins were quantified in 6 samples in concentrations higher than the MPL. SPX1 and GYM were the most frequent, followed by YTX, DA, OA, PnTX-G, AZA2, GTX2,3, GTX1,4, AZA1, and PTX2, present mostly in low concentrations. Based on the established concentrations and dietary habits of consumers in Croatia, an exposure assessment of consumers to phycotoxins was done. Consumers were not exposed to the acute toxic effect of most of the phycotoxins included in this research, but they were exposed to the acute harmful effect of okadaic acid and paralytic shellfish toxins. Statisticaly significant seasonality (p < 0,05) and influence of geographical region on the occurrence of phycotoxins were observed, as well as the interspecies differences in their accumulation in samples collected from the same region, exposed to the same environmental influence

Occurrence and Seasonal Monitoring of Domoic Acid in Three Shellfish Species from the Northern Adriatic Sea

As filter feeders, bivalves and ascidians can accumulate contaminants present in the environment and pass them on to higher food chain levels as vectors. The consumption of bivalves contaminated with the potent neurotoxin domoic acid (DA) can cause amnesic shellfish poisoning in humans. The aim of this study was to determine seasonal differences in occurrence and accumulation of this phycotoxin in European oysters (Ostrea edulis Linnaeus, 1758) (n = 46), Queen scallops (Aequipecten opercularis Linnaeus, 1758) (n = 53), and edible ascidians of the Microcosmus spp. (n = 107), originating from the same harvesting area in the Northern Adriatic Sea. The quantification was performed using ultra-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) preceded by derivatization with dansyl chloride. DA was found in very low concentrations throughout the year, with a maximum value of 810 μg/kg in Queen scallops. This study reveals differences in the occurrence and accumulation of DA between Queen scallops and the other two investigated species (oysters and ascidians) and the highest concentrations during the colder part of the year. Even though DA was detected in all of them, Queen scallops showed higher DA accumulation compared to the other two (p < 0.001), hence representing a sentinel species suitable for the monitoring of DA level in seafood

Variations in nutritive composition of three shellfish species

Nutritive composition, fatty acid profile and health- related lipid indices of natural-born European flat oyster (Ostrea edulis), variegated scallop (Chlamys varia) and smooth scallop (Flexopecten glaber) in 108 samples originating from the Adriatic sea, recovered on a monthly basis were investigated. Out of three shellfish species, the lowest share of saturated fatty acids, the most favourable ratio of polyunsaturated over saturated fatty acids, the most favourable atherogenic and thrombogenic index, and the most favourable ratio of hypocholesterolaemic over hypercholesterolaemic fatty acids were seen in oysters, sampled during springtime. No statistically significant inter- seasonal differences between basic chemical parameters and fat quality indices were established

Variations in nutritive composition of three shellfish species

Nutritive composition, fatty acid profile and health- related lipid indices of natural-born European flat oyster (Ostrea edulis), variegated scallop (Chlamys varia) and smooth scallop (Flexopecten glaber) in 108 samples originating from the Adriatic sea, recovered on a monthly basis were investigated. Out of three shellfish species, the lowest share of saturated fatty acids, the most favourable ratio of polyunsaturated over saturated fatty acids, the most favourable atherogenic and thrombogenic index, and the most favourable ratio of hypocholesterolaemic over hypercholesterolaemic fatty acids were seen in oysters, sampled during springtime. No statistically significant inter- seasonal differences between basic chemical parameters and fat quality indices were established