Noroviruses in bivalve shellsh as the acute problem of these days

Uzgoj morskih organizama duž jadranske obale ima dugu tradiciju koja datira još iz 19. stoljeća. Hranidbena vrijednost školjaka se temelji na odnosu bjelančevina i masti, uz velike količine glikogena. Osim velike koristi koju organizmi što žive u vodenoj sredini pružaju čovjeku, oni su i realna opasnost za zdravlje. Školjkaši ltriraju velike količine vode da bi se prehranili, te na takav način kumuliraju različite patogene porijeklom iz ljudskog fecesa. Usvajanjem europske regulative (European regulation 91/492/EC) koja propisuje prihvatljive količine bakterijskih patogena, znatno se smanjio utjecaj bakterija na izbijanje gastroenteritisa, ali ne i bolesti izazvane viralnim uzročnicima. Norovirus (NoV, prije „Norwalk-like virus“) predstavlja najvažnijeg uzročnika nebakterijskog gastro- enteritisa u svijetu. U industrijaliziranom svijetu NoV je moguće odgovoran za preko 80% svih izbijanja gastroenteritisa. NoV spada u porodicu Caliciviridae, a podijeljen je u pet genogrupa (GI-GV), za genogrupe GI, GII i GIV je poznato da izazivaju bolest u ljudi. Nakon infekcije ljudi kontaminiranim školjkašima, epidemija se širi fekalno-oralnim putem, iako do prijenosa bolesti može doći i direktnim kontaktom s osobe na osobu, ili viralnim česticama u aerosolu. Iako je bolest uglavnom blaga i samolimitirajuća, može biti i ozbiljna kod pacijenata koji već imaju nekakve zdravstvene smetnje. Četiri su osnovna problema kod detekcije NoV iz školjaka; niski nivo vi- rusne kontaminacije, velika varijabilnost virusa, prisustvo interferirajućih supstanci koje inhibiraju molekularnu detekciju i genetska varijabilnost NoV. Dva su koraka uključena kod detekcije enterovirusa iz kontaminiranih školjki, a to je izdvajanje i koncentracija vi- rusa iz hepatopankreasa školjkaša i detekcija virusa uz pomoć reverzne transkripcije (RT) PCR metode. Zbog same strategije procjene rizika potrebno je provesti pretraživanje školjkaša na prisustvo NoV. U Evropi (CEN), je osnovao radnu grupu stručnjaka, za razvoj i validaciju referentne horizontalne metode za detekciju NoV u hrani, uključujući školjkaše.Cultivation of marine organisms has a long tradition that dates from the 19th century along the Adriatic coast. Nutritional value of shellfish is based on the proteins and fat, with large amounts of glycogen. Except for the great value of sea organisms, they are also a real danger to human health. Mussels lter large amounts of water to feed, and in that way they bioaccumulate a variety of pathogens, of human faeces origin. By the adoption of the European regulation (European regulation 91/492/EC), which prescribes an acceptable amount of bacterial pathogens, the impact of bacteria on the outbreak of gastroenteritis is significantly reduced, as op- posed to the outbreaks caused by viral pathogens. Norovirus (NoV, “Norwalk-like virus”) is the most important cause of non-bacterial gastroenteritis in the world. In the industrialized world NoV is probably responsible for more than 80% of all gastroenteritis outbreaks. NoV belongs to the family Caliciviridae and is divided into ve genogroups (GI-GV); genogroups GI, GII and GIV are known to cause disease in humans. After infection of people with contaminated shellsh, the epidemic is spreading by fecal-oral route, although the disease can get transmitted by the direct contact from person to person, or by viral particles in aerosol. Gastroenteritis is mostly mild and self-limiting, but it can be serious in patients with some other health problems. There are four basic problems in detection of NoV from shellsh: low levels of viral contamination, high variability of the virus, the presence of substances that inhibit molecular detection and genetic variability of NoV. There are two steps in detection of enteroviruses from contaminated shellfish; the first is extraction and concentration of viruses from the shellfish hepatopancreas and the detection of viruses by reverse transcription (RT) PCR. Due to the risk assessment strategy, analyzing of shellsh for the presence of NoV should be implemented. In Europe (CEN), there was established a working group of experts for the development and validation of reference method for the horizontal detection of NoV in food, including shellsh

Microplastics - a potential risk for seafood safety

Mikroplastika (MP) je prepoznata kao okolišni kontaminant višestrukih opasnosti na temelju fizičkih učinaka zbog veličine čestica, oblika i koncentracije te kemijskih učinaka povezanih s opasnim kemikalijama. Sve je više dokaza o prisutnosti MP u hrani, a posljednjih godina se eksponencijalno povećava i broj publikacija o njezinoj prisutnosti u hrani morskog podrijetla. Međutim, analitičke metode detekcije u hrani morskog podrijetla su unatoč dostupnosti širokog spektra instrumentalnih tehnika za izolaciju i određivanje njezinih fizikalnih svojstava ograničene. Potpuni analitički postupak donosi podatke o kemijskom sastavu i morfološkoj strukturi čestica, ali niti jedna instrumentalna tehnika ne udovoljava svim zahtjevima analize MP. Primjena većeg raspona komplementarnih analitičkih metodologija u pogledu otkrivanja i identifikacije ispunjava zahtjevnost analize ovog okolišnog kontaminanta. Pojava i vrsta unesene MP, kao i njezini zdravstveni učinci koji se trenutno ispituju, uključuju biološke, kemijske i fizičke vrste utjecaja. U svrhu procjene utjecaja MP iz hrane morskog podrijetla na ljudsko zdravlje prvo se mora kvantificirati izloženost, a zatim utvrditi je li ta izloženost dovoljno velika da bi izazvala posljedice sa štetnim učincima. Izloženost ljudi hrani morskog podrijetla izaziva zabrinutosti zbog njezine kontinuirane akumulacije u okolišu. Cilj ovog rada je prikazati dosadašnje spoznaje o prisutnosti MP u hrani morskog podrijetla: školjkašima, ribi i rakovima kao glavnim izvorima. Posebno se analiziraju postojeći analitički pristupi ekstrakcije i instrumentalnog određivanja koncentracije, oblika i vrste materijala MP u zahtjevnim biološkim uzorcima. Istaknuta je potreba daljnje istraživačke aktivnosti o ovom kontaminantu, kao i potreba za stvaranje baze podataka koja je neophodna za uspostavu zakonodavstva u području hrane.Microplastics (MPs) are recognized as a multifaceted stressor based on their physical effects due to particle size, shape, and concentration, and chemical effects ensuing from the use of hazardous chemicals. Evidence of the presence of MPs in food are increasing, and the number of publications on MPs in seafood has increased at an exponential rate in recent years. However analytical detection methods of MPs in seafood are limited even despite the available techniques that enable MP separation and size determination. The analytical detection of MPs requires obtaining information on both the chemical composition and the morphological structure of particles. No single technique is able to chemically and morphologically identify MPs, though a broad range of complementary analytical methodologies have been applied in the detection and identification of MPs. MPs can cause biological, chemical and physical health effects. In order to assess whether the uptake of MPs via seafood can pose a risk to human health, exposure must first be quantified and then it can be determined whether this exposure is high enough to have a detrimental effect. Human exposure to MP contained in food has become a significant concern owing to the increasing accumulation of microplastics in the environment. This paper provides an overview of the existing research on MP presence in seafood, including analytical methods for microplastic separation, and instrumental determination of concentration, shapes, and material types in complex biological material. Regulatory requirements and challenges to support fit-for-purpose research activities on this contaminant are summarized, and information provided for possible regulation on MPs in food

Los hidrocarburos aromáticos policíclicos en cuatro tipos de los jamones croatas

Polycyclic aromatic hydrocarbons (PAHs) represent an important contaminant group in a variety of food products. Since PAHs are commonly present in meat products, this study aimed to determine PAH content in four types of Croatian dry-cured hams (Dalmatian, Drniš, Krk and Istrian) produced by four different processing methods. Determination and quantification of PAHs were performed by High performance liquid chromatography with fluorescence detection (HPLC-FLD). Out of 15 investigated PAHs, 13 compounds were detected. The total average benzo(a)pyrene (BaP) and PAH4 levels obtained from dry-cured hams ranged from 0.05-0.10 µg/kg and 0.41-0.67 µg/kg, respectively. Even though Krk and Istrian dry-cured ham manufacturing processes do not include the smoking phase, no significant differences were found between investigated dry-cured hams in terms of BaP, PAH4, PAH8 and PAH 15 contents. The presence of detected PAHs in non-smoked dry-cured hams could be a result of the addition of spices in the salting phase. BaP and PAH4 contents found in dry-cured ham samples did not exceed the currently legal levels according to the European legislation.Policiklički aromatski ugljikovodici (PAH-ovi) česti su kontaminanti u prehrambenim proizvodima. S obzirom da mesni proizvodi u značajnoj mjeri mogu biti kontaminirani PAH-ovima, cilj ovog rada bio je odrediti njihov udio u četiri vrste hrvatskih pršuta. Istraživanje je provedeno na Dalmatinskom, Drniškom, Krčkom i Istarskom pršutu, čiji se tehnološki procesi proizvodnje razlikuju. Analiza PAH spojeva provedena je primjenom visokoučinkovite tekućinske kromatografije sa flourescentnom detekcijom (HPLC-FLD). Od 15 istraživanih PAH spojeva, identificirano je i kvantificirano 13 spojeva. Prosječna koncentracija benz(a)pirena (BaP) iznosila je 0,05-0,10 µg/kg, dok su prosječne količine PAH4 spojeva iznosile 0,41-0,67 µg/kg. Unatoč tome što proces proizvodnje Krčkog i Istarskog pršuta ne uključuje fazu dimljenja, nisu određene statistički značajne razlike u udjelima BaP, PAH4, PAH8 i PAH15 spojeva između svih istraživanih pršuta. Kontaminacija PAH-ovima kod nedimljenih pršuta je mogla bi biti posljedica primjene začina u fazi soljenja. Koncentracije PAH spojeva određene u uzrocima hrvatskih vrsta pršuta nisu prekoračile dopuštene razine određene trenutno važećim zakonodavstvom Europske Unije.Polyzyklische aromatische Kohlenwasserstoffe (PAH) sind Kontaminate, die in Lebensmitteln häufig erscheinen. Da Fleischprodukte häufig mit polyzyklischen aromatischen Kohlenwasserstoffen kontaminiert sein können, ist das Ziel dieser Arbeit, ihren Anteil in vier Sorten des kroatischen Pršut zu untersuchen. Die Untersuchung wurde am Dalmatiniski, Drniški, Krčki und Istarski Pršut durchgeführt, deren Technologieverfahren sich unterscheiden. Die Analyse der polyzyklischen aromatischen Kohlenwasserstoffe wurde anhand der Hochleistungsflüssigkeitschromatograhpie mit Fluoreszentdetektor (HPLC-FLD) vorgenommen. Von den 15 untersuchten PAH-Verbindungen wurden 13 Verbindungen identifiziert und quantifiziert. Die durchschnittliche Konzentration von Benz(a)pyren betrug 0,05-0,10 µg/kg, während eine durchschnittliche Menge von PAH4 –Verbindungen 0,41-0,67 µg/kg ermittelt wurde. Obwohl das Herstellungsverfahren des Krčki und Istarski Pršut keine Räucherungsphase umfasst, wurden keine statistisch relevanten Unterschiede bei den Anteilen der BaP-, PAH4-, PAH- und PAH15-Verbindungen bei den untersuchten Pršut festgestellt. Die Kontaminierung mit polyzyklischen aromatischen Kohlenwasserstoffen bei nicht geräucherten Pršut könnte die Folge der Verwendung von Gewürzen in der Salzungsphase sein. Die Konzentration der PAH-Verbindungen, die bei den Proben der kroatischen Pršut -Sorten ermittelt wurde, überschreitet nicht die zulässigen Grenzwerte, die durch die aktuell gültige Gesetzgebung der Europäischen Union vorgeschrieben sind.Gli idrocarburi policiclici aromatici (PAH) sono frequenti agenti contaminanti nei prodotti alimentari. Poiché i prodotti a base di carne possono essere significativamente contaminati dagli PAH, questo studio aveva come scopo quello di accertare la loro presenza e la loro percentuale in quattro differenti tipi di prosciutto crudo croato. La ricerca è stata condotta su prosciutti crudi croati differenti (dalmata, di Drniš, dell’isola di Veglia e istriano) i cui processi tecnologici di produzione si differenziano l’uno dall’altro. L’analisi tendente ad accertare la presenza dei composti PAH è stata condotta con la tecnica della cromatografia liquida ad alta prestazione con rivelatori a fluorescenza (HPLC-FLD). Dei 15 PAH oggetto della ricerca, ne sono stati identificati e quantificati ben 13. È stata rilevata una concentrazione media di benzo[a]pirene (BaP) pari a 0,05-0,10 µg/kg, contro una concentrazione media di composti PAH4 pari a 0,41-0,67 µg/kg. Nonostante il fatto che il processo di produzione del prosciutto crudo dell’isola di Veglia e del prosciutto crudo istriano non preveda la fase dell’affumicamento, non sono state accertate differenze statisticamente significative nelle percentuali dei composti BaP, PAH4, PAH8 e PAH15 tra tutti i prosciutti esaminati. La presenza contaminante d’idrocarburi policiclici aromatici nei prosciutti crudi non affumicati potrebbe essere dovuta all’aggiunta di spezie nella fase della salatura. Le concentrazioni di PAH accertate nei campioni dei prosciutti crudi croati esaminati non hanno superato i livelli consentiti previsti dalla legislazione attualmente in vigore nell’Unione Europea.Hidrocarburos aromáticos policíclicos (HAP) son contaminantes frecuentes en los productos alimenticios. Tomando en cuenta que los productos cárnicos pueden estar contaminados en gran medida por los HAP, el fin de este trabajo fue determinar su contenido en cuatro tipos de los jamones croatas. La investigación fue hecha en los jamones de Dalmacia, Drniš, Krk e Istria, con los procesos tecnológicos de producción diferentes. El análisis de los compuestos HAP fue hecho por la cromatografía de líquidos de alta resolución con detección por fluorescencia (HPLC-FLD). Entre los 15 compuestos HAP analizados, 13 fueron identificados. La concentración media del benz(a)pireno (BaP) fue 0,05-0,10 µg/kg, mientras la cantidad media de los compuestos PAH4 fue 0,41-0,67 µg/kg. A pesar de que los procesos de producción de los jamones de Krk y de Istria no incluyen la fase del ahumado, no fueron determinadas las diferencias estadísticamente significativas en el contenido de los compuestos BaP, HAP4, HAP8 y HAP15 en todos los jamones analizados. La contaminación por los PAH de los jamones no ahumados pudo ser la consecuencia del uso de las especies en la fase de salazón. Las concentraciones de los compuestos HAP determinadas en las muestras de los jamones de Croacia no excedieron los niveles determinados por la legislación actual de la Unión Europea

Epizootic Giardiasis in Barn-housed Cattle in Dalmatia

U radu je opisana epizootija giardioze kao uzroka pojave proljeva u mlade teladi. Tijekom trajanja bolesti, parazitološkim i bakteriološkim pretragama pretraženi su uzorci izmeta 21 goveda. Uzročnik je dokazan u izmetu klinički bolesnog teleta te u izmetu 4 odrasla klinički zdrava goveda. Brzo ozdravljenje nakon primjene albendazola potvrđuje nametničku osnovu bolesti. Klinički očitovana giardioza u teladi i supklinička invadiranost odraslih goveda očito su prisutne na području Dalmacije, ali zasigurno nisu dovoljno prepoznate. Cilj je ovoga rada skrenuti pozornost veterinara praktičara na ovu parazitozu kako bi pravodobnim prepoznavanjem i adekvatnom terapijom umanjili štete nastale zbog bolesti i uginuća te posredno sprečavali kontaminacije pašnjaka i nastambi cistama uzročnika.The paper describes epizootic giardiosis as the cause of diarrhoea in young calves. During the time of the disease outbreak 21 cattle stool samples were examined, using parasitological and bacteriological tests. The pathogen was detected in the stool samples of a clinically sick calf and in four clinically healthy adult cattle. The parasitic basis of the disease was confirmed by the cessation of clinical signs of disease after the administration of albendazole. Clinically manifested calf giardiasis and subclinical infestation of adult cattle are obviously present in Dalmatia, but certainly not sufficiently recognized. The aim of this paper is to draw the attention of veterinarian practitioners to this parasitism for timely detection and early treatment to reduce the damage caused by disease and death, and indirectly prevent the contamination of pastures and dwellings by pathogens

Investigation of maedi-visna virus in Dalmatia, using the agar-gel immuno-diffusion test

U Dalmaciji je tijekom 2013. godine provedeno pretraživanje uzoraka seruma ovaca metodom geldifuzijskog precipitacijskog testa radi utvrđivanja pojavnosti virusa maedi-visna. Pretraženo je 408 uzoraka krvnih seruma ovaca i ovnova s područja Dubrovačko-neretvanske, Splitsko-dalmatinske, Šibensko-kninske i Zadarske županije. Cilj je ovoga istraživanja bio po prvi put istražiti seroprevalenciju infekcije virusom maedi-visna u ovaca u Dalmaciji. Infekcija virusom maedi-visna serološki je potvrđena u 66 od 408 pretraženih životinja (16,18 %). Broj pozitivnih životinja u Splitsko-dalmatinskoj županiji iznosio je 24 (20,16 %), u Šibensko-kniniskoj županiji 26 (21,5 %), u Zadarskoj županiji 16 (9,6 %). U Dubrovačkoneretvanskoj županiji nisu pronađeni pozitivni uzorci. Rezultati pretraživanja ujedno su prvi objektivan dokaz o prisutnost protutijela za virus maedi-visna u ovaca u Republici Hrvatskoj. S obzirom na gospodarske štete koje maedi-visna nanosi ovčarstvu, rezultati ovih istraživanja upućuju na potrebu primjene djelotvornih, a prije svega upravnoveterinarskih mjera s ciljem sprečavanja i suzbijanja ove infekcije.In this study we investigated the prevalence of the maedi-visna virus for the first time in Croatia. During 2013 a total of 408 randomly selected sheep and rams blood serum samples were collected from flocks located in four counties in the Dalmatia region, on the eastern coast of the Adriatic Sea. All sera samples were tested using the gel diffusion precipitation test. The presence of maedi-visna virus infection was serologically confirmed in 66 out of 408 tested animals (16.18%). The highest seroprevalence was confirmed in central Dalmatian counties - with seroprevalence of 21.5% in Sibenik-Knin and 20.16% in the Split-Dalmatia County. In the Dubrovnik-Neretva County, in the southernmost part of Dalmatia, no positive samples were found. These results represent the first evidence of maedi-visna virus circulation in sheep in the Republic of Croatia. Given the economic losses that maedi-visna inflicts upon sheepherding, the study results indicate a need for immediate implementation of effective administrative veterinary measures, aimed at preventing and controlling this infection in the Adriatic part of Croatia

Potential Impact of Microplastics and Additives on the Health Status of Loggerhead Turtles (Caretta caretta) Stranded Along the Central Adriatic Coast

AbstractLoggerhead sea turtle (C. caretta) is the official European bioindicator of marine litter in the Mediterranean Sea. In 2019, 8 sea turtles, out of 28 specimens loggerhead on the Adriatic coast of Molise, were subjected to necropsy. The intestinal contents were collected and the microplastics until 0.45 μm were extracted. Qualitative and quantitative assessments were performed by stereomicroscope observation and spectroscopic analyses (attenuated total reflection-Fourier transform infrared spectroscopy, ATR-FTIR and Raman microspectroscopy, RMS). Moreover, the analytical quantification of polyethylene terephthalate (PET), polycarbonate (PC), para phthalic acid (PTA) and bisphenol A (BPA) in fat and liver tissues was performed by LC-MS/MS. Microparticles ranging from 0.45 μm to 1 mm were found in all turtles, for a total of 623, while plastic litter greater than 1 mm were found only in 4 specimens (ranging from 0.03 to 0.11 g). Nineteen different polymers and 10 pigments, including polyester (100% of animals), high-density polyethylene (50%) and polypropylene (50%) were identified. BPA, PTA and PET were detected in fat and liver tissues of all animals, while PC was found only in 50%. A major prevalence was registered in the abdominal fat tissue, although only PC compounds were significantly higher in abdominal tissue (p < 0.05), except for free PTA with liver tissue being the most contaminated (p < 0.05). Microplastics and additives surely impact the health status of turtles that showed gastrointestinal impairment and an important level of contamination in tissues. Graphical abstrac

The Influence of Surface Mycobiota on Sensory Properties of &ldquo;Istarski pr&scaron;ut&rdquo; and &ldquo;Dalmatinski pr&scaron;ut&rdquo;

This study aimed to identify surface mould species overgrowing the Croatian protected meat products &ldquo;Istarski pr&scaron;ut&rdquo; and &ldquo;Dalmatinski pr&scaron;ut&rdquo; and their effect on sensory properties. Dry-cured hams were produced in 2018/2019 and obtained from annual fairs. The predominant surface species found on &ldquo;Dalmatinski pr&scaron;ut&rdquo; were Aspergillus chevalieri, Penicillium citrinum and Aspergillus cibarius, whereas those overgrowing &ldquo;Istarski pr&scaron;ut&rdquo; were Aspergillus proliferans, P. citrinum and Penicillium salamii. The results show species diversity, higher presence, and greater variety of Aspergillus species in &ldquo;Dalmatinski pr&scaron;ut&rdquo; in comparison to &ldquo;Istarski pr&scaron;ut&rdquo;, and significant variations in 9 of 20 sensory attributes. Principal component analysis revealed a clear distinction between the two, and a large contribution of P. salamii and Penicillium bialowienzense to one principal component. The texture traits, smoky odour, muscle and subcutaneous fatty tissue colour, and mould species found are valuable for product characterisation. The results also indicate that mould species may be responsible for some sensory traits, such as tenderness, juiciness, and lesser freshness