Characterization of Vibrio Populations from Cultured European Seabass and the Surrounding Marine Environment with Emphasis on V. anguillarum.

Vibrio species are widely distributed and can be potentially pathogenic to aquatic organisms. In this study, we isolated Vibrio spp. from environmental samples (seawater, sediment, and fish swabs) collected over a three-year period from a fish farm in Mali Ston Bay in the Adriatic Sea, Croatia, and assess their distribution. A total of 48 seawater samples and 12 sediment samples, as well as gill and skin swabs from 110 farmed European seabass, were analysed for the presence of Vibrio. Vibrio strains were identified to the species level by MALDI TOF MS. The analysis revealed that V. alginolyticus was the predominant species in European seabass, followed by V. anguillarum. V. alginolyticus was isolated from the sediments, along with V. gigantis and V. pomeroyi, while V. chagasii, V. cyclitrophicus, V. fortis, V. gigantis, V. harveyi, V. pelagius, and V. pomeroyi were isolated from seawater. V. anguillarum was isolated only twice during two different spring seasons, once from a diseased sea bass and the second time from a healthy sea bass. We analysed these two isolates and found that they differ both genetically and in terms of resistance to antibiotics. Our results confirm the seasonality of vibriosis incidence and the presence of the pathogenic V. anguillarum, which increases the risk of vibriosis.

The Occurrence of Five Unregulated Mycotoxins Most Important for Traditional Dry-Cured Meat Products.

This study investigated the occurrence of 5 unregulated mycotoxins in a total of 250 traditional dry-cured meat products sampled in 2020 and 2021 in five Croatian regions (eastern, northern, central, western, and southern). Aflatoxin B1 (AFB1), ochratoxin A (OTA), sterigmatocystin (STC), citrinin (CIT), and cyclopiazonic acid (CPA) concentrations were related to the geographical region of the product's origin and to local weather. The results revealed the contamination of 27% of samples, namely, STC in 4% of samples in concentrations of up to 3.93 µg/kg, OTA in 10% of samples in concentrations of up to 4.81 µg/kg, and CPA in 13% of samples in concentrations of up to 335.5 µg/kg. No AFB1 or CIT contamination was seen. Although no statistically significant differences in concentrations of individual mycotoxins across the production regions were found, differences in mycotoxin occurrence were revealed. The eastern and western regions, with moderate climate, delivered the largest number of contaminated samples, while the southern region, often compared with subtropics, delivered the smallest, so that the determined mycotoxins were probably mainly produced by the Penicillium rather than the Aspergillus species. Due to the interaction of various factors that may affect mycotoxin biosynthesis during production, the detected concentrations cannot be related solely to the weather.

The emergence of porcine epidemic diarrhoea in Croatia: molecular characterization and serology.

BACKGROUND: Porcine epidemic diarrhoea (PED) is an emergent/re-emergent viral pig disease (caused by the virus belonging to the Coronaviridae family, in specific the Alphacoronavirus genus) of global importance. Clinical presentation is characterized with acute diarrhoea, vomiting and dehydration in pigs of all ages, with a possible high mortality in suckling piglets. The disease emerged in the USA in 2013 causing heavy losses, and re-emerged in Europe in 2014, but with milder consequences. RESULTS: In the spring 2016, PED-like symptoms were reported to be seen on an agricultural holding in Eastern Croatia; laboratory workup confirmed the Croatia's first PED outbreak ever. Porcine epidemic diarrhoea virus (PEDV) strain responsible for the outbreak was of the S-INDEL genotype, much the same as other European PEDV strains. In 2017, a post-outbreak serology was carried out in three counties in Eastern Croatia, revealing seropositivity in pigs bred on four large industrial holdings (9.09%). The seroprevalence across PEDV-positive holdings was up to 82.8%. The latter holdings were unanimously managed by an enterprise that had never reported PED before. CONCLUSIONS: PED has emerged in Croatian pig population causing potentially considerable losses. The circulating strain was of the S-INDEL genotype. Serological workup proved PEDV spread to another four agricultural holdings, demonstrating the importance of not only external, but also internal biosecurity measures.

The impact of frozen storage duration on physical, chemical and microbiological properties of pork.

The aim of the present study was to evaluate the effect of frozen storage duration (3, 6, 9, 12, 15, 18 months) on physical, chemical and microbial properties of pork in three cuts (loin, ham, and belly rib). During frozen storage, significant alterations in physical and chemical parameters were observed through the increase of the total exudate, pH and lightness (L*), and a decrease of shear force and yellowness (b*). Water content in ham samples decreased, while protein content increased. The lipid oxidation (TBA) values in loin and ham samples increased for up to fifteen months of frozen storage, after which they decreased. The proportion of saturated fatty acids in frozen samples was significantly higher than in fresh meat. The total amount of Enterobacteriaceae, S. aureus and Pseudomonas spp. decreased during frozen storage, indicating that freezing may reduce the number of bacteria found in meat. Frozen pork microflora did not change after eighteen months of storage.

Assessment of possible human exposure to ochratoxin A in Croatia due to the consumption of dry-cured and fermented meat products.

Ochratoxin A (OTA) is a toxic secondary metabolite produced by the fungi of Aspergillus and Penicillium species. Data indicate a frequent OTA contamination of cereals and cereal products, and consequently also the contamination of meat and meat products. The aim of this study was to determine a possible level of meat product consumers' exposure to OTA through the consumption of dry-cured and fermented meat products available on the Croatian market. Data showed the weekly OTA intake of 90% of male dry-cured ham consumers to be a maximum of 51.9 ng kg(-1) b.w., i.e., far below the tolerable weekly intake (TWI) of 120 ng kg(-1) b.w. weekly set out by the European Food Safety Authority (EFSA). OTA intake coming from the consumption of other meat products under study is lower and ranges from 0.1 to 42.1 ng kg(-1) b.w. weekly, dependent on the study. The study demonstrated that meat products in Croatia do not constitute a notable source of OTA in the human diet, so that the human health risk coming from the consumption of dry-cured and fermented meat products is negligible.