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The increasing consumption of fish has raised public health concerns regarding fish-borne zoonotic helminths (FBZHs), which are recognized as significant food-borne parasites worldwide. In freshwater environments, Clinostomum complanatum, Opisthorchis felineus, Pseudamphistomum truncatum, Dibothriocephalus latus and Eustrongylides excisus are of particular concern in Italy and neighbouring countries. This study aimed to assess the prevalence of these FBZHs in five commercially and ecologically relevant freshwater fish species from Lake Varese, a heavily anthropized and understudied basin in northern Italy. A total of 59 fish were examined via necropsy and stereomicroscopic inspection of skeletal muscles. Only Eustrongylides spp. larvae were detected, with a prevalence of 16.9%. Molecular analysis (ITS region) identified them as E. excisus. This study reports, for the first time in Western Europe, E. excisus in Sander lucioperca (p = 12.5%) and Esox lucius (p = 8.3%). The highest prevalence occurred in Silurus glanis (p = 37.5%), followed by Perca fluviatilis (p = 25.0%), while Tinca tinca showed no infection. These findings confirm that among the FBZHs considered, E. excisus is currently present and expanding both in host range and geography in Italian lakes, underscoring the need for updated epidemiological data to support risk assessment, food safety and zoonotic parasite control in freshwater fisheries.

Hydroxyanthracene derivatives (HADs) are natural compounds that occur in several botanical species belonging to the genera Aloe L., Cassia L., Rheum L., Frangula Mill., and Rhamnus L. While they are widely used for their laxative effects, concerns persist about their potential genotoxicity and carcinogenicity. This study presents the development, validation, and application of a sensitive and rapid LC-MS/MS method to detect HAD (aloins, aloe-emodin, emodin, and danthron) levels in botanical food products and supplements. The method was validated according to criteria established by the European Union Reference Laboratory for Mycotoxins and Plant Toxins, and Regulation (EU) No 2783/2023 and was demonstrated to be fit-for-purpose. It was applied to 43 samples collected from the Italian market, including beverages, liquid and solid food supplements, herbal infusions, and jam based on aloe, senna, cassia, rhubarb, and frangula. The results revealed that 33% of the analyzed samples contained detectable HAD concentrations above the limit of quantification (LOQ = 0.5 mg/kg). The highest concentrations, up to 1352.9 mg/kg for the sum of aloin A and B, were found in solid food supplements derived from senna, cascara, rhubarb, and frangula. Aloe-emodin reached 213.4 mg/kg in a solid food supplement sample containing the same plant species, while the maximum detected concentration of emodin was 259.7 mg/kg in a senna-based supplement. No sample contained danthron. Conversely, in the majority of aloe beverage samples, HAD levels were found to be below the LOQ. In order to ensure the safety of consumers, it is essential that a more rigorous market surveillance of botanical food products is implemented, along with further toxicological studies.

Among biogenic amines, histamine is most frequently involved in foodborne intoxication. To evaluate histamine formation in tuna, several storage conditions were reproduced. An LC-MS/MS method was used for analytical determinations. Fresh tuna samples (not contaminated and grafted with tuna muscle naturally incurred with histamine at 6000 mg/kg) were stored at 4, 12, and 20 °C, and daily samples were collected for 6 days. The development of histamine was observed only in grafted tuna samples. At 4 °C, histamine formation progressed from 12.8 mg/kg (day 1) up to 68.2 mg/kg (day 6). At 12 °C, higher concentrations developed (23.9 mg/kg on day 1 up to 2721.3 mg/kg on day 6) relative to 20 °C (from 12.0 to 1681.0 mg/kg). It was found that at 4 °C, if grafted tuna was submerged in oil, histamine formation progressed more slowly. In a naturally contaminated sample, it was observed that the histamine distribution was uniform, while the normal cooking process did not affect the histamine level. Furthermore, it was found that the use of histamine-contaminated equipment for food handling may result in histamine formation in food. These results confirm the importance of implementing good hygiene practices and respecting the cold chain.

In a balanced diet, regular fish consumption provides positive outcomes for human health. On the other hand, the seafood supply chain faces a significant food safety risk due to the presence of potentially toxic elements (PTEs). In the present study, to assess the risk for Italian consumers, the concentrations of five PTEs, namely lead, chromium, cadmium, mercury, nickel, and aluminum, were determined in the three most consumed preserved fish in Italy: tuna (Thunnus albacares, Katsuwonus pelamis), mackerel (Scomber scombrus) and anchovy (Engraulis encrasicolus). Samples were collected from the national market, and the instrumental analysis was performed by inductively coupled plasma mass spectrometry (ICP-MS). The analyzed PTEs were found in all the species that were investigated. However, after considering the target hazard quotient (THQ) and the hazard index (HI), it was observed that the three fish preserves did not pose any risk of chronic toxicity for the average consumer, even at the highest concentrations detected. However, for significant consumers, mercury detected in tuna samples represented almost 90% of the tolerable weekly intake (TWI) reported by the European Food Safety Authority (EFSA), representing a matter of concern for consumers, particularly regarding developmental neurotoxicity, whose HI exceeded 111%. The acute toxicity of nickel was also considered for significant consumers at the highest concentration detected, and the margin of exposure (MOE) calculated was above 7000, much higher than the value of 30 indicated by EFSA. Due to the lack of data on non-professional carcinogenicity or human intake through foods with low cancer risk, this toxicity was not considered in the analysis of PTEs.

The increasing consumption of fish has raised public health concerns regarding fish-borne zoonotic helminths (FBZHs), which are recognized as significant food-borne parasites worldwide. In freshwater environments, Clinostomum complanatum, Opisthorchis felineus, Pseudamphistomum truncatum, Dibothriocephalus latus and Eustrongylides excisus are of particular concern in Italy and neighbouring countries. This study aimed to assess the prevalence of these FBZHs in five commercially and ecologically relevant freshwater fish species from Lake Varese, a heavily anthropized and understudied basin in northern Italy. A total of 59 fish were examined via necropsy and stereomicroscopic inspection of skeletal muscles. Only Eustrongylides spp. larvae were detected, with a prevalence of 16.9%. Molecular analysis (ITS region) identified them as E. excisus. This study reports, for the first time in Western Europe, E. excisus in Sander lucioperca (p = 12.5%) and Esox lucius (p = 8.3%). The highest prevalence occurred in Silurus glanis (p = 37.5%), followed by Perca fluviatilis (p = 25.0%), while Tinca tinca showed no infection. These findings confirm that among the FBZHs considered, E. excisus is currently present and expanding both in host range and geography in Italian lakes, underscoring the need for updated epidemiological data to support risk assessment, food safety and zoonotic parasite control in freshwater fisheries.