Listeriosis is a dangerous zoonosis caused by bacteria of the genus Listeria, with Listeria monocytogenes (LM) being the most pathogenic species. Listeria monocytogenes has been detected in various animal species and in humans, and its ability to evolve from an environmental saprophyte to a powerful intracellular pathogen is driven by the invasion mechanisms and virulence factors that enable cell invasion, replication and cell-to-cell spread. Key regulatory systems, including positive regulatory factor A (PrfA) and the stress-responsive sigma factor σ^B, control the expression of virulence genes and facilitate invasion of host cells. Listeriosis poses a significant threat to cattle, sheep and goat herds, leading to abortions, septicemia and meningoencephalitis, and ruminants are important reservoirs for Listeria, facilitating transmission to humans. Other Listeria species such as Listeria ivanovii and Listeria innocua can also cause disease in ruminants. Resilience of LM in food processing environments makes it an important foodborne pathogen that is frequently transmitted through contaminated meat and dairy products, with contamination often occurring along the food production chain. In humans, listeriosis primarily affects immunocompromised individuals, pregnant women and the elderly and leads to severe conditions, such as meningitis, septicemia and spontaneous abortion. Possible treatment requires antibiotics that penetrate the blood–brain barrier. Despite the relatively low antimicrobial resistance, multidrug-resistant LM strains have been detected in animals, food and the environment. Controlling and monitoring the disease at the herd level, along with adopting a One Health approach, are crucial to protect human and animal health and to minimize the potential negative impacts on the environment.

In this study, the frequency of aflatoxin M1 (AFM1) occurrence in raw milk was investigated across different seasons over a three-year period from 2022 to 2024 in Croatia. Risk assessment was conducted using estimated daily intake (EDI), hazard index (HI), and margin of exposure (MOE) for various age groups and both genders. The frequency of AFM1 detection above the maximum level (ML) ranged from 1.60% to 15.1%. The average incidence of AFM1 exceeding the ML was 5.67%, with the highest incidence recorded in autumn 2024. AFM1 levels within the limit of detection (LOD) and ML were found in 13% of the samples. The average mean value of AFM1 over the three-year period was 19.2 ng/kg. The highest mean AFM1 EDI values were determined for toddlers (0.61–0.67 ng/kg bw/day) and children (0.41–0.43 ng/kg bw/day). The lowest EDI values were observed in elderly females and males (0.058–0.074 ng/kg bw/day). The EDI values for females and males were slightly different. The risk assessment, based on the HI and MOE, indicated that toddlers and children are at the highest risk of exposure to AFM1, which raises significant health concerns. Additionally, consumers of large quantities of milk face a high risk of exposure, particularly during the spring and autumn seasons. For adults and the elderly, milk consumption does not pose a serious health risk.

Melamine is a triazine compound containing 66.6% nitrogen. It is utilised in various industrial applications, including the production of plastics, adhesives, and fertilisers. However, melamine can also contaminate food and animal feed, as it is sometimes added to artificially enhance protein content, which can be toxic to both animals and humans. Notable food contamination incidents involving melamine include the 2007 pet food scandal and the 2008 melamine-contaminated milk scandal in China, which resulted in serious kidney damage and fatalities, particularly among children. The primary health risks associated with melamine arise from its ability to form crystals in the kidneys, potentially leading to renal failure. The body rapidly absorbs and excretes melamine, primarily through urine, as metabolism is minimal. Although acute doses are relatively low in toxicity, long-term exposure, especially through contaminated food, can pose significant health threats. In response to the melamine-related health crises in China, the WHO and EFSA established safe intake levels of 0.2 mg/kg/day for melamine and 1.5 mg/kg/day for cyanuric acid. Various countries have set maximum residue limits for melamine in food, with the United States and China permitting 1.0 mg/kg for infant formula and 2.5 mg/kg for other dairy products. In Europe, the limits are stricter, at 1.0 mg/kg for powdered infant formula, 0.1 mg/kg for liquid formula, and 2.5 mg/kg for foods. The determination of melamine and cyanuric acid in food and feed presents significant challenges. Advanced techniques such as high-performance liquid chromatography and tandem mass spectrometry are the most commonly employed methods.

In this study, the concentrations of metals (arsenic (As), cadmium (Cd), copper (Cu), mercury (Hg) and lead (Pb)), and aflatoxin M1 were analysed in mare's milk samples taken from eight mares over a period of six months during lactation. The results showed that the concentrations of metals in milk occurred in the following order: Cu > Pb > Hg > Cd > As. Key findings of the study included the determined content of metals, depending on lactation stages and animal husbandry practices. Copper had the highest concentration among the metals analysed, while Cd and As were not detected. No statistically significant differences were found in the levels of heavy metals or aflatoxin M1 across different lactation stages. However, statistically significant differences in aflatoxin M1 concentrations were observed depending on the type of animal husbandry, illustrating the influence of environmental and husbandry conditions on milk contamination. The study concludes that there is no toxicological risk for consumers of Croatian Cold-Blooded mare's milk, which is an important finding for food safety and public health. The significance of this study lies in the provision of valuable data on the safety of mare’s milk, especially with regard to its possible contamination with toxic heavy metals and aflatoxin M1. The main objective was to determine the presence and concentrations of the listed contaminants, to evaluate the influence of lactation stage and husbandry, and to assess compliance with established safety standards.

Polycyclic aromatic hydrocarbons (PAHs) are organic compounds consisting of carbon and hydrogen atoms, with more than two fused benzene rings. PAHs are associated with risks to human health, particularly carcinogenesis. One exposure route is the ingestion of contaminated food, which can result from high-temperature preparation and processing (such as grilling, smoking, toasting, frying, and deep-frying), as well as from PAHs present in soil, air, and water due to environmental pollution, including incomplete combustion or pyrolysis of organic matter during industrial processing and motor vehicle exhaust. Deposition of PAHs from the air leads to their entry into soil, which is their main route into the environment – sediments and water. Analytical methods for determining PAHs in environmental samples include extraction, purification, and detection techniques. Sample preparation is a crucial step in the analysis of PAHs, especially in complex food matrices that require efficient extraction and clean-up procedures to isolate PAHs from co-extracted matrix components. This review provides an overview of the available literature – from sample preparation procedures, extraction and clean-up methods, to instrumental analytical techniques in various matrices, including soil, water (wastewater, drinking water, surface water), and food of animal origin (egg, milk) – thus offering a comprehensive summary in one place. The paper also reviews the occurrence of PAHs in different types of matrices in the context of legislation.