Powdered formula can be contaminated by several mycotoxins. A multi-mycotoxin UHPLC-MS/MS approach was applied to 83 Italian samples. All samples complied with current regulations, but high prevalence of beauvericin (BEA, 78%) and enniatins (ENNS: B1 77%, B 64%, A1 29%, A 15%) was observed. In vitro assays on BV2 microglial cells treated 24 h with these mycotoxins (0.001-10 µM) showed a significant decrease in cell viability at 10 µM, with increased ROS production and TNF-α release at lower concentrations. When mycotoxins were combined, a significant cell viability reduction was observed (1-10 µM), with increased ROS production at lower concentrations, while no TNF-α increase was detected. In a co-culture model, BV2 cells treated with ENNs and BEA (alone and combined) at 0.1 µM reduced SH-SY5Y neuronal cell viability, except for ENN B1. These results suggest that even low levels, especially under co-exposure conditions, may induce neurotoxic effect mediated by microglia.

This study evaluated the diffusion of Listeria monocytogenes (LM) in a beef steak tartare production chain, aiming to (1) evaluate Listeria spp. diffusion in finishing farms supplying beef cattle, (2) evaluate LM prevalence in carcasses, and (3) map LM diffusion in the production plant. A detection rate of 6/76 was observed in the farm, while carcasses after skinning and before refrigeration tested positive in 19/30 and 11/30, respectively. During tartare production, 57/154 meat and 35/191 environmental samples tested positive. A total of 114 LM isolates were characterized via a whole-genome sequencing approach. Five clonal complexes (CCs) and seven sequence types (STs) were identified, with CC9-ST580 being the most prevalent. Four clusters were identified from both the slaughtering and production phases. Genes related to resistance to fosfomycin, quinolones, sulfonamides, lincosamide, and tetracycline were detected. Two hypervirulent strains (CC6-ST6 and CC2-ST145), harboring a full-length inlA, several virulence genes, and stress islands, were detected. Stress Survival Islet 1 was found in almost all the isolates. The wide diffusion of LM in steak tartare requires the management of some critical phases of the production chain (mainly slaughtering); genomic methodologies could be useful in describing the circulation and virulence of LM strains.

In recent years, there have been important regulatory and management changes in livestock farming in Italy and other European countries. The goal of these changes has been to decrease the use of antimicrobials (antimicrobial stewardship). The objective of this observational study was to explore the potential relationship between the decrease in the use of antimicrobials and the rise in the culling of dairy cattle on 210 farms in the Lodi province. Records of antimicrobials used in dairy herds (measured as the daily defined dose animal for Italy—DDDAit) and culling rates were retrieved from the Italian Ministry of Health official databases. These were statistically analyzed to assess the presence of an association between changes in the DDDAit and changes in the culling rates in 2023 and 2024. The results indicated that the DDDAit exhibited minimal fluctuations over the two-year period (3.61% vs. 2.45%). However, there was a notable shift observed in the culling rates, particularly among smaller herds (24.51 vs. 39.27%). The statistical analysis showed a significant correlation between herd size and antimicrobial treatment, with a higher frequency in larger herds. The results of this study indicate that reducing antimicrobial treatments alone may not necessarily result in better animal welfare or herd sustainability, particularly when it is associated with an increase in the culling rate. It also suggests that antimicrobial treatment rates and culling rates derived from a mandatory recording system provide a simple means of verifying the level of herd health management and of assessing the antimicrobial stewardship approach efficacy.

This study aimed to evaluate the microbiological quality and safety of powdered formulae intended for infant consumption on the Italian market. A total of 83 samples, including 23 infant formulae (PIF), 42 follow-on formulae (FOF), and 18 formulae for special medical purposes (SMPs), were taken between 2023 and 2024. Low total viable counts were highlighted with all SMP samples, 87.0% of PIF samples and 97.6% of FOF samples being compliant with the threshold set by the Code of Hygienic Practice for Powdered Formulae for Infants and Young Children (2.70 Log CFU/g). High contamination levels (>4 Log CFU/g) were found exclusively in PIF (8.7%) and FOF samples (2.4%). Considering potential pathogenic bacteria, the presence of Listeria monocytogenes, Yersinia enterocolitica, Salmonella spp., Bacillus cereus, and Cronobacter sakazakii was investigated. Enumeration of Bacillus cereus, Staphylococcus aureus, and Clostridia was also performed. Only presumptive B. cereus was detected (37 samples, and in 3 samples was enumerated with counts equal to 1 Log CFU/g). A total of 42 presumptive B. cereus isolates were tested for the production of hemolysin BL, phosphatidylcholine-specific phospholipase C, proteases, and for the presence of chromosomal toxin-encoding genes, showing a relevant prevalence of virulence factors and highlighting a potential concern for infants. The antimicrobial resistance pattern of the isolates showed high resistance rates to β-lactams and a moderate resistance to erythromycin. A chemical–physical characterization of the formulae was also performed showing high heterogeneity in terms of pH, Aw, and concentration of organic acids. The results obtained provide useful information for monitoring the potential exposition of infants to microbial populations and to evaluate the safety of the products available on the market.

This study investigated the growth of Bacillus cereus and the production of cereulide toxin in powdered infant formula (PIF) reconstituted according to two different protocols. B. cereus NCTC 11143 spores were inoculated in infant formula samples (approximately 1.5-2 log CFU/ml of final product), that were alternatively reconstituted following the WHO (using water pre-heated to 70°C) or the FDA method (using room-temperature water). Following reconstitution, samples were stored under ideal refrigeration (4°C), thermal abuse (10°C) and strong thermal abuse (20°C) until 72 hours to simulate different storage conditions of leftover formula. B. cereus total count, spore count and cereulide quantification were performed at settled times. Results demonstrated that B. cereus did not grow at 4 and 10°C. At 20°C, spores rapidly germinated and B. cereus grew to very high counts (over 8 Log CFU/ml) in both sample series. Formula prepared according to WHO protocol exhibited earlier growth, exceeding 7 Log CFU/g within 24 hours, whereas such level was reached in 36 h with the FDA protocol. In parallel, cereulide toxin was only detectable after 48 hours of storage at 20°C, when bacterial count exceeded 8 log CFU/g, with lower concentration in the FDA series. These findings suggest the particular nature of the risk posed by B. cereus in PIF if compared to other pathogens, and underscore the critical importance of adhering to proper hygiene, immediate feeding practices, and avoiding the storage of reconstituted formula at abuse temperatures to protect vulnerable infant populations from severe foodborne risks.