# Characterization of Infant Formulae Marketed in Italy and Virulence Potential of Bacillus cereus Isolates

## TL;DR

This study analyzed infant formula in Italy, finding some samples had high contamination and Bacillus cereus with potential virulence.

## Contribution

The study provides new insights into the microbial safety and virulence potential of Bacillus cereus in Italian infant formula products.

## Key findings

- Most infant formula samples met microbiological safety thresholds, but some had high contamination levels.
- Bacillus cereus isolates showed significant virulence factors and resistance to β-lactam antibiotics.
- Formulae exhibited high chemical-physical variability in pH, water activity, and organic acid content.

## Abstract

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.