Monitoring Hygiene Protocols and Exploring Alternatives to Counteract Resistant Pathogens: A Case Study from Southern Italy on Healthcare-Associated Infection Control

Healthcare-associated infections (HAIs) remain a major public health concern, contributing to increased morbidity, mortality, and antimicrobial resistance. Healthcare workers (HCWs) are recognized as key vehicles in the transmission of nosocomial pathogens, primarily via contaminated hands and medical devices. This study assessed the effectiveness of hand hygiene protocols among HCWs, their correlation with bloodstream infections, and the potential of natural antimicrobial agents as complementary preventive measures. Between January and June 2025, 128 hand samples were collected from HCWs in surgical, intensive care, and internal medicine units of hospitals managed by ASL Caserta (Marcianise, n = 65; Piedimonte Matese, n = 30; Sessa Aurunca, n = 18; Maddaloni, n = 15). Sampling was performed upon entry to clinical areas and after antiseptic handwashing, using Rodac TSA plates to quantify microbial load (CFU/cm2). Isolates were identified via MALDI-TOF, and multidrug resistance was confirmed using the Phoenix BD system. Microbial growth was detected in 54.7% of samples. Coagulase-negative staphylococci, mainly Staphylococcus epidermidis and S. hominis, accounted for 67.1% of positive cultures, followed by Enterobacteriaceae (28.6%). Comparison with concurrently collected blood cultures revealed potential overlapping pathogens, with Staphylococcus spp. prevalence ranging from 35 to 56% and Gram-negatives from 18 to 39. Selected isolates were further tested for susceptibility to natural antimicrobial agents, derived from hop, red vine leaf, green tea, and pomegranate fruit, as well as thyme essential oil. Thyme essential oil (Thymus vulgaris) demonstrated notable antimicrobial activity, in several cases surpassing that of standard hygiene agents. These findings highlight not only that maintaining high standards of hand hygiene, proper care of invasive devices, and continuous microbiological surveillance is critical for preventing HAIs, but also that incorporating natural antimicrobial compounds into hygiene protocols may provide an effective and sustainable adjunct to reduce microbial contamination and combat infections caused by multidrug-resistant organisms.