The National Prevalence Survey (ENP) of healthcare-associated infections (HAIs) and antimicrobial treatments in long-term care facilities for the elderly (Ehpad) is a cross-sectional survey conducted on a specific day in May–June 2024. Its objectives are to 1) produce prevalence indicators for HAIs and antibiotic treatments (ATB) in nursing homes; 2) engage all healthcare professionals in nursing homes and prescribers in a surveillance project; 3) identify priorities for action regarding infection risk prevention and the appropriate use of antibiotics. Data from 1,288 nursing homes, encompassing 102,166 residents, were analyzed. Residents in 2024 were older (66.6% of residents were 85 years or older in 2024 compared to 63.4% in 2016) than during the previous ENP (Prev’Ehpad) conducted in 2016, with a higher average level of dependency. The prevalence of infected residents (PRI) in 2024 is estimated at 2.35% (95% confidence interval [CI] [2.18–2.53]) and is lower than that measured in 2016 (2.93% [2.57–3.29]). This decrease is even more pronounced after excluding HAIs that were not targeted in 2016 by the Prev’Ehpad survey, such as COVID-19, bacteremias, ENT infections, or fungal infections. With 2,652 HAI cases reported in 2024, the prevalence of infections is estimated at 2.41% [2.23–2.59]. The most common infections are respiratory (36.2%), urinary (31.7%), and skin (25.8%) infections. The microorganisms most frequently isolated from urinary tract infections confirmed microbiologically by cytobacteriological urine examination are, as in 2016, Escherichia coli (53.9%), Klebsiella pneumoniae (11.6%), and Proteus mirabilis (7.5%). The prevalence of residents treated with antibiotics (PRT), estimated at 2.87% [2.74–3.01] in 2024, remains generally stable compared to that of 2016 (2.76% [2.46–3.07]). Nursing homes without a procedure for reevaluating antibiotic therapy, without access to an antibiotic therapy consultant, and without access to hygiene expertise have a higher PRT. Prophylactic treatments are common (22.0%) and on the rise compared to 2016 (13.6%). The main drugs prescribed for prophylaxis are fosfomycin (39.2%), cotrimoxazole (14.4%), amoxicillin (9.8%), and azithromycin (9.1%). The planned duration of curative treatments on an intention-to-treat basis exceeds 7 days for 34.0% of prescriptions, and systematic reevaluation of antibiotic therapy within 3 days is performed for 31.8% of prescriptions. The main drugs prescribed for curative treatment are the amoxicillin/clavulanic acid combination (22.7%), amoxicillin (19.0%), ceftriaxone (14.6%), and cefixime (5.6%). The very high participation of nursing homes demonstrates their commitment to preventing infection risk and the appropriate use of antibiotics. The results provide guidance for actions aimed at strengthening certain practices regarding the diagnostic and therapeutic management of infections, such as evaluating the appropriateness of prophylactic treatments and the systematic reassessment of curative treatments within 3 days.