| Summary: | The presence of <i>Salmonella</i> Enteritidis in poultry houses after cleaning and disinfection can pose a potential risk to public health, as <i>Salmonella</i> remains one of the most important causes of foodborne diseases. This study focused on ten German layer farms (including floor-reared and free-range systems) with a recent history of <i>Salmonella</i> Enteritidis, and samples were collected from July 2018 to March 2021 after the cleaning and disinfection process. A total of 244 swab samples were tested for the presence of <i>Salmonella</i> using real-time PCR, followed by a culture of positive samples. Results revealed that 61 out of the 244 swab samples tested positive for <i>Salmonella</i>, indicating a prevalence of 25% in the samples examined. Among the <i>Salmonella</i>-positive swab samples identified with the PCR assay, 65.6% (40 out of 61) were confirmed by the culture. Of the 40 isolates obtained from the culture, 36 were identified as <i>Salmonella</i> Enteritidis, while 4 were categorized as rough <i>Salmonella</i> strains. This study emphasizes the importance of both the surrounding area of the poultry houses in terms of infection carry-over and the meticulous implementation of cleaning and disinfection procedures to eliminate any remaining infection within the houses. To mitigate the risk of further <i>Salmonella</i> spread on layer farms, additional investigations are recommended to focus on the existing transmission pathways of <i>Salmonella</i> and their genetic diversity.
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