Background and Objectives: Despite extensive research on the sources and transmission pathways of Hospital-Acquired Infections (HAIs), the role of cleaning staff as potential vectors has been under-explored. This study addresses the gap by examining the cleaning staff’s role in nosocomial infection transmission, focusing on pathogenic bacteria and fungi colonization. Identifying potential pathogens harbored by cleaning staff that carry the risk of causing HAIs, especially methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae. Materials and Methods: This current cross-sectional study was conducted on 96 cleaning staff at King Fahad Hospital of the University and Family and Community Medicine-Centre, Eastern Province, Saudi Arabia. Sample collection targeted hands and nostrils using cotton swabs, followed by laboratory testing, including MALDI-TOF mass spectrometry for bacterial identification, and the cefoxitin disc diffusion test for the screening of MRSA. Results: In total, the occurrence of MRSA colonization was 9.4% while Enterobacteriaceae colonization was 15.6%. No significant correlation was found for MRSA among departments or between day and night shifts. Alternatively, individuals working in the ICU and the operating room showed higher chances of being colonized by Enterobacteriaceae, and a notable connection was identified between Enterobacteriaceae and day shifts. Ultimately, the frequency of handwashing reduced the chances of being colonized by MRSA and Enterobacteriaceae, underscoring the significance of good hygiene practices. Conclusions: The study highlights the potential role of cleaning staff in transmitting HAIs, indicating a need for further research and consideration of enhanced hygiene protocols in hospital settings.

Background and Objectives: Catheter-associated urinary tract infections (CAUTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing and carbapenem-resistant Enterobacterales (CRE) significantly contribute to global rates of UTI. This study aimed to compare the prevalence and trends of ESBL-producing Enterobacterales and CRE in patients with CAUTIs and non-CAUTIs. Materials and Methods: A retrospective review of 4262 UTI-positive urine cultures was conducted at King Fahad Hospital of the University, Al Khobar, Saudi Arabia (January 2022–November 2023). Demographic, clinical, and microbiological data were obtained from hospital records. Antimicrobial susceptibility was tested using the Vitek^® System; ESBL and CRE were identified using Ezy MIC™ strips and Xpert^® Carba-R assay, respectively. Results: ESBL-producing Enterobacterales accounted for 11.3% of cases; CRE comprised 1.8%. ESBL was significantly more prevalent in non-catheterized patients and those in emergency care. CRE was significantly associated with catheterized patients and inpatient settings. Escherichia coli and Klebsiella pneumoniae were the predominant ESBL-producing and CRE isolates, respectively. bla-OXA-48 was the most frequently detected carbapenemase gene (66.7%). ESBL was prevalent in younger, non-catheterized females, suggesting increasing community transmission. Conversely, CRE were primarily observed in older, catheterized inpatients, emphasizing the role of invasive devices in resistance spread. Conclusions: These findings highlight the importance of targeted infection control and early catheter removal to mitigate resistance trends.