Background: The goal of this study was to explore the effect of a shift work at adult trauma intensive care units on microbial contamination of cellular phones. Prevalence and type of microorganisms on intensive care unit (ICU) staff mobile phones is considered as a major route for transmission of infection. Methods: As a cross sectional study, cultures of mobile phone of 80 staff working at six adult trauma ICUs (54 beds) of a tertiary university affiliated hospital in Shiraz (Iran), were obtained, upon start and at the end of a shift. Samples were sent to the microbiologic laboratory of the hospital and cultured on specified media. Results: From 80 cell phones, 74 (92.5%) were contaminated upon entrance to ICU. Microorganisms included Staphylococcus coagulase negative 48 (60%), Bacillus 16 (20%), Acinetobacter 3 (3.75%) and others (Enterobacter, non hemolytic Streptococcus, E. coli and Serratia) with prevalence of 7 (8.75%). In comparison, exit cultures were positive for 49 (62.6%) of the cell phones that compromised of 34 (43.8%) coagulase negative Staphylococcus, 8 (10%) Bacillus, 2 (2.5%) Acinetobacter, and 5 (5.3%) contaminations with other microorganisms. Results showed a significant reduction in positive cultures at exit time from ICU. No correlation could be found between age, marital status, work experience in either hospital or ICU and changes in contamination of mobile phones. There was only a relationship between shift time and microbial colonization, which was more prevalent in night shifts followed by afternoon and morning shifts. Acinetobacter contamination of mobile phones of healthcare workers was more prevalent upon entrance to ICU than at exit time. Conclusions: High prevalence of cellular phone contamination with potential microorganisms is a major concern in adult trauma ICUs. Contamination of this equipment is more prevalent upon start of shift work, so a reverse transmission of bacteria may take place. Prevalence of Acinetobacter on entrance cultures necessitates special consideration. Infection control strategies to reduce cell phone colonization of trauma ICU staff, should be applied.