Smoking and Clostridioides difficile Colonization in Adults Aged ≥40 Years: Findings from a Community-Based Gut Microbiota Study

Hong-Ming Chen,1,2,* Te-Wei Yang,3,* Wen-Chien Ko,3,4 Yuan-Pin Hung,5,6 Chin-Shiang Tsai,3,4 Pei-Jane Tsai7 1Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan; 2Yanshuei District Health Station, Tainan, Taiwan; 3Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan; 4Center for Infection Control, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan; 5Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan; 6Department of Internal Medicine, Tainan Hospital, Ministry of Health and Welfare, Tainan, Taiwan; 7Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan*These authors contributed equally to this workCorrespondence: Pei-Jane Tsai, Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, Email [email protected]: Asymptomatic Clostridioides difficile colonization (CdC) serves as a reservoir for pathogen transmission and may precede clinical infection. While risk factors for CdC have been well described in hospitalized populations, community-based data—particularly regarding lifestyle-associated factors and gut microbiota alterations—remain limited in Taiwan. This study aimed to determine the prevalence and risk factors of community-acquired CdC and to explore associated gut microbiota differences.Materials and Methods: We conducted a cross-sectional study analyzing 250 residual stool samples from adults aged ≥ 40 years who participated in community health screenings in Tainan City between 2006 and 2009. CdC was detected by polymerase chain reaction targeting the triosephosphate isomerase (tpi) gene. Demographic characteristics, lifestyle factors, and laboratory parameters were collected and analyzed. Exploratory 16S rRNA gene sequencing was performed on selected samples to compare gut microbiota composition between CdC and non-CdC groups.Results: The prevalence of CdC was 11.6% (29/250). Cigarette smoking was identified as the sole independent factor associated with CdC (adjusted OR 2.35, 95% CI 1.05– 5.28; P = 0.038). Exploratory microbiota analysis revealed differences in community composition between CdC and non-CdC samples, including increased relative abundance of Proteobacteria in CdC.Conclusion: Cigarette smoking was associated with an increased likelihood of community-acquired CdC in southern Taiwan. Distinct gut microbiota profiles were observed in individuals with CdC, supporting a potential link between smoking-related microbiota alterations and susceptibility to asymptomatic colonization. These findings underscore the relevance of modifiable lifestyle factors in the epidemiology of community CdC and warrant further investigation.Keywords: Clostridioides difficile colonization, smoking, gut microbiota, community, Taiwan