The objective of this study was to evaluate the effects of antimicrobial drug use, gastric acid-suppressive agent use, and infection control practices on the incidence of Clostridium difficile-associated diarrhea (CDAD) in a 426-bed general teaching hospital in Northern Ireland. The study was retrospective and ecological in design. A multivariate autoregressive integrated moving average (time-series analysis) model was built to relate CDAD incidence with antibiotic use, gastric acid-suppressive agent use, and infection control practices within the hospital over a 5-year period (February 2002 to March 2007). The findings of this study showed that temporal variation in CDAD incidence followed temporal variations in expanded-spectrum cephalosporin use (average delay ؍ 2 months; variation of CDAD incidence ؍ 0.01/100 bed-days), broad-spectrum cephalosporin use (average delay ؍ 2 months; variation of CDAD incidence ؍ 0.02/100 bed-days), fluoroquinolone use (average delay ؍ 3 months; variation of CDAD incidence ؍ 0.004/100 bed-days), amoxicillin-clavulanic acid use (average delay ؍ 1 month; variation of CDAD incidence ؍ 0.002/100 bed-days), and macrolide use (average delay ؍ 5 months; variation of CDAD incidence ؍ 0.002/100 bed-days). Temporal relationships were also observed between CDAD incidence and use of histamine-2 receptor antagonists (H2RAs; average delay ؍ 1 month; variation of CDAD incidence ؍ 0.001/100 bed-days). The model explained 78% of the variance in the monthly incidence of CDAD. The findings of this study highlight a temporal relationship between certain classes of antibiotics, H2RAs, and CDAD incidence. The results of this research can help hospitals to set priorities for restricting the use of specific antibiotic classes, based on the size-effect of each class and the delay necessary to observe an effect.Clostridium difficile, a spore-forming gram-positive anaerobic bacillus, is a common pathogen in hospitals, with gastrointestinal colonization giving rise to increased morbidity, mortality, and health care costs (32). The clinical spectrum of Clostridium difficile-associated diarrhea (CDAD) ranges from uncomplicated diarrhea to severe life-threatening pseudomembranous colitis (23). Established risk factors for CDAD include host factors (for example, advanced age and comorbidities [21,28]), poor infection control practices (relating to the health care environment, health care workers' hand hygiene, etc.) (4, 21), exposure to factors that disrupt the normal protective intestinal microflora (i.e., broad-spectrum antibiotics) (6,24,25), and the use of gastric acid-suppressive agents, i.e., proton pump inhibitors (PPIs) (1, 8, 9) and histamine-2 receptor antagonists (H2RAs) (10). In a systematic review, which was undertaken to summarize the strength of the evidence for a relationship between antibiotic use and the occurrence of CDAD, most studies cited were limited in their ability to establish a causal relationship due to the presence of bias, small sample sizes, and the inadequ...