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In the past decade, the number of couples consulting an assisted reproductive techniques (ART) center for infertility has increased in most European countries. In France, sperm bacterial examination must be performed every 6 months in couples undergoing in vitro fertilization (IVF) cycles, according to 2010 French ART Guidelines. The aim of this study was to analyze the results from such samples at Nantes University Hospital and to assess their potential value for infertile couples undergoing ART cycles. Between 2003 and 2008, semen cultures performed were analyzed according to bacterial enumeration, type and number of bacterial species, and their antibiotic resistance profile. A total of 14,119 semen cultures were performed, showing an annual increase of 45% from the start to the end of the study. The proportion of positive semen cultures was stable throughout the study period (40 to 45%). Many bacterial species were considered as contaminants (coagulase-negative staphylococci, alpha-hemolytic streptococci). For pathogen agents (in most cases, Enterobacteriaceae), the antibiotic resistance profile revealed mostly a susceptible phenotype. Finally, every positive bacterial result had direct consequences on the IVF cycle management, with subsequent reinforced advice on the hygiene procedure before sample collection and/or antibiotics prescription.