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Staphylococcus spp. were present in all samples, and Micrococcus spp. were present in all of the extended semen samples although they were also isolated from the extender. The number of bacteria in colony forming units per mL varied considerably among samples. Only one microbe known to be associated with equine infertility, Pseudomonas spp., was isolated from three samples, albeit in low numbers. In conclusion, bacterial culture followed by MALDI-TOF does not identify all bacteria present in stallion semen samples. In-depth knowledge of