Spondylodiscitis is a severe spinal infection that requires an effective antibiotic treatment. Therefore, we sought to analyse the causative pathogens from intraoperative specimen in patients with spondylodiscitis and a need for surgery. To this end, we performed a retrospective study of all patients with spondylodiscitis and a need for operative treatment admitted to our hospital between January 2020 and December 2022. A total of 114 cases with spondylodiscitis were identified. A total of 120 different pathogens were detected. Overall, 76.7% of those microorganisms were Gram-positive bacteria. The most common causative pathogen was Staphylococcus aureus (n = 32; 26.6%), followed by coagulase-negative staphylococci (n = 28; 23.4%), of which Staphylococcus epidermidis (n = 18; 15%) was the most frequently detected, as well as enterococci (n = 10; 8.4%) and Streptococcus spp. (n = 11; 9.2%). Moreover, 19.1% (n = 22) and 3.4% (n = 4) of all detected isolates were Gram-negative pathogens or fungi, respectively. Overall, 42.8% of all coagulase-negative staphylococci were oxacillin-resistant, while none of them were vancomycin-resistant. In summary, 50% of the pathogens could be identified as staphylococci. The results of our study highlight the important burden of oxacillin-resistant Gram-positive bacteria as an aetiological cause of spondylodiscitis, providing a relevant finding for antimicrobial stewardship programmes.