Clostridium spp. are ubiquitous bacteria and often found in foods and animals. Some species are pathogenic, others food spoiling or commensals. In this study, 65 cold-tolerant Clostridium spp. strains isolated from variable samples (beef, lamb, venison, feces/skin of wild boars) were investigated. Fifty strains were lecithinase positive; six additionally produced β-hemolysis. By applying specific qPCR, 16S rRNA gene analysis, RFLP method, and MALDI-TOF MS, they were classified into two major groups: 29 strains were identified as C. tagluense-like, while the other 36 remained unidentified. Subsequently, twenty-two vacuum-packed beef samples were spiked with a single strain from both groups and stored at 4 °C for 8 weeks. The odor of challenged samples was variable (from unchanged, sour/musty, to sulfurous), while color, meat consistency and drip loss were similar to the control group. The ability to produce gas of all tested strains was lower than of C. estertheticum. Even though both groups of cold-tolerant clostridia exhibited similar 16S rRNA genes and biochemical activities, RFLP methods and MALDI-TOF MS are sufficient to differentiate them. In terms of food safety, strains producing lecithinase and hemolysin should be further investigated for their potential to produce substances affecting human and animal health.

梭菌属 是无处不在的细菌，经常在食物和动物中发现。有些物种是致病性的，其他物种会导致食物变质或共生。在这项研究中，65 种耐寒梭菌属。研究了从可变样品（牛肉、羊肉、鹿肉、野猪的粪便/皮肤）中分离的菌株。50 株卵磷脂酶阳性；6另外产生β-溶血。通过应用特异性qPCR、16S rRNA基因分析、RFLP方法和MALDI-TOF MS，将它们分为两大类：29株被鉴定为C. tagluense-like，而其他36个仍然身份不明。随后，在 22 个真空包装的牛肉样品中加入来自两组的单一菌株，并在 4°C 下储存 8 周。挑战样品的气味是可变的（从不变、酸味/霉味到硫磺味），而颜色、肉稠度和滴水损失与对照组相似。所有受试菌株产气能力均低于C.estertheticum。尽管两组耐寒梭菌表现出相似的 16S rRNA 基因和生化活性，但 RFLP 方法和 MALDI-TOF MS 足以区分它们。在食品安全方面，应进一步研究产生卵磷脂酶和溶血素的菌株产生影响人类和动物健康的物质的潜力。