To study the effect of Enterococcus spp. on C. botulinum strains, heat treated spores or vegetative cells were added to RCM medium at a final concentration of 104 cfu/ml. The inhibitory effect of different Enterococcus spp. was studied by addition of different bacterial dilution (101–109 cfu/ml to C. botulinum culture medium. The mixture was incubated anaerobically at 37 °C for 5 d. C. botulinum was quantified using the most probable number (MPN) estimation method using Differential Reinforced Clostridial broth (DRCM, Sifin, Germany) and the neurotoxins were tested using an ELISA