Improving the hygienic status of feed ingredients by biotechnological processes as fermentation is of the greatest concern. This preliminary study aimed to investigate whether there are relevant effects of fermented liquid feed (FLF) on the survival of potential pathogens in vitro. The feed (fresh basis) consisted of 50% rye, 30% rapeseed extracted meal, 10% barley and 10% wheat. Glass bottles were filled about 14.1 g water (38 °C) containing the diluted starter culture and feed (8.81 g). Fermentation led to high levels of lactate (5−7% of dry matter), low pH values (<4.0) and low levels of acetic acid (<1% of dry matter) in the FLF. The survival rate of pathogens added, such as Salmonella enterica serovar Typhimurium, Escherichia coli and Clostridium perfringens after 6 h of controlled fermentation, was significantly reduced (<2 log10 CFU/g). The counts of Candida krusei in FLF at 3 h and 6 h post inoculation remained almost unchanged regardless of the incubation time. Even adding sodium-benzoate at a concentration of up to 0.25% in the liquid feed did not reduce the survival of C.krusei during fermentation. Based on this in vitro study, feeding of FLF seems a promising strategy to reduce pathogen transmission but has to be confirmed on natural feeds by pathogens for increasing the hygienic properties.