Anaerobic bacteria are of great concern to the food industry, especially Clostridium perfringens that produce exotoxins. The present study was performed on 67 liver samples from aged female buffaloes (over 6 years old) slaughtered in the slaughter houses in Assiut Governorate, Egypt. Forty-four liver samples showed gross hepatic lesions and twenty-three samples were apparently normal liver. Bacteriological examination of these liver samples revealed that 48 samples (71.64%) were positive for anaerobic bacterial isolation, in mixed infection 41.7% and in single infection 58.3%. All Fasciola infected livers were positive for anaerobic bacterial isolation. Clostridium perfringens was isolated in mixed form with incidence 29.2%, 14 out of 48 samples positive for anaerobic culture, (25% and 4.2% from liver samples showed hepatic affections and normal livers, respectively). The Pathogenicity of the isolated Clostridium perfringens in mice revealed that 64.3% and 35.7% were toxigenic and non-toxigenic, respectively. The most prevalent anaerobic isolates were Bacteroides spp. and Peptostreptococcus anaerobius either in single or mixed form. In addition Fusobacterium necrophorum was isolated from two liver abscesses in a single form (4.2%). Antimicrobial susceptibility testing of the most prevalent isolates, with 12 antibiotics, revealed their sensitivity 100% to Ofloxacin and Ciprofloxacin followed by Oxytetracycline (94.1%) and Erythromycin (76.5%). All tested bacterial isolates showed resistance to Colistin, Amoxicillin, Spectinomycin and Rifamycin. It can be concluded that livers of slaughtered aged female buffaloes considered as hazardous source of mixed     anaerobic bacteria especially which are infected with Fasciola.