Clostridium perfringens is one of the most common causes of food poisoning in the world. In this study, C. perfringens isolated from raw milk, milk products and human consumers were investigated for their antibiotic resistance and biofilm formation ability. A total of 460 samples (buffalo milk, cow milk, camel milk, yoghurt, Kareish cheese and soft (processed) cheese, 60 of each and 100 diarrheic stool sample of human consumers) were collected from Zagazig city, Sharkia Governorate, Egypt, and examined for the presence of C. perfringens strains. Twenty-four C. perfringens strains were isolated and all were of type A and 3 (12.5%) of camel milk origin and 4 (16.7%) from human diarrheic stool were positive for the enterotoxin associated (cpe) gene. Antibiotic sensitivity revealed that 87.5% of the isolates were resistant to oxytetracycline followed by amoxicillin (83.4%), ampicillin and erythromycin (75%, each). C. perfringens isolates from different sources were able to form biofilm at various temperatures (4°C, 25°C and 35°C). In conclusions, the collected samples from the study area are considered potential sources for human infection with C. perfringens. Therefore, milk and dairy products should be inspected periodically to control contamination with foodborne pathogens.