Street food vending is a common feature of most cities and towns in developing countries. Hygienic conditions are frequently not ideal, and may result in microbial contamination and foodborne diseases. Our research focused on evaluation of food safety knowledge and hygiene practice of street vendors in Mekong river delta region, Vietnam. Our results showed that only 54.4% of street vendors reaching the food safety condition. Meanwhile, the infrastructure with 67.9% and document with 73.1% were recorded. Most of street vendors had knowledge of food safety with 73.0%. Whereas, group of food selection and preservation ability acquired with 83.5%; group of general knowledge about food safety acquired with 55.8%. Regarding to handling, street vendors had hygiene practice with 98.5%, the maid had 98.5% and personal hygiene had the lowest percentage with 33.1%.