In this study, biofilm formation of Pseudomonas putida at a constant temperature of 5℃, 10℃, 20℃ or 30℃ under rich and poor nutrient conditions was investigated. For all temperature conditions, P. putida initially grew and formed biofilm. Subsequently, under the rich nutrient condition, the biofilm detached after it reached maturity at a high temperature, but those at a low temperature remained attached. In contrast, under the poor nutrient condition, biofilm detachment occurred regardless of the temperature condition; thus, lack of nutrients may cause biofilm detachment. Therefore, aside from cleaning blots and strict maintenance of a low temperature during the distribution of agricultural produce, attention must be paid to biofilm detachment as it can lead to an increased risk of bacterial contamination and food poisoning.