ABSTRACT Purpose: At Matsuyama University, all departments hold lectures regarding smoking prevention and cessation during freshman orientation. To examine changes in the amount of knowledge regarding the subjective harmful effects of smoking as well as in the attitude toward smoking before and after lectures, questionnaire surveys were conducted to assess the educational effects of such lectures. Methods: Among students who attended the Matsuyama University freshman orientation in 2016, we included 560 students who provided consent and responded to a questionnaire survey. Examination items included age, gender, presence or absence of a smoking habit, when the individual began smoking (smokers only), amount of knowledge regarding smoking (individual subjective evaluation), attitude toward smoking among non-smokers, and attitude toward smoking cessation among smokers. Changes in the amount of knowledge and attitude before and after the lectures were examined using a Wilcoxon signed-rank test, and an intergroup comparison of the amount of knowledge was performed using the Steel–Dwass test. Results: With smoking prevention and cessation education, 38 of 60 students who had responded that they “might smoke” before the lectures changed their attitude to “will not smoke.” No significant differences were observed in the amount of knowledge between them and 483 students who responded that they “will not smoke” before and after the lectures. Moreover, 22 students who did not change their attitude to “will not smoke” even after the lectures were markedly less mindful about smoking; however, the amount of knowledge these students possessed after the lectures was significantly lower than that possessed by the other group of students. On the other hand, students who were smokers showed no change in their attitude to quit smoking. Conclusion: Smoking prevention and cessation education during university admission was found to be useful. Moreover, continuous lectures with easy-to-understand content and increasing the amount of knowledge with regard to the harmful effects of smoking were shown to effectively raise awareness about smoking prevention and cessation.