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The Eating and Food Literacy Behaviors Questionnaire (EFLBQ) is an instrument that has not been tested for its capacity to distinguish among individuals assumed to have higher and lower food literacy. The aim of this study was to examine and compare EFLBQ scores among university students with and without formal nutrition-related training. This study had a cross-sectional design. Two hundred twenty-seven young adult university students enrolled in non–nutrition-related classes (n = 76), introductory nutrition classes (n = 98), and senior-level nutrition classes (n = 53) were recruited from a large university in the southeastern United States during the spring and fall semesters of 2022. The students ranged in age from 18 to 30 years. Mean EFLBQ total scores and factor scores of health and nutrition, taste, food preparation, planning and decision making, and convenience were compared among the three groups. Linear regression backward method examined whether level of training, age, sex, race and ethnicity, and BMI were independent variables associated with total EFLBQ mean scores. Analysis of variance (ANOVA) compared normally distributed data and Kruskal-Wallis and Mann-Whitney U tests compared non-normally distributed data. Linear regression testing found that the level of nutrition training was significantly associated with total EFLBQ scores. Students in senior-level nutrition classes had higher total EFLBQ mean scores (M = 3.1 ± 0.2) than students in non–nutrition-related classes (M = 2.9 ± 0.3) (P = 0.002; 95% CI = 0.055, 0.280) and introductory nutrition classes (M = 2.9 ± 0.3) (P = 0.001; 95% CI = 0.275, 0.061). Total EFLBQ mean scores were not different among the students in non–nutritional-related classes and introductory nutrition classes (P = 1.000; 95% CI = −0.096, 0.096). Scores differed by training level for items measuring EFLBQ Factor 1: Health and Nutrition. Factor 1 mean scores were higher in students in senior-level classes (M = 3.1 ± 0.4) than for those in non-nutrition classes (M = 2.7 ± 0.5) (P < 0.001; 95% CI = 0.294, 0.670) and introductory nutrition classes (M = 2.8 ± 0.5) (P < 0.001; 95% CI = 0.228, 0.553). The other EFLBQ factor mean scores were not different among the three groups (all P -values > 0.12). The EFLBQ demonstrated the capacity to distinguish food literacy scores between students expected to have higher as compared with lower food literacy. [ABSTRACT FROM AUTHOR]