Many organizations have built efforts towards fostering diversity and inclusion within agricultural education. While many efforts focus on programming, no research was found focusing on root issues such as prejudices held by teachers. Thus, the purpose of this study was to investigate levels of prejudice, as defined by integrated threat theory, held by secondary agriculture teachers. A survey addressing social desirability, sexism, racism, and homophobia was distributed to a random sample of Agricultural Education teachers. After controlling for nonresponse error, response was 37.5% (n = 135). Social desirability (α = .6) did not correlate with any prejudice variables, indicating results weren’t biased in this way. All other scales (α > .8) indicated moderate prejudice toward women (M = 2.09, SD = .57) and lesbians (M = 2.14, SD = .89); and slight prejudice toward gay men (M = 1.93, SD = .88) and racial minorities (M = 1.93, SD = .53). A Multivariate Analysis of Variance (MANOVA) was ran and results indicated significant results when comparing prejudice levels in both gender, F(4, 101) = 2.997, p = .022, Λ = .894, η2p = .11; and population of hometown, F(4, 101) = 3.201, p = .016, Λ = .887, η2p = .11. However, there was no significant interaction of the two factors, F(4, 101) = 0.643, p = .633, Λ = .975. Univariate test results show that male participants had higher levels of sexism, F(1, 104) = 7.896, p = .006, η2p = .07; and homophobia toward lesbians, F(1, 104) = 7.91, p = .006, η2p = .07; and gay men, F(1, 104) = 8.921, p = .004, η2p = .08. In addition, participants from rural hometowns showed higher levels of homophobia toward lesbians, F(1, 104) = 6.457, p = .013, η2p = .06; and gay men, F(1, 104) = 12.052, p = .001, η2p = .1; and racism, F(1, 104) = 4.817, p = .03, η2p = .04. This study can be useful in identifying potential roots of prejudices and we recommend that agricultural education state staff and teacher educators seek ways to integrate forms of tolerance within their programs.