Medical mistrust among racial/ethnic minorities has been associated with decreases in health care utilization, whereas religiosity has been separately linked with increases in this behavior. However, very few studies have examined the relationship between religiosity and medical mistrust among Latinos, a group with strong religious connections and potentially high mistrust. In-person, self-administered surveys were collected among 767 adult Latinos attending three Latino churches (one Catholic and two Pentecostal) in Long Beach, CA. Measures included a previously validated 12-item medical mistrust scale, religiosity (religious denomination, length and frequency of attendance, and number of groups or ministries involved in), health care access, and sociodemographic factors. Medical mistrust score was 2.47 (standard deviation [SD] = 0.77; range 1-5). Almost two-thirds of participants (62%) attended religious services frequently (once a week or more), and the majority attended a Catholic church (80%). About half of the participants had attended their church for ≥5 years (50%) and participated in one to two church groups or ministries (53%). Multivariable analyses show that Pentecostal church congregation and those identifying as Mexican/Chicano were negatively associated with medical mistrust. On the contrary, participating in church groups or ministries and having an immigrant parent were positively associated with medical mistrust. Our findings suggest that church-based health initiatives should consider church denomination, length of attendance, participation in groups or ministries, and ethnic differences to address medical mistrust issues among Latino congregants.