This study investigated the comparative influence of ethnicity and religious affiliation on the alienation of Nigerian university staff from their work environment. The influence of certain moderator variables such as the location of the university, gender, age, educational qualification, staff category, official rank and staff communicative ability in the dominant Nigerian language spoken on campus were also investigated. The three instruments used in the collection of data were Ethnic Affiliation Scale (Cronbach alpha coefficient 0.945), Religious Affiliation Scale (Cronbach alpha coefficient 0.887) and Staff Alienation Questionnaire (Cronbach alpha coefficient 0.840). The total sample used was 532 members of both teaching and non-teaching staff selected by stratified sampling from six Nigerian universities purposefully chosen. Stepwise multiple regression was applied in the analysis and the critical level of significance applied in all the analysis was never less stringent than 0.05. The major findings were as follows. Ethnicity and religious affiliation variables predicted staff alienation from co-workers, but religious affiliation variables alone predicted staff alienation from friends at work and from the job. Religious affiliation was found to be stronger than ethnicity in predicting the alienation of university staff from their co-workers (p less than 0.001), their friends at work (p less than 0.001) and their jobs (p less than 0.001). Staff of north-located Nigerian universities felt more alienated from their co-workers and their jobs than those in the east and west. But university staff in the north were least alienated from their friends at work. Staff of universities located in the west felt most alienated from their friends at work. (Contains 3 tables.)