The purpose of this exploratory study was to assess occupational profile, level of performance, and on-the-job training needs of nursing staff employed in all government primary health care centers in rural Crete, Greece. The translated, culturally adapted, and validated Greek version of the Training Needs Assessment questionnaire was used. There were no significant differences between 2-year degree graduates (LPNs) and 3- or 4-year degree graduates (RNs, midwives, and health visitors) in terms of importance for 28 of 30 assigned tasks, whereas level of performance did not differ in any tasks. Significant training needs were reported by all staff, mainly in research/audit and clinical skills. Systematic overview of skill deficits in relation to skill requirements should be implemented by regional health authorities to enhance delivery of on-the-job training targeting group-specific, local needs.