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Healthcare staff is one of the professional groups that suffers the highest exposure to sources of occupational stress such as hostility from coworkers and superiors. In order to contribute to the assessment of bullying behaviors in the healthcare sector and to obtain a brief and manageable instrument for the assessment of this psychosocial risk, we developed the Hospital Aggressive Behaviour Scale-version Co-workers-Superiors (HABS-CS). By means of thorough qualitative analysis, an initial pool of 166 items was obtained, which were reviewed according to precise criteria until concluding with a total of 57 items, which were administered to a sample of 1,484 healthcare professionals from 11 public hospitals. The analyses concluded with the selection of 17 items distributed in two subscales. The internal 5-factor structure is the result of exploratory and confirmatory factor analysis conducted in two samples. Both the resulting questionnaire and the factors identified present adequate psychometric properties: high-internal consistency (Cronbach's α of .86) and adequate criterion validity, analyzed by means of significant correlations between the HABS-CS and job satisfaction, burnout components, and psychological well-being. This instrument may be of great utility for the assessment and prevention of psychosocial risks.