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Background: Ventilator‐associated pneumonia is common and associated with high mortality. Nurses play a fundamental role in preventing ventilator‐associated pneumonia. Evidence‐based guidelines and training interventions about preventing ventilator‐associated pneumonia do not ensure compliance with recommendations. Aims and objective: To evaluate the impact of nursing workload on compliance with non‐pharmacological measures to prevent ventilator‐associated pneumonia. Design A prospective observational study in two medical‐surgical adult ICUs in Spain. Methods: We measured nurses' knowledge about preventing ventilator‐associated pneumonia through a questionnaire and workload, categorized with the Nine Equivalents of Nursing Manpower Use Score. We directly observed nurses to measure compliance with non‐pharmacological measures to prevent ventilator‐associated pneumonia. Results: A total of 97 nurses were studied; 76 (79%) were observed. There were 327 observations (mean 4·30 observations per nurse). The questionnaire showed good baseline knowledge of preventive measures [mean score (92% ± 16%)]. Nurses complied with the preventive measures in 66% ± 15% of observations; compliance ranged from 11·9% for pre‐aspiration hand washing to 99·7% for using sterile aspiration probes. Mean Nine Equivalents of Nursing Manpower Use Score for each nurse observed was 50 ± 13, without differences between centres (49 ± 14 versus 51 ± 11, p = 0·4). Overall compliance was lower in the light workload group (p = 0·02), but no significant differences in compliance between workload groups were found when each measure was analysed separately. Compliance was higher in nurses aged 31–40 years than in those aged >51 years, although workload was similar in both groups. Conclusions: Compliance with non‐pharmacological measures to prevent ventilator‐associated pneumonia varies widely; non‐compliance is not because of increased workload. Relevance to clinical practice: Most critical care nurses have good baseline knowledge of non‐pharmacological measures to prevent ventilator‐associated pneumonia. Failure to comply with these measures is probably more related with behavioural, structural and organizational aspects than with nursing workload. Interventions to improve compliance might be more effective if they focus on factors such as work climate and professionals' attitudes. [ABSTRACT FROM AUTHOR]