The effect of extensive urban dust pollution, caused mainly by road traffic, on some biochemical and structural characteristics of current-year Norway spruce (Picea abies L. Karst.) needles was investigated. Two categories of needle samples were formed according to the data about the pollution levels obtained from the Croatian National Institute of Public Health: less and more affected. Apoplastic guaiacol peroxidases were used as the molecular stress markers. Peroxidase activity was doubled in more affected needles compared with the less affected ones. Also, the electrophoretic pattern of samples extracted from more affected needles revealed the expression of additional isozyme band, which could be attributed to the activation of detoxifying mechanisms. Anatomy of more affected needles was changed as well. Necrosis of needle mesophyll usually connected with the stomata was the most outstanding character. Also, distortions of sieve cells were present in the same needle samples indicating possible disturbances in mineral nutrition. The obtained results showed that needles of Norway spruce trees that are exposed to the higher pollution level undergoes to both structural and biochemical changes. Besides of the described changes, the investigated spruce trees are able to survive in more polluted environment as well.