Bacterial diversity of biofilms on polyhydroxybutyrate exposed to marine conditions: Ex-situ vs. in-situ tests

Biofilms form on any available surface and, depending on the characteristics of the material and the environmental conditions, biodegradation can take place. We compared the bacterial composition of polyhydroxybutyrate (PHB)-related biofilm communities from marine ex-situ and in-situ tests to assess the differences in diversity and abundance between these two biofilms. This comparison will help to better assess the transferability of tank tests to real-life scenarios. The in-situ tests were set up in the Mediterranean Sea on the Island of Elba, Italy where PHB-tensile bars were lodged in the sediments. This created a water-exposed aerobic and mud-planted anaerobic scenario. The ex-situ tests were modeled after in-situ tests and performed in temperature-controlled tanks. The PHB-related biofilms were harvested after 240 days of exposure along with planktonic bacteria, and particle- and sediment-related biofilm. The bacterial composition was elucidated using 16S rDNA sequencing. Biofilms harvested from the in-situ test were more diverse, less even, and contained more rare species compared to biofilms from the ex-situ test. The PHB-related biofilm was characterized by a higher abundance of the bacterial order Desulfobacterales. The composition of PHB-related biofilm varied significantly between the two tests and between aerobic and anaerobic conditions. The composition of PHB-related biofilm was significantly different from planktonic bacteria, particle, and sediment-related biofilm, showing the influence of PHB on the biofilm composition. Thus, the ex-situ tank test for PHB degradation cannot, in terms of bacterial composition, simulate the in-situ conditions to their full extent.