## 3 Results and Discussion
### Demonstration-Scale (140 L)  
Regarding the environmental assessment at the demonstration scale, the characterisation results of the _Cyanobium_ sp. bioprocess are shown in Table 5 for the batch, and one litre of culture, with the respective mass and economic allocation for each extract. The relative contribution of each subsystem is depicted in Figure 4.  
As shown in Figure 4, the relative contribution was again relatively constant in almost all impact categories assessed, with the average contribution of S1 (11-17%), S2 (42-56%), S3 (0-1%), S4 (13%), S5 (5-7%), S6 (11-13%) and S7 (3-5%), with the cultivation phase (S2) having the greatest impact. The exception was observed in the ME impact category, \begin{table}
\begin{tabular}{c c c|c c c c c} \hline \hline \multirow{2}{*}{**Impact 1**} & \multirow{2}{*}{**Unit**} & \multirow{2}{*}{**Per Batch**} & \multirow{2}{*}{**Per Lite of Culture**} & \multicolumn{3}{c}{**Carotenoids**} & \multicolumn{3}{c}{**Phycobiliproteins**} \\ \cline{3-8}  & & & & **MA** & **EA** & **MA** & **EA** \\ \hline GW & kg CO2\({}_{\text{eq}}\) & 165.26 & 1.18 & 0.61 & 0.21 & 0.57 & 0.97 \\ SOD & kg CFC11\({}_{\text{eq}}\) & \(7.18\times 10^{-5}\) & \(5.13\times 10^{-7}\) & \(2.66\times 10^{-7}\) & \(9.29\times 10^{-8}\) & \(2.46\times 10^{-7}\) & \(4.20\times 10^{-7}\) \\ TA & kg SO2\({}_{\text{eq}}\) & \(7.25\times 10^{-1}\) & \(5.18\times 10^{-3}\) & \(2.69\times 10^{-3}\) & \(9.39\times 10^{-4}\) & \(2.49\times 10^{-3}\) & \(4.24\times 10^{-3}\) \\ FE & kg P\({}_{\text{eq}}\) & \(5.41\times 10^{-2}\) & \(3.86\times 10^{-4}\) & \(2.00\times 10^{-4}\) & \(7.00\times 10^{-5}\) & \(1.86\times 10^{-4}\) & \(3.16\times 10^{-4}\) \\ ME & kg N\({}_{\text{eq}}\) & \(7.19\times 10^{-3}\) & \(5.14\times 10^{-5}\) & \(2.67\times 10^{-5}\) & \(9.32\times 10^{-6}\) & \(2.47\times 10^{-5}\) & \(4.21\times 10^{-5}\) \\ TET & kg 1.4-DCB & 151.75 & 1.08 & 0.56 & 0.20 & 0.52 & 0.89 \\ FET & kg 1.4-DCB & 1.58 & \(1.13\times 10^{-2}\) & \(5.86\times 10^{-3}\) & \(2.04\times 10^{-3}\) & \(5.42\times 10^{-3}\) & \(9.23\times 10^{-3}\) \\ MET & kg 1.4-DCB & 2.25 & \(1.61\times 10^{-2}\) & \(8.36\times 10^{-3}\) & \(2.92\times 10^{-3}\) & \(7.74\times 10^{-3}\) & \(1.31\times 10^{-2}\) \\ HCT & kg 1.4-DCB & 3.37 & \(2.41\times 10^{-2}\) & \(1.25\times 10^{-2}\) & \(4.37\times 10^{-3}\) & \(1.16\times 10^{-2}\) & \(1.97\times 10^{-2}\) \\ HNCT & kg 1.4-DCB & 85.81 & 0.61 & 0.32 & 0.11 & 0.29 & 0.50 \\ FRS & kg oil\({}_{\text{eq}}\) & 49.37 & 0.35 & 0.18 & 0.06 & 0.17 & 0.29 \\ \hline \hline \end{tabular}  
* Impact category: global warming (GW), stratospheric ozone depletion (SOD), terrestrial acidification (TA), freshwater eutrophication (FE), marine eutrophication (ME), terrestrial ecotoxicity (FET), freshwater ecotoxicity (FET), marine ecotoxicity (MET), human carcinogenic toxicity (HCT), human non-carcinogenic toxicity (HNCT), fossil resource scarcity (FRS).  
\end{table} 
Table 5: Impact assessment results associated with the demonstration-scale process (140 L) per batch and per litre of culture, and the respective allocation for each co-product (mass allocation (MA) and economic allocation (EA)). where pre-inoculum accounts for 47% of the total impact, while cultivation only represents 23%, which can be explained by a more detailed analysis of the sub-systems. The relative contribution of S1 showed that the impact is mainly due to the reactor cleaning stage (ca. 70-80%), as it requires the use of significant amounts of sodium hypochlorite (responsible for 99% of this impact) and leads to the production of a large amount of wastewater which must be sent for further treatment. In addition, it should be noted that electricity and heat (for sterilisation) are still responsible for a large part of the impacts in the cultivation subsystem (S2): approximately 70% for electricity and 20% for thermal sterilisation.