## 3 Results and Discussion
### Obtaining Unialgal Cultures  
Using some of the fresh samples, and before fixing them permanently, agar plates were inoculated and cultures were made in them to isolate the greatest possible number of microalgae species (Figure 11). The media used to isolate green algae were BBM (Phyto-Tech, Lenexa, KS, USA), while BG11 (Sigma-Aldrich, Madrid, Spain) media were used for cyanobacteria. From the beginning, the cultivation of diatoms was discarded, since they have greater nutritional demands and are a less common group. Finally, it was possible to isolate a total of 23 genera--10 green algae and 13 cyanobacteria (Table 3).  
The set of data is not completely representative of the site's microbiota, since it is known that the different species have different capacities for developing within in vitro cultures. Some species do not prosper in cultivation, even if they are predominant in the sample. Likewise, species that are not at all abundant in the samples may grow when cultivated, including species that are not observed in the sample, as has occurred with _Neochloris_ sp., _Pseudanabaen_ sp., some species of _Nostoc_ sp., _Gloecystis_ sp. and _Leptolyngbya_ sp. The latter genus grows very well when cultivated (Figure 9c,d), with up to 4 different strains having been isolated that are unidentified at the species level. This shows that there is a great diversity within this genus in the Alhambra and Generalife study areas. Moreover, in communities with great diversity it can be difficult to isolate different algae, especially filamentous cyanobacteria that form mats.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Fountains** & **Green Algae** & **Fountains** & **Cyanobacteria** \\ \hline
4 & _Neochloris_ sp. & 2 & _Dichothrix_ sp. \\
14 & _Stigeecolonium_ sp. & 4, 8, 15, 17 & _Leptolyngbya_ spp. \\
14, 8 & _Chlorococcus_ sp. & 20, 7 & _Clurococcus_ sp. \\
9, 12, 16, 19 & _Bractaococcus_ sp. & 2, 8 & _Pseudophormidium_ sp. \\
4, 8, 14 & _Scenedesmus_ spp. & 13 & _Pseudanabaena_ sp. \\
15 & _Pseudopleurococcus_ sp. & 6, 21 & _Calothrix_ sp. \\
5, 10 & _Klebsormidium_ sp. & 3 & _Pleurocarpsa_ sp. \\
11, 13 & _Choricyclis_ sp. & 3 & _Phormidium_ sp. \\
3 & _Apatococcus_ sp. & 5 & _Cyanosacrin_ sp. \\
4 & _Gloecystis_ sp. & 5 & _Schizol tririx_ sp. \\  & & 5 & _Nostoc_ sp. \\  & & 5 & _Chroococcidiopsis_ sp. \\  & & 4 & _Anmattoidea_ sp. \\ \hline \hline \end{tabular}
\end{table}
Table 3: Species obtained from unialgal cultures using samples collected in the Alhambra and Generalife. The fourtain numbers (Tables 1 and 2) from which the cultures came are indicated in the left column.  
Figure 11: Microalgal cultures: (**a**) an initial culture using a sample is shown on the left; (**b**) a unialgal culture of _Neochloris_ sp is shown on the right.  
It is useful to have isolated cultures of species, firstly for identification, since the microalgae's life cycles can be observed, as well as for other different characteristics that are often difficult to appreciate in samples. Furthermore, through this process a culture collection of living algae has been initiated that may be used in future studies. All of the microalgae from the Alhambra and Generalife are to a greater or lesser extent resistant to the treatments currently used to eliminate them (for example, many are resistant to chlorine), which makes them very interesting for use in trials with new treatments aiming to halt the proliferation of these organisms in research projects currently in progress.