[Isolation of atmospheric-nitrogen-fixing spirilla from the waters of the Paraná delta and other rivers].

Seventeen strains of Spirillum-like organisms (2) were isolated from the Parana Delta and other rivers (Table 1), using the following medium: 0.5% malic acid; 0.4% KOH; 0.5% K2HPO4; 0.005% yeast extract; pH was adjusted to 7 with KOH and 0.15% agar. The organisms produced a white, dense and sub-superficial pellicle in this medium, and streaking them on plates, single colonies could be isolated. They were easily recognized thanks to the Congo red added to the medium, because the bacteria, as it happens with Azospirillum species (6) (Spirillum lipoferum) (1), concentrated the strain. Using a colorimetric determination for ethylene (5), nitrogenase activity was detected in all the strains and in the NFb and potato-agar media the colonies were typical of Azospirillum (3). In PSS-semisolid and solid media, the growth was similar to that observed with Azospirillum strains. The isolated organisms developed in this medium better at 28 degrees C than at 37 degrees C, and the same was observed with 8 strains of Azospirillum isolated from plan roots of different species obtained from temperate regions. Some physiological tests were performed to identify the isolated organisms (Table 2). None of the isolated strains could be classified as a known species of Spirillum (4, 7), according to these tests. As thirteen strains showed properties in common with the terrestrial species A. brasilense and the others with A. lipoferum (8) they were tentatively identified as members of these species. Further studies are needed to ensure the classification of these strains and to determine the importance of these bacteria in the nitrogen balance of the waters from where they were isolated.