Morphological identification, isolation, and culturing of cyanobacteria derived from hot spring of Cisolok and Galunggung Mountain based on enrichment method

Abstract

Research on isolating and culturing of cyanobacteria from hot springs of Cisolok and Galunggung Mountain in West Java Indonesia had been done. This research was a study to find a fast and accurate method to identify, isolate and culture the cyanobacteria originating from extreme region, especially from the hot springs. Identification based on morphological characters performed in line with the process of isolation of the founded cyanobacteria strains. In the process of isolation, enrichment methods carried out in stages to determine the viability of each strain to be easily cultured (culturable). Four (4) types of media used in this study, i.e. BBM, BG-11, CT and MA. Pipette method was used to create a single culture. At the initial observations of the fresh samples, 3 types of cyanobacteria thallus were found, i.e. the form of thread/filament (unbranched filament), branched filament, and rounded shapes (coccoid). Furthermore, from all the enrichment samples (stage-1 and stage-2), 12 genera of cyanobacteria were found, i.e. Leptolyngbya, Gloeocapsa, Chroococcus, Oscillatoria, Synechococcus, Fischerella, Scytonema, Hapalosiphon, Microcystis, Nostoc, Gloeotrichia, and Stanieria. In the first stage of enrichment culture samples of Cisolok, eight (8) genera were found, i.e. Leptolyngbya, Chroococcus, Oscillatoria, Synechococcus, Scytonema, Hapalosiphon, Nostoc, and Gloeotrichia, while in the hot spring of Galunggung mountain, only four (4) genera were found (Leptolyngbya, Oscillatoria, Hapalosiphon, and Nostoc). There were several filaments and coccoid cyanobacteria that could not be identified by morphological characters. Meanwhile, in the second stage of enrichment culture samples of Cisolok, nine (9) genera were found, i.e. six (6) genera similar to those found in the first stage of enrichment (Leptolyngbya, Chroococcus, Oscillatoria, Synechococcus, Scytonema, Hapalosiphon), and added with the genus of Gloeocapsa, Microcystis, and Stanieria. From the hot spring of Mt Galunggung, nine (9) genera were found, i.e. four (4) genera similar to those found in the first stage of enrichment (Leptolyngbya, Oscillatoria, Hapalosiphon, and Nostoc), and added with the genus of Chroococcus, Gloeocapsa, Microcystis, Synechococcus, and Fischerella. Both from Cisolok and from Galunggung, the three genera (Leptolyngbya, Oscillatoria, Hapalosiphon) could be found in fresh samples and enrichment samples (stage-1 and stage-2). The research concluded that the enrichment process in stages (first and second) can be used for the isolation of strains of cyanobacteria to be adaptable, and therefore easy to be cultured (culturable). The genus Scytonema and Gloeotrichia are genera that known to be difficult to be cultured, and in this study, they grow only in stage-1, while the genus Stanieria appears in stage-2. Isolates obtained consist of 31 strains from Cisolok hot spring, and 13 strains from Galunggung mountain hot spring, then would be identified (re-identification) using molecular characters (16S rDNA).