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Revista Colombiana de Biotecnología
Print version ISSN 0123-3475
Abstract
ORTIZ, Tatiana; OCAMPO, Valeria; PRADA, Luis Daniel and FRANCO-CORREA, Marcela. Preservation methods for actinobacterias with phosphate solubilizing activity. Rev. colomb. biotecnol [online]. 2016, vol.18, n.2, pp.32-39. ISSN 0123-3475. https://doi.org/10.15446/rev.colomb.biote.v18n2.47683.
The aim of this research was to evaluate different methods of preservation for actinobacteria with phosphate solubilization activity due to there are a few specific methods reported for these organisms. The methods were evaluated at three different time periods; long, medium and short-term and employing methods as cryopreservation and dry-freezing; clay, silica and sand; and periodically plating, respectively. Therefore 15 isolates from 3 different locations (La Vega, Maní and Tota) and a bank reference from Livestock Research Unit of the Pontificia Universidad Javeriana were used. All inocula were prepared in 0.85% saline solution (w/v) which were adjusted to a concentration of 108 cells/mL, followed each vial was inoculated with their respective storage cryoprotectants , glycerol 20% (v/v), 30% (v/v) to freeze and 18% skim milk (w/v) for dry-freezing. Medium-term methods were performed similarly; the inoculum was added to 10 clay beads, 10 g of sand and 5 g of silica and then stored at 4 °C. The short-term method was evaluated in oatmeal agar (15 g/L). The evaluation was performed by direct counting in a Neubauer chamberusing trypan blue staining technique, in addition to macroscopic and microscopic characterization of each isolate in periodic plating. It was established that the phosphorus solubilizing activity was more stable in glycerol 30% (w/v) and lyophilization for statistical analysis.
Keywords : Viability; cryoprotectants; bank; dry-freezing; cryopreservation.