Polymers selection for a liquid inoculant of Azospirillum brasilense based on the Arrhenius thermodynamic model

Abstract

Plant growth promoting bacteria (PGPB) enhances the growth of their hosts and can protect them from biotic and abiotic stresses. Bacterial inoculants contain one or more of these beneficial strains in a carrier material, which must be able to maintain the viability of the cells during the time of storage, and also guarantee the biological activity of the strains once applied in the soil. These inoculants can be solid, liquid, gel or oil-based, depending on the characteristics of the strains and the shelf life expected by the producers. In this study, we used a method of accelerated degradation to select a polymer and a concentration to maintain cell stability of a liquid inoculant based on the strain C16 Azospirillum brasilense. A screening at 45°C was made to compare the protectant effect of five polymers on the viability of the strain (p/v): carrageenan (1.5%), sodium alginate (1%), trehalose (10 mM), polyvinylpyrrolidone (2%), glycerol (10 mM) and phosphate saline buffer as control. Carrageenan and sodium alginate showed significant differences in cell viability over the use of other polymers (P < 0.05). We evaluated cell viability with these two polymers at three concentrations and three different temperatures (4, 28 and 45°C) for 60 days and determined the bacterial degradation rates. Based on the Arrhenius thermodynamic model, we calculated the time required to reduce cell concentration in three log units, and observed that the protectant activity of each polymer and each concentration depends on the temperature of storage. Cell viability was best preserved in all treatments at 4°C. In general, alginate prolonged cell viability at 28°C, and carrageenan at 45°C. Alginate at 1% and  carrageenan at 0.75% showed a stable behavior (superior to the control) in the three evaluated temperatures, so we conclude that they can be used for a  formulation of a liquid inoculant based on the strain C16 of A. brasilense.

Key words: Energy of activation, degradation, cell death, kinetics, formulation.