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Physical stability and survival of microorganisms stored in water-in-oil emulsions

In water-in-oil (w/o) emulsions containing microorganisms, Laplace pressure driven water 
transport from small cell-free droplets to large cell-associated droplets reduces the physical stability 
of the emulsions and biological stability of microorganisms. To prevent water transport, we 
investigated adding the solute NaCl to the aqueous phase of the emulsion. Forming a mixed 
emulsion where one phase contained NaCl and the other contained cells was imperative for 
minimizing solute associated fungitoxicity. When the mosquito control agent Lagenidium giganteum 
was formulated in an emulsion simultaneously with a 5 wt% NaCl solution, its viability and control 
efficacy were significantly lower compared to the same ingredients prepared as a mixed emulsion 
indicating the oil acts as a barrier to solute transport between aqueous droplets. The death rate for 
cells stored in a mixed w/o emulsion with salt in a separate phase was 10% lower compared to 
storage in a w/o emulsion without salt during the first 30 days of storage indicating the presence of 
salt may improve the short-term storage of cells. Particle size distribution measurements confirmed 
water transport in emulsions consistent with Ostwald ripening. The presence of solute in the aqueous 
phase reduced droplet size increases in cell-associated and cell-free droplets.