Encapsulation in alginate-polymers improves stability and allows controlled release of the UFV-AREG1 bacteriophage

Abstract

The bacteriophage UFV-AREG1 was used as a model organism to evaluate the encapsulation via extrusion using different hydrocolloids. Pure alginate [0.75%, 1.0%, 1.5% and 2.0% (m/v)] and mixtures of alginate [0.75% or 1.0% (m/v)] with carrageenan [1.25% (m/v)], chitosan [0.5% (m/v)], or whey protein [1.5% (m/v)] were used to produce bacteriophage-loaded beads. The encapsulating solutions presented flow behavior of non-Newtonian pseudoplastic fluids and the concentration of hydrocolloid did not influence (p > 0.05) the morphology of the beads, except for alginate-chitosan solutions, which presented the higher flow consistency index (K) and the lower flow behavior index (n). The encapsulation efficiency was about 99% and the confocal photomicrography of the encapsulated bacteriophages labeled with fluorescein isothiocyanate showed homogenous distribution of the viral particles within the beads. The phages remained viable in the beads of alginate-whey protein even when submitted to pH 2.5 for 2 h. Beads incubated directly in simulated intestinal fluid (pH 6.8) resulted in a minimal of 50% release of the UFV-AREG1 phages after 5 min, even when previously submitted to the simulated gastric fluid (pH 2.5). Encapsulation enabled phages to remain viable under refrigeration for five months. Encapsulated UFV-AREG1 phages were sensitive to dehydration, suggesting the need for protective agents. In this study, for the first-time bacteriophages were encapsulated in alginate-carrageenan beads, as well as alginate-chitosan as a bead-forming hydrocolloid. In addition, a novel procedure for encapsulating bacteriophages in alginate-whey protein was proposed. The assembled system showed efficiency in the encapsulation of UFV-AREG1 bacteriophages using different hydrocolloids and has potential to be used for the entrapment of a variety of bioactive compounds.

Keywords: Carrageenan; Chitosan; Escherichia coli O157:H7; Phage therapy; Rheology; Whey protein.