Hydrogels based on poly(vinyl alcohol), PVA, and calcium alginate were prepared by a freezing and thawing cycle process and characterized, in terms of the role of the polymer mixture percentage and the number of treatment cycles, on their weight swelling ratio, WSR, gel fraction, and activated sludge entrapment and immobilization. The results show that the morphology of these hydrogels is highly dependent on the PVA-Ca alginate ratio of 5 wt % total polymer content in the initial aqueous solution and that the number of entrapped microorganisms which survive the freezing-thawing procedure is independent of this ratio. For 80/20 PVA-Ca alginate hydrogels, results also show that for up to three freezing and thawing cycles, the WSR, which is in average 24, is not severely affected by the number of the cycles. For the hydrogels with three cycles, the calculated gel fraction for the composite hydrogel is 0.99. Immobilized microorganisms from sedimented activated sludge, constituted by bacteria and fungi, die in high numbers during the freezing and thawing treatment. However, with a proper time of incubation with glucose as carbon source, the population of bacteria is recovered and mainly proliferate inside the hydrogel, attached on top of the fibril network formed by the polymers, while fungi are recovered predominantly on the surface of the spheres.