The two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae) is a major pest mite infesting vegetables and horticultural ornamental and agronomic crops in Egypt. In biological control strategies, natural enemies were applied to maintain TSSM population below the level where damage occurs for an extended period of time besides utilizing acaricides in a critical situation as a quick practice to avoid deleterious TSSM damage. This study evaluated the potential of acaricides (Abamectin, Chlorfenapyr, Fenpyroximate and Hexythiazox ) and the pathogenicity of native species of Trichoderma spp.(Trichoderma harzianum and T.album) on different biological developmental stages of females and males of T. urticae and Phytoseiulus persimilis Athias- Henriot and their population under greenhouse conditions. Results revealed that the utilization of recommended application rate (RC) of acaricides showed a deleterious effect on the different biological developmental stages including eggs, larvae, protonymh, deutonymph and adults of female and male of TSSM, T. urticae and predaceous mite, P. persimilis under laboratory conditions. Moreover, the effects varied greatly on immature stages, life cycle, adult stage, longevity and life span of TSSM and P. persimilis according to utilized acaricide. Concerning the pathogenicity of the two bio-agents (T.harzianum and T.album and a mixture of them), T.harzianum was more pathogenicity against T. urticae and P. persimilis than T.album. Moreover, Trichoderma species showed a negative effect on the population density of T. urticae than its predaceous mite in vitro treatments and under greenhouse conditions. The results are valuable for producing information on the potential integrated management of two-spotted spider mite (TSSM), T. urticae by using acaricides and bio-agents on cucumber plants and their effect on the associated predaceous mite.