Characterizing entomopathogenic nematode (EPN) biogeography with a goal of augmentation and conservation biological control requires fine-scale taxonomic resolution, because closely related EPN species can exhibit divergent phenotypes for key properties such as habitat adaptation and insect host specificity. Consequently, we employed high throughput genome sequencing (HTS) to identify and compare EPNs and natural enemies of EPNs in 58 citrus orchards in 2 ecoregions in Egypt (El Beheira and Al Qalyubia governorates). We designed improved primers targeting the ITS2 rDNA to discriminate EPN species and used pre-reported primers targeting D2-D3 region for soil microarthropods. Five EPN species (Heterorhabditis bacteriophora, H. indica, H. taysearae, Steinernema glaseri, and S. scapterisci) and one steinernematid not represented in Genbank databases were detected. This is the first report of S. scapterisci and possibly the unknown (perhaps undescribed) species in Egypt. Only heterorhabditid species, dominated by H. indica, were detected in the reclaimed, sandy desert soils of El Beheira governorate. In the fine textured, ancient farming lands of the Nile delta all six species were detected, but at lower frequency and abundance. Microarthropod family richness (P = 0.01) and abundance (P = 0.001) was higher in the reclaimed lands than in the Nile Delta. Soil clay content, pH and elevation explained significant variation in the mite community structure. Population density of H. indica, the only EPN found consistently and at high abundance in El-Beheira, was inversely related to abundance of species in the nematophagous mite family Rhodacaridae.