India is the world’s largest cotton producer and the only country that grows all four cultivated cotton species. There have been very few studies on the diversity and abundance of natural enemies of cotton insect pests in these cultivated cotton species. Therefore, the current study (2016–2018) was conducted to assess the diversity and abundance of natural enemies that cultivated cotton species harbour. Phule Dhanwantari, Suraj, Suvin, RCH-2, and DCH-32 were the five genotypes used in the study, each with a distinct genetic background. Using the adiv 2.0.1 and vegan R packages, we identified significant differences in natural enemies in terms of species diversity, richness, evenness, and abundance. Analysis of Similarity (ANOSIM) and Non-metric Multidimensional Scaling (NMDS) indicated substantial differences in the natural enemy community structure among the examined genotypes. A total of 17,279 natural enemies were collected and identified across genotypes from seven predatory families and five parasitoid families. The percentage share of these natural enemy families across genotypes and years, in descending order, is Coccinellidae (28.23%) < Tachinidae (19.23%) < Braconidae (12.68%) < Chrysopidae (11.65%) < Chalcididae (9.41%) < Aphelinidae (6.33%) < Pentatomidae (3.29%) < Ichneumonidae (2.37%) < Syrphidae (2.33%) < Vespidae (1.81%) < Asilidae (1.79%) < Geocoridae (0.89%). Coccinellidae, Tachinidae, Braconidae, Chrysopidae, Chalcididae, and Aphelinidae are the six major families that account for more than 85% of all recorded natural enemies. These six families have a higher percentage share in Phule Dhanwantary (90%) compared to the other genotypes. The conservation and better utilization of these natural enemies are crucial for the ecological and safe management of insect pests in the cotton ecosystem.