The entomopathogenic nematode (EPN), Steinernema yirgalemense, is an effective biological control agent against a variety of important insect pests in South Africa. To develop a South African EPN product feasibly in South Africa, EPNs need to be mass-produced. This study aimed to record the population dynamics of S. yirgalemense with in vitro liquid production in shake flasks, with a protein source of powdered egg yolk. The Erlenmeyer flask results indicated variation between flasks, albeit still achieving high yields. The reasons for attaining such variability in the recovery, yield, and growth stages are unclear, hence requiring further studies seeking to increase consistency. The results obtained indicate that, when IJ recovery is low, yields are also low due to relatively few reproductive adults being present in solution, which, in turn, produces more offspring, which later converts to the desired infective juvenile used for product formulation development. For commercial viability, a consistent production system is required that produces predictable yields. This study showed comparable high yields achieved with the flasks and in an early-stage bioreactor setup, being a positive development for S. yirgalemense mass production. Prior to the bioreactor scale-up process, protocol of mass production, the population and growth dynamics of the nematodes in the flask environment requires understanding. This is a positive step, leading to the future commercialisation of a local EPN product.