The use of entomopathogenic nematodes (EPNs) as biological controllers in agricultural crops is an alternative to the use of pesticides of chemical synthesis, which have shown to be harmful to the environment and the human being, and also have lost effectiveness due to the phenomenon of resistance. To meet the demand for EPNs in the field, the in vitro production method has been developed, which, unlike the conventional one (in vivo), has a higher yield and is less expensive. This method requires the design of a culture medium with composition similar to the interior of the insect larvae and the ability of the symbiont bacterium (for Heterorhabditis sp., Photorhabdus luminescens) to digest the biomass of the insect, in this case the components of the medium designed. One of the major difficulties of the method is to determine the inoculation time of infective juveniles (IJs) due to the change of metabolism when growing the bacterium in vitro, where the bacterium goes from being able to digest the medium (phase I) to not being suitable for this function (intermediate phase and II). This capacity is mainly given by the production of lytic enzymes, antibiotics and feeding signals. Based on the foregoing, in this work the kinetics of growth, pathogenicity in Galleria mellonella (Lepidoptera: Pyralidae) and change from phase I to intermediate and II of Photorhabdus luminescens akhurstii SL0708 in in vitro production medium of Heterorhabditis indica SL0708 were evaluated; in order to determine when it is appropriate to inoculate the IJs. To achieve the above, a microbial growth curve was performed in the in vitro production medium of H. indica SL0708 for 96h at 28oC and 150 rpm. By means of a destructive sampling every 12 hours, the formation of biomass (CFU/ml), characteristics of the colonies in NBTA medium, luminescence qualitatively and quantitatively (ALU: arbitrary units of luminescence), pH and reducing sugars equivalent to glucose were determined (g/L). Additionally, an assessment of the assimilation of carbohydrates was made using API 20NE and of the pathogenicity in larvae of the last instar of G. mellonella. Each test was performed in triplicate and the curve was made 3 times in time. As a result it was obtained that the kinetics of P. luminescens akhurstii SL0708 in the evaluated medium consists of an exponential phase of 84 hours with two different growth rates and a diauxic period between these, and a stationary phase after 84 hours. According to the reviewed literature, no studies with similar kinetic results have been reported. On the other hand, it was determined that the difference in growth rates coincides with the change from phase I to intermediate and II, and with the consumption of the majority of glucose (77.4%). This highlights the importance of glucose as a substrate that maintains phase I, which has a growth rate lower than that of the intermediate phase and II. Finally, it was observed that the change from phase I to intermediate and II of the bacterium, has no effect on the pathogenicity of G. mellonella larvae and the assimilation of carbohydrates.