BackgroundProtected agriculture (PA) is an alternative allowing the control of environmental variables to produce healthy vegetables. Biofertilizers and biofungicides can reduce the chemical load of pesticides. There is abundant literature documenting individual aspects, such as control of environmental variables, irrigation, biological control, and cost assessments. However, there are no reports documenting integral approaches in which variables are considered altogether in a successful case study of mid-tech technology, suitable in middle-income countries like México. We tested if mid-tech greenhouses using biocontrol and biofertilization can increase profits, using tomato as a model system. This work provides considerations about middle-income countries’ agriculture and the need for a multidisciplinary approach to offer cost-effective, sustainable alternatives to producers. ResultsThis technology yielded up to 254 tons/ha·year of tomato, achieving reductions of 44–60% in water consumption, 25% in chemical nitrogen-fertilization, and 28% in the cost unit of production, increasing the profits by ∼45% in relation to Mexican conventional greenhouses management. ConclusionsThis case study has shown that it is possible to significantly increase profits in mid-tech greenhouse tomato production by increasing productivity and crop quality and decreasing the use of water and agrochemicals through greenhouse automatization, crop management, and beneficial bacteria applied to crops.This manuscript includes a video, supplementary to the main contributions of the project. Please visit this URL:https://youtu.be/uRBGgJqfkLE.How to cite: Serrano-Carreón L, Aranda-Ocampo S, Balderas-Ruíz KA, et al. A case study of a profitable mid-tech greenhouse for the sustainable production of tomato, using a biofertilizer and a biofungicide. Electron J Biotechnol 2022;59. https://doi.org/10.1016/j.ejbt.2022.06.003.