The extensive use of synthetic antibiotics and insecticides in controlling microbes and insect spreads led to a build‐up of resistance strains and caused negative impacts on human health through bioaccumulation in food and the environment. The present study assessed the antimicrobial activity of Amphora coffeaeformis (A. coffeaeformis) ethanolic extract against foodborne pathogenic microbes as well as assessed its ability to control Culex pipiens L. (C. pipiens) larvae. The gas chromatography–mass spectrum (GC/MS) and high‐performance liquid chromatography (HPLC) analysis showed that A. coffeaeformis extract was rich with phenolic compounds, organosulfur compounds, carboxylic acids, amino acids, organic quinoline, dipeptide and monosaccharide. The extract of A. coffeaeformis showed antibacterial capability against all tested bacteria, with minimal inhibition concentration (MIC) values ranging from 20.2 to 66.7 g/mL, as well as antifungal effects against all tested mycotoxigenic fungi, with MIC values ranging from 113.62, 68.95 and 49.37 μg/mL mg/mL. Furthermore, the extract showed high larvicidal activity against C. pipiens larvae at 24, 48, and 72 h, respectively. LC50 values decreased gradually with the treatment period. Likewise, the adult emergence and growth index were significantly negatively related to increasing extract concentrations. Moreover, the extract exhibited a noteworthy depletion in the reproductive potential and increased sterility index of C. pipiens females developed of treated larvae. Besides a reduction in egg hatching percent, increase pupal malformations, increase and adult abnormalities. C. pipiens biochemical markers; glutathione‐S‐transferase, carboxylesterase, acetylcholinesterase, α‐esterase and cytochrome P‐450 showed significant alteration after the extract exposure. Overall, these findings promise the application possibility of A. coffeaeformis extract as a biopreservator against foodborne pathogens and as bioinsecticides for mosquito control as well.