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Background Malaria is one of the most lethal infectious diseases in tropical and subtropical areas of the world. Paratransgenesis using symbiotic bacteria offers a sustainable and environmentally friendly strategy to combat this disease. In the study reported here, we evaluated the disruption of malaria transmission in the  Anopheles stephensi-Plasmodium berghei  assemblage using the wild-type (WT) and three modified strains of the insect gut bacterium,  Enterobacter cloacae . Methods The assay was carried out using the  E. cloacae dissolvens  WT and three engineered strains (expressing green fluorescent protein-defensin (GFP-D), scorpine-HasA (S-HasA) and HasA only, respectively). Cotton wool soaked in a solution of 5% (wt/vol) fructose + red dye (1/50 ml) laced with one of the bacterial strains (1 × 10 9 cells/ml) was placed overnight in cages containing female  An. stephensi  mosquitoes (age: 3–5 days). Each group of sugar-fed mosquitoes was then starved for 4–6 h, following which time they were allowed to blood-feed on  P. berghei –infected mice for 20 min in the dark at 17–20 °C. The blood-fed mosquitoes were kept at 19 ± 1 °C and 80 ± 5% relative humidity, and parasite infection was measured by midgut dissection and oocyst counting 10 days post-infection (dpi). Results Exposure to both WT and genetically modified  E. cloacae dissolvens  strains significantly ( P  &lt; 0.0001) disrupted  P. berghei  development in the midgut of  An. stephensi , in comparison with the control group. The mean parasite inhibition of  E. cloacae  WT ,  E. cloacae  HasA ,  E. cloacae  S−HasA  and  E. cloacae  GFP−D  was measured as 72, 86, 92.5 and 92.8 respectively. Conclusions The WT and modified strains of  E. cloacae  have the potential to abolish oocyst development by providing a physical barrier or through the excretion of intrinsic effector molecules. These findings reinforce the case for the use of either WT or genetically modified strains of  E. cloacae  bacteria as a powerful tool to combat malaria. Graphical Abstract

Evaluation of anti-malaria potency of wild and genetically modified Enterobacter cloacae expressing effector proteins in Anopheles stephensi