Fig. 3: Successive feeding accelerates DENV-2 dissemination and shortens the extrinsic incubation period in both WT and wAlbB Wolbachia-transinfected Ae. aegypti.

a Experimental design for infection and dissemination time course studies of single- and double-fed wildtype mosquitoes lacking Wolbachia. Created in BioRender. Brackney, D. (2025) https://BioRender.com/53u2wpv (WT) and wAlbB mosquitoes. b Percentage of infected single- and double-fed WT mosquitoes 5–10 dpi. Numbers indicate infected mosquitoes over total fed mosquitoes. c Percentage of single- and double-fed WT mosquitoes with disseminated infection 5–10 dpi. Numbers indicate mosquitoes with disseminated infection (measured using legs) over infected mosquitoes. WT single-fed vs double-fed day 5 p = 0.0001. WT single-fed vs double-fed day 6 p = 0.0294. d Percentage of infected single- and double-fed wAlbB mosquitoes 5–10 dpi. Numbers indicate infected mosquitoes over total fed mosquitoes. e Percentage of single- and double-fed wAlbB mosquitoes with disseminated infection 5–10 dpi. wAlbB single-fed vs double-fed day 6 p = 0.0378. wAlbB single-fed vs double-fed day 7 p = 0.0494. Numbers indicate mosquitoes with disseminated infection (measured using legs) over infected mosquitoes. Comparisons were made using two-sided Fisher’s exact tests. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p < 0.0001. Blue = single-fed, red = double-fed. Lines indicate mean ± standard error of the mean of the total sample proportions. Data for time course experiments were collected over 2 replicates. Source data for (b–e) are provided as a Source Data file.