Fig. 3

The field A. aegypti mosquitoes breeding in the infectious sewage transmit Zika virus (ZIKV) to hosts. a, b ZIKV survivability in the sewage samples. The supernatant from ZIKV-infected Vero cells was incubated at 28 °C with the sewage samples or phosphate-buffered saline (PBS) (Mock), with either 1 × 10⁵ (a) or 20 pfu/ml (b) of initial titer. The viral survivability was measured over a time course by a plaque assay. a n = 2 biologically independent samples for sewages and n = 3 for the mock. b n = 2 biologically independent samples. c, d Continuous supplementation of ZIKV in the sewage. c Schematic representation. Either the sewage (Sample #5) or PBS (Mock) was used. d ZIKV survivability in the infectious sewage. The mixtures were maintained at 28 °C for a time course and subsequently subjected to a plaque assay. n = 3 biologically independent samples. a–d The data were repeated by two independent biological replicates with the similar results. e Acquisition of ZIKV infection by the A. aegypti Brazil Paraiba mosquitoes breeding in the infectious sewage. Both ZIKV prevalence and infectivity in A. aegypti. The initial ZIKV urine concentration was 20 pfu/ml. f–i ZIKV is transmissible between AG6 mice and mosquitoes breeding in the infectious sewage. f Schematic representation. g, h ZIKV viremia in the mosquito-bitten AG6 mice. g The presence of ZIKV genome in whole blood was assessed using quantitative PCR (qPCR) detection. n represents the mouse number. h Detection by a plaque assay. n = 2 detection replicates. i Prevalence of ZIKV infection in the mosquitoes fed on a viremic AG6 mouse. e, i The mosquitoes for ZIKV detection by TaqMan qPCR. The number of infected mosquitoes relative to the total number of mosquitoes is shown at the top of each column. One dot represents one mosquito. The percentages are represented as the ratios of mosquito infection. a–h Data are presented as the mean ± SEM. a–i The PRVABC59 strain was used for the incubation. Source data are provided as a Source Data file