Background: Prior to the first recorded outbreak of Rift Valley fever (RVF) in Uganda, in March 2016, earlier studies done until the 1970’s indicated the presence of the RVF virus (RVFV) in the country, without any recorded outbreaks in either man or animals. While severe outbreaks of RVF occurred in the neighboring countries, none were reported in Uganda despite forecasts that placed some parts of Uganda at similar risk. The Ministry of Agriculture, Animal Industry and Fisheries (MAAIF) undertook studies to determine the RVF sero-prevalence in risk prone areas. Three datasets from cattle sheep and goats were obtained; one from retrospective samples collected in 2010–2011 from the northern region; the second from the western region in 2013 while the third was from a cross-sectional survey done in 2016 in the south-western region. Laboratory analysis involved the use of the Enzyme Linked Immunosorbent Assays (ELISA). Data were subjected to descriptive statistical analyses, including non-parametric chi-square tests for comparisons between districts and species in the regions. Results: During the Yellow Fever outbreak investigation of 2010–2011 in the northern region, a total sero-prevalence of 6.7% was obtained for anti RVFV reacting antibodies (IgG and IgM) among the domestic ruminant population. The 2013 sero-survey in the western region showed a prevalence of 18.6% in cattle and 2.3% in small ruminants. The 2016 sero-survey in the districts of Kabale, Kanungu, Kasese, Kisoro and Rubirizi, in the south-western region, had the respective district RVF sero-prevalence of 16.0, 2.1, 0.8, 15.1and 2.7% among the domestic ruminants combined for this region; bovines exhibited the highest cumulative sero-prevalence of 15.2%, compared to 5.3 and 4.0% respectively for sheep and goats per species for the region. Conclusions: The absence of apparent outbreaks in Uganda, despite neighboring enzootic areas, having minimal restrictions to the exchange of livestock and their products across borders, suggest an unexpected RVF activity in the study areas that needs to be unraveled. Therefore, more in-depth studies are planned to mitigate the risk of an overt RVF outbreak in humans and animals as has occurred in neighboring countries.

Rift Valley fever (RVF) transmission has intensified in southwestern Uganda since 2016. To quantify human and livestock exposure and associated risks, we conducted a cross-sectional serosurvey in Isingiro, Kabale and Rubanda districts between October and November 2023. A total of 766 humans and 2383 livestock were sampled and tested for RVF antibodies using ELISA, with structured questionnaires capturing demographic, behavioral and environmental data. Human seroprevalence was 11.5% (88/766), varying by district (13.8% Isingiro, 11.8% Rubanda, 6.8% Kabale; p = 0.04). Independent predictors from the multivariate model included raw-meat consumption (aOR 6.11; 95% CI 1.16–27.80), cattle ownership (aOR 2.33; 95% CI 1.27–4.36), male sex (aOR 1.64; 95% CI 1.02–2.66) and younger age compared with ≥50 years (31–49 years: aOR 2.02; 95% CI 1.20–3.48; 18–30 years: aOR 2.37; 95% CI 1.04–5.14). Herd-level seroprevalence was 42.5% (204/480), associated with cattle presence (aOR 6.48; 95% CI 4.10–10.40), lack of carcass burial (aOR 15.70; 95% CI 4.23–63.60), on-farm slaughter (aOR 2.14; 95% CI 1.21–3.89) and increased mosquito activity (aOR 1.75; 95% CI 1.13–2.73). Animal-level seroprevalence was 14.6% (347/2383), highest in cattle (33.8%), with cattle having markedly higher odds than goats (aOR 6.73; 95% CI 4.96–9.14). These findings demonstrate substantial transmission and highlight cattle-centered interfaces as primary targets for control to humans.