Significance: Our work provides evidence that people who are infected with dengue virus without developing detectable clinical symptoms or prior to the onset of symptoms are infectious to mosquitoes. At a given level of viremia, symptom-free people were markedly more infectious to mosquitoes than clinically symptomatic patients. Our results fundamentally change the current paradigm for dengue epidemiology and control, based on detection of dengue virus-infected cases with apparent illness.

A better description of the extent and structure of genetic diversity in dengue virus (DENV) in endemic settings is central to its eventual control. To this end we determined the complete coding region sequence of 187 DENV-2 genomes and 68 E genes from viruses sampled from Vietnamese patients between 1995 and 2009. Strikingly, an episode of genotype replacement was observed, with Asian 1 lineage viruses entirely displacing the previously dominant Asian/American lineage viruses. This genotype replacement event also seems to have occurred within DENV-2 in Thailand and Cambodia, suggestive of a major difference in viral fitness. To determine the cause of this major evolutionary event we compared both the infectivity of the Asian 1 and Asian/American genotypes in mosquitoes and their viraemia levels in humans. Although there was little difference in infectivity in mosquitoes, we observed significantly higher plasma viraemia levels in paediatric patients infected with Asian 1 lineage viruses relative to Asian/American viruses, a phenotype that is predicted to result in a higher probability of human-to-mosquito transmission. These results provide a mechanistic basis to a marked change in the genetic structure of DENV-2 and more broadly underscore that an understanding of DENV evolutionary dynamics can inform the development of vaccines and anti-viral drugs. Dengue virus (DENV) is a mosquito transmitted RNA virus. One of the most characteristic patterns of DENV evolution is that viral lineages, including whole genotypes, are born and die-out on a regular basis. However, the precise cause of such lineage turnover is unclear. To address this issue we explored the genome-scale genetic diversity and evolution of dengue serotype 2 virus (DENV-2) in Viet Nam, a country with a high burden of dengue disease. We observed that DENV viruses assigned to the Asian 1 lineage were likely introduced into southern Viet Nam in the late 1990's (perhaps from Thailand) and subsequently displaced the resident Asian/American lineage serotype 2 viruses. A similar scenario seems to have occurred in Thailand and Cambodia, such that there appears to have been a region-wide proliferation of Asian 1 lineage viruses. Investigation of Vietnamese patients experiencing DENV-2 infection revealed that Asian 1 viruses also attain higher virus levels in the blood than viruses of the Asian/American lineage. This difference in virus titre is likely to have a profound impact on viral fitness by increasing the probability of mosquito transmission, and therein providing Asian 1 lineage viruses with a selective advantage.

Knowledge of the origin and reservoir of the coronavirus responsible for the ongoing COVID-19 pandemic is still fragmentary. To date, the closest relatives to SARS-CoV-2 have been detected in Rhinolophus bats sampled in the Yunnan province, China. Here we describe the identification of SARS-CoV-2 related coronaviruses in two Rhinolophus shameli bats sampled in Cambodia in 2010. Metagenomic sequencing identifies nearly identical viruses sharing 92.6% nucleotide identity with SARS-CoV-2. Most genomic regions are closely related to SARS-CoV-2, with the exception of a region of the spike, which is not compatible with human ACE2-mediated entry. The discovery of these viruses in a bat species not found in China indicates that SARS-CoV-2 related viruses have a much wider geographic distribution than previously reported, and suggests that Southeast Asia represents a key area to consider for future surveillance for coronaviruses.

Despite estimates that, each year, as many as 300 million dengue virus (DENV) infections result in either no perceptible symptoms (asymptomatic) or symptoms that are sufficiently mild to go undetected by surveillance systems (inapparent), it has been assumed that these infections contribute little to onward transmission. However, recent blood-feeding experiments with Aedes aegypti mosquitoes showed that people with asymptomatic and pre-symptomatic DENV infections are capable of infecting mosquitoes. To place those findings into context, we used models of within-host viral dynamics and human demographic projections to (1) quantify the net infectiousness of individuals across the spectrum of DENV infection severity and (2) estimate the fraction of transmission attributable to people with different severities of disease. Our results indicate that net infectiousness of people with asymptomatic infections is 80% (median) that of people with apparent or inapparent symptomatic infections (95% credible interval (CI): 0–146%). Due to their numerical prominence in the infectious reservoir, clinically inapparent infections in total could account for 84% (CI: 82–86%) of DENV transmission. Of infections that ultimately result in any level of symptoms, we estimate that 24% (95% CI: 0–79%) of onward transmission results from mosquitoes biting individuals during the pre-symptomatic phase of their infection. Only 1% (95% CI: 0.8–1.1%) of DENV transmission is attributable to people with clinically detected infections after they have developed symptoms. These findings emphasize the need to (1) reorient current practices for outbreak response to adoption of pre-emptive strategies that account for contributions of undetected infections and (2) apply methodologies that account for undetected infections in surveillance programs, when assessing intervention impact, and when modeling mosquito-borne virus transmission. Most dengue virus infections result in either no perceptible symptoms or symptoms that are so mild that they go undetected by surveillance systems. It is unclear how much these infections contribute to the overall transmission and burden of dengue. At an individual level, we show that people with asymptomatic infections are approximately 80% as infectious to mosquitoes as their symptomatic counterparts. At a population level, we show that approximately 88% of infections result from people who display no apparent symptoms at the time of transmission. These results suggest that individuals undetected by surveillance systems may be the primary reservoir of dengue virus transmission and that policy for dengue control and prevention must be revised accordingly.

Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus mainly spread by Culex mosquitoes that currently has a geographic distribution across most of Southeast Asia and the Western Pacific. Infection with JEV can cause Japanese encephalitis (JE), a severe disease with a high mortality rate, which also results in ongoing sequalae in many survivors. The natural reservoir of JEV is ardeid wading birds, such as egrets and herons, but pigs commonly play an important role as an amplifying host during outbreaks in human populations. Other domestic animals and wildlife have been detected as hosts for JEV, but their role in the ecology and epidemiology of JEV is uncertain. Safe and effective JEV vaccines are available, but unfortunately, their use remains low in most endemic countries where they are most needed. Increased surveillance and diagnosis of JE is required as climate change and social disruption are likely to facilitate further geographical expansion of Culex vectors and JE risk areas.