Japanese encephalitis virus (JEV), a mosquito-borne zoonotic pathogen, is one of the major causes of viral encephalitis worldwide. Previous phylogenetic studies based on the envelope protein indicated that there are four genotypes, and surveillance data suggest that genotype I is gradually replacing genotype III as the dominant strain. Here we report an evolutionary analysis based on 98 full-length genome sequences of JEV, including 67 new samples isolated from humans, pigs, mosquitoes, midges. and bats in affected areas. To investigate the relationships between the genotypes and the significance of genotype I in recent epidemics, we estimated evolutionary rates, ages of common ancestors, and population demographics. Our results indicate that the genotypes diverged in the order IV, III, II, and I and that the genetic diversity of genotype III has decreased rapidly while that of genotype I has increased gradually, consistent with its emergence as the dominant genotype.Japanese encephalitis virus (JEV), a member of the genus Flavivirus in the family Flaviviridae, is a major cause of viral encephalitis and is endemic in several regions of Asia and the Pacific (4, 13), causing an estimated 35,000 to 50,000 infections and 10,000 to 15,000 deaths annually (4, 13, 27). Fifty percent of survivors suffer from lingering neurological effects (7,27,30). Japanese encephalitis (JE) was first reported in Japan in 1924, and JE cases were subsequently reported in many other Asian countries (4,6,7,13,22,27,30). JE was first reported in Australia in 1995 (8, 9, 31). Thus, JE has become a major cause of mosquito-transmitted viral encephalitis on two continents (15,16,25).JEV, the pathogen of JE, has a genome comprising a positive-sense, single-stranded RNA molecule of approximately 11 kb that is capped at the 5Ј end and is not polyadenylated at the 3Ј end. It carries a single open reading frame (ORF) encoding a polyprotein that is processed into three structural (C, M, and E) and seven nonstructural (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins, flanked by 5Ј and 3Ј nontranslated regions (NTRs) (13).Until the latter part of the 20th century, studies indicated that the predominant genotype was genotype III. Since then, there have been multiple reports of genotype I displacing genotype III in many regions (12,18,19,20,24,32,34,35), and in many areas genotype I is now recognized as the dominant strain.As part of a national encephalitis surveillance program, we collected samples from a variety of vectors (mosquitoes and midges), host animals (bats and pigs), and patients with cases of encephalitis in areas where the disease is epidemic, and we isolated viruses from a selection of the JEV-positive samples and sequenced their full genomes. We combined these sequences with other, publicly available full-length genome sequences for a final set of 98 genome sequences. With this set we performed the first detailed evolutionary analysis of JEV based on full-length genome sequences and investigated the epidemiology of genotype I relati...

BackgroundThe current Japanese encephalitis (JE) vaccine derived from G3 JE virus (JEV) can induce protective immunity against G1–G4 JEV genotypes. However, protective efficacy against the emerging G5 genotype has not been reported.Methods/Principal FindingsUsing in vitro and in vivo tests, biological phenotype and cross-immunoreactions were compared between G3 JEV and G5 JEV (wild strains). The PRNT90 method was used to detect neutralizing antibodies against different genotypes of JEV in JE vaccine-immunized subjects and JE patients. In JE vaccine-immunized mice, the lethal challenge protection rates against G3 and G5 JEV wild strains were 100% and 50%, respectively. The seroconversion rates (SCRs) of virus antibodies against G3 and G5 JEV among vaccinated healthy subjects were 100% and 35%, respectively. All clinically identified JE patients showed high levels of G3 JEV neutralizing antibodies (≥1:10–1280) with positive serum geometric mean titers (GMTs) of 43.2, while for G5 JEV, neutralizing antibody conversion rates were only 64% with positive serum GMTs of 11.14. Moreover, the positive rate of JEV neutralizing antibodies against G5 JEV in pediatric patients was lower than in adults.Conclusions/SignificanceLow levels of neutralizing/protective antibodies induced by the current JE vaccine, based on the G3 genotype, were observed against the emerging G5 JEV genotype. Our results demonstrate the need for more detailed studies to reevaluate whether or not the apparent emergence of G5 JEV can be attributed to failure of the current vaccine to induce appropriate immune protectivity against this genotype of JEV.

M. thermoacetica-CdS biohybrid, the first artificial photosynthetic microbial system, has gained a wide variety of scientific attention. Proteomic and metabolomic results indicate that a number of electron carriers and enzymes in the Wood-Ljungdahl pathway play very important roles in electron transfer from CdS to cytoplasm and CO 2 fixation. Targeted metabolomics together with proteome data reveal that glycolysis and the TCA cycle are involved in ATP production in the biohybrid system.