Jackman, C. | bioRxiv Science

bioRxiv2020

Endemic and epidemic human alphavirus infections in Eastern Panama, An Analysis of Population-Based Cross-Sectional Surveys

BackgroundMadariaga virus (MADV), has recently been associated with severe human disease in Panama, where the closely related Venezuelan equine encephalitis virus (VEEV) also circulates. In June, 2017, a fatal MADV infection was confirmed in a community of Darien province. MethodsWe conducted a cross-sectional outbreak investigation with human and mosquito collections in July 2017, where sera were tested for alphavirus antibodies and viral RNA. Additionally, by applying a catalytic, force-of-infection statistical model to two serosurveys from Darien province in 2012 and 2017, we investigated whether endemic or epidemic alphavirus transmission occurred historically. ResultsIn 2017, MADV and VEEV IgM seroprevalence was 1.6% and 4.4%, respectively; IgG antibody prevalences were MADV: 13.2%; VEEV: 16.8%; Una virus (UNAV): 16.0%; and Mayaro virus (MAYV): 1.1%. Active viral circulation was not detected. Evidence of MADV and UNAV infection was found near households -- raising questions about its vectors and enzootic transmission cycles. Insomnia was associated with MADV and VEEV infection, depression symptoms were associated with MADV, and dizziness with VEEV and UNAV. Force-of-infection analyses suggest endemic alphavirus transmission historically, with recent increased human exposure to MADV and VEEV in some regions. ConclusionsThe lack of additional neurological cases suggest that severe MADV and VEEV infections occur only rarely. Our results indicate that, over the past five decades, alphavirus infections have occurred at low levels in eastern Panama, but that MADV and VEEV infections have recently increased -- potentially during the past decade. Endemic infections and outbreaks of MADV and VEEV appear to differ spatially. Author summaryPrior to 2010, it was believed that the Madariaga virus (MADV) was primarily associated with equine disease. However, an outbreak reported in Panama, in an endemic area where Venezuelan equine encephalitis virus (VEEV) also circulates, suggested a change in its epidemiological profile. We aimed to reconstruct the epidemiological dynamics of MADV and VEEV, as well as additional alphaviruses known to circulate in the region in order to understand MADV emergence. For this, cross-sectional serosurveys were used to demonstrate that the Alphaviruses MADV, VEEV and Una virus have repeatedly infected humans in eastern Panama over the past five decades. Whilst their historical transmission has been low, we confirm that the transmission has recently increased for both MADV and VEEV.

microbiology↗

bioRxiv2019

Syntrophic co-culture amplification of production phenotype for high-throughput screening of microbial strain libraries

Microbes can be engineered to synthesize a wide array of bioproducts, yet production phenotype evaluation remains a frequent bottleneck in the design-build-test cycle where strain development requires iterative rounds of library construction and testing. Here, we present Syntrophic Co-culture Amplification of Production phenotype (SnoCAP). Through a metabolic cross-feeding circuit, the production level of a target molecule is translated into highly distinguishable co-culture growth characteristics, which amplifies differences in production into highly distinguishable growth phenotypes. We demonstrate SnoCAP with the screening of Escherichia coli strains for production of two target molecules: 2-ketoisovalerate, a precursor of the drop-in biofuel isobutanol, and L-tryptophan. The dynamic range of the screening can be tuned by employing an inhibitory analog of the target molecule. Screening based on this framework requires compartmentalization of individual producers with the sensor strain. We explore three formats of implementation with increasing throughput capability: confinement in microtiter plates (102-104 assays/experiment), spatial separation on agar plates (104-105 assays/experiment), and encapsulation in microdroplets (105-107 assays/experiment). Using SnoCAP, we identified an efficient isobutanol production strain from a random mutagenesis library, reaching a final titer that is 5-fold higher than that of the parent strain. The framework can also be extended to screening for secondary metabolite production using a push-pull strategy. We expect that SnoCAP can be readily adapted to the screening of various microbial species, to improve production of a wide range of target molecules.

synthetic biology↗