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1.
Sci Adv ; 9(10): eadd7437, 2023 03 10.
Article in English | MEDLINE | ID: mdl-36897949

ABSTRACT

Controlling pathogen circulation in wildlife reservoirs is notoriously challenging. In Latin America, vampire bats have been culled for decades in hopes of mitigating lethal rabies infections in humans and livestock. Whether culls reduce or exacerbate rabies transmission remains controversial. Using Bayesian state-space models, we show that a 2-year, spatially extensive bat cull in an area of exceptional rabies incidence in Peru failed to reduce spillover to livestock, despite reducing bat population density. Viral whole genome sequencing and phylogeographic analyses further demonstrated that culling before virus arrival slowed viral spatial spread, but reactive culling accelerated spread, suggesting that culling-induced changes in bat dispersal promoted viral invasions. Our findings question the core assumptions of density-dependent transmission and localized viral maintenance that underlie culling bats as a rabies prevention strategy and provide an epidemiological and evolutionary framework to understand the outcomes of interventions in complex wildlife disease systems.


Subject(s)
Chiroptera , Rabies virus , Rabies , Animals , Humans , Rabies virus/genetics , Rabies/epidemiology , Rabies/prevention & control , Bayes Theorem , Peru/epidemiology , Livestock , Animals, Wild
2.
Mol Ecol Resour ; 19(1): 128-143, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30240114

ABSTRACT

Microbial communities play an important role in organismal and ecosystem health. While high-throughput metabarcoding has revolutionized the study of bacterial communities, generating comparable viral communities has proven elusive, particularly in wildlife samples where the diversity of viruses and limited quantities of viral nucleic acid present distinctive challenges. Metagenomic sequencing is a promising solution for studying viral communities, but the lack of standardized methods currently precludes comparisons across host taxa or localities. Here, we developed an untargeted shotgun metagenomic sequencing protocol to generate comparable viral communities from noninvasively collected faecal and oropharyngeal swabs. Using samples from common vampire bats (Desmodus rotundus), a key species for virus transmission to humans and domestic animals, we tested how different storage media, nucleic acid extraction procedures and enrichment steps affect viral community detection. Based on finding viral contamination in foetal bovine serum, we recommend storing swabs in RNAlater or another nonbiological medium. We recommend extracting nucleic acid directly from swabs rather than from supernatant or pelleted material, which had undetectable levels of viral RNA. Results from a low-input RNA library preparation protocol suggest that ribosomal RNA depletion and light DNase treatment reduce host and bacterial nucleic acid, and improve virus detection. Finally, applying our approach to twelve pooled samples from seven localities in Peru, we showed that detected viral communities saturated at the attained sequencing depth, allowing unbiased comparisons of viral community composition. Future studies using the methods outlined here will elucidate the determinants of viral communities across host species, environments and time.


Subject(s)
Chiroptera/virology , Metagenomics/methods , Sequence Analysis, DNA/methods , Specimen Handling/methods , Virus Diseases/veterinary , Viruses/classification , Viruses/genetics , Animals , Biodiversity , Feces/virology , Oropharynx/virology , Peru , Virus Diseases/virology
3.
PLoS Negl Trop Dis ; 10(10): e0005048, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27706161

ABSTRACT

BACKGROUND: The outbreak of Zika virus (ZIKV) in the Americas has transformed a previously obscure mosquito-transmitted arbovirus of the Flaviviridae family into a major public health concern. Little is currently known about the evolution and biology of ZIKV and the factors that contribute to the associated pathogenesis. Determining genomic sequences of clinical viral isolates and characterization of elements within these are an important prerequisite to advance our understanding of viral replicative processes and virus-host interactions. METHODOLOGY/PRINCIPAL FINDINGS: We obtained a ZIKV isolate from a patient who presented with classical ZIKV-associated symptoms, and used high throughput sequencing and other molecular biology approaches to determine its full genome sequence, including non-coding regions. Genome regions were characterized and compared to the sequences of other isolates where available. Furthermore, we identified a subgenomic flavivirus RNA (sfRNA) in ZIKV-infected cells that has antagonist activity against RIG-I induced type I interferon induction, with a lesser effect on MDA-5 mediated action. CONCLUSIONS/SIGNIFICANCE: The full-length genome sequence including non-coding regions of a South American ZIKV isolate from a patient with classical symptoms will support efforts to develop genetic tools for this virus. Detection of sfRNA that counteracts interferon responses is likely to be important for further understanding of pathogenesis and virus-host interactions.


Subject(s)
Genome, Viral , Interferon Type I/antagonists & inhibitors , RNA, Viral/genetics , Zika Virus Infection/virology , Zika Virus/isolation & purification , A549 Cells , Animals , Brazil/epidemiology , DEAD Box Protein 58/metabolism , Disease Outbreaks , High-Throughput Nucleotide Sequencing , Host-Pathogen Interactions , Humans , Interferon Type I/biosynthesis , Interferon Type I/genetics , Phylogeny , RNA, Viral/isolation & purification , Vero Cells , Virus Replication , Zika Virus/genetics , Zika Virus/pathogenicity , Zika Virus/physiology
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