Techniques for Experimental Infection of Mosquitoes with West Nile Virus

Liu, Yang; Cheng, Gong

doi:10.1007/978-1-4939-3670-0_13

Cited by 3 publications

(2 citation statements)

References 13 publications

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“…Using the mosquito as an in vivo model can be complex depending on the route used to inoculate the virus. Experimental techniques for WNV infection in mosquitoes vary from thoracic microinjection and membrane blood feeding to blood feeding directly on mice [174]. The origin of the blood source is also an important criterion in the study of vector competence of mosquitoes.…”

Section: Insect Modelmentioning

confidence: 99%

Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts

Fiacre

Pagès

Albina

et al. 2020

IJMS

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West Nile virus (WNV), like the dengue virus (DENV) and yellow fever virus (YFV), are major arboviruses belonging to the Flavivirus genus. WNV is emerging or endemic in many countries around the world, affecting humans and other vertebrates. Since 1999, it has been considered to be a major public and veterinary health problem, causing diverse pathologies, ranging from a mild febrile state to severe neurological damage and death. WNV is transmitted in a bird–mosquito–bird cycle, and can occasionally infect humans and horses, both highly susceptible to the virus but considered dead-end hosts. Many studies have investigated the molecular determinants of WNV virulence, mainly with the ultimate objective of guiding vaccine development. Several vaccines are used in horses in different parts of the world, but there are no licensed WNV vaccines for humans, suggesting the need for greater understanding of the molecular determinants of virulence and antigenicity in different hosts. Owing to technical and economic considerations, WNV virulence factors have essentially been studied in rodent models, and the results cannot always be transported to mosquito vectors or to avian hosts. In this review, the known molecular determinants of WNV virulence, according to invertebrate (mosquitoes) or vertebrate hosts (mammalian and avian), are presented and discussed. This overview will highlight the differences and similarities found between WNV hosts and models, to provide a foundation for the prediction and anticipation of WNV re-emergence and its risk of global spread.

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Section: Insect Modelmentioning

confidence: 99%

Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts

Fiacre

Pagès

Albina

et al. 2020

IJMS

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“…Microinjection has been used to circumvent the midgut when introducing pathogens or nucleic acids, thereby inducing a systemic response 5,6,7,8,9,10 . Several microinjection techniques exist, involving equipment ranging from in-house produced apparatuses that require visual measurement of injection volume to microprocessor-controlled injectors that allow for automated volume delivery as low as 2.3 nL 5,9,10,11 .…”

Section: Introductionmentioning

confidence: 99%

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Sanscrainte

Waits

Geden

et al. 2018

JoVE

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Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are delivered using a non-quantitative method. Accurate delivery of known quantities of nucleic acids or other chemicals is critical to measure the efficacy of the compound being tested and to allow reliable comparison between compounds. Here we provide a reproducible, quantitative microinjection protocol that ensures accurate delivery of specific doses of dsRNA, reducing the mortality typically induced by injection injury. These modifications include the addition of Rhodamine B, a graduated injection needle, and an improved recovery method borrowed from Isoe and Collins. This method allows calculation of dose responses and facilitates comparisons between compounds. Versions of this method have been successfully used on three genera of mosquitoes as well as house flies to assess the reduction in fecundity resulting from gene silencing of ribosomal RNA transcripts. This protocol provides strategies to reduce several challenges of small insect microinjection. Together, mechanical delivery of dsRNAs accompanied by visual verification, identification of effective locations for delivery, and inclusion of a post-injection recovery period ensure accurate dosing and low injury mortality. This protocol also describes an oviposition bioassay for uniform determination of effects on fecundity.

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Animal models of alphavirus infection and human disease

Lucas

Morrison

2022

Advances in Virus Research

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Techniques for Experimental Infection of Mosquitoes with West Nile Virus

Cited by 3 publications

References 13 publications

Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts

Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Animal models of alphavirus infection and human disease

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