Abstract
Leishmaniasis is a vector-borne parasitic disease transmitted by sand flies that affects 1.3 million people across 98 countries, with limited control strategies due to the lack of an available vaccine and the emergence of insecticide resistance. Novel control strategies that are being explored for mosquito-borne diseases, such as Wolbachia bacterial inhibition of pathogens and genetically modified insects (e.g. using CRISPR-Cas9 editing), rely on the ability to consistently inject embryos of the target species. Here we present a novel method to obtain and inject preblastoderm sand fly embryos of the genus Lutzomyia (Lu.)longipalpis, the principle vector of zoonotic visceral leishmaniasis in South America. The procedures required to obtain sufficiently young Lu. longipalpis colony embryos are described alongside a microinjection technique that permits rapid injection and minimal handling of small sand fly embryos post-injection. Using a strain of Wolbachia as a 'marker' for successful injection, our protocol produced early generation Wolbachia transinfected Lu. longipalpis lines, demonstrating its potential as the first step for use in novel applied strategies for sand fly control.
Highlights
Leishmaniases is a group of vector-borne tropical diseases transmitted by phlebotomine sand flies
The protocol developed here has overcome the first hurdle for egg microinjection in which sufficiently young Lu. longipalpis eggs can be harvested for microinjection
Though the techniques described in this study are a step in the right direction, to “predict” that using Wolbachia will lead to inhibition of Leishmania growth in sand flies based on details of its biology seems a bit of a stretch ( I would like to agree with the authors!)
Summary
Leishmaniases is a group of vector-borne tropical diseases transmitted by phlebotomine sand flies. The causative agent is a kinetoplastid protozoan from the genus Leishmania, which can cause a spectrum of diseases, collectively referred to as leishmaniasis. The clinical symptoms exhibited are influenced by the species of the infecting parasites, the genetic background of the host and associated immunity, human migration and extrinsic factors such as reservoir animal hosts, human migration and control strategies[1]. The spectrum of diseases is referred to as leishmaniases (with an “e”). Human migration is related to transmission of disease and not “extrinsic” factors associated with the clinical symptoms. For the statement of clinical symptoms: “The clinical syndrome exhibited is influenced by the species of the infecting parasites, the genetic background of the host and associated immunity.”. On paragraph 3 of the Intro: In endemic areas where dogs are domestic reservoirs of visceral leishmaniasis (not cutaneous!) It seems to me that the paragraph starting with “Research into novel non-insecticide based control strategies...” should be part of the Discussion
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