Abstract
Wolbachia, an obligate intracellular bacterium estimated to infect millions of arthropod species worldwide, is currently being utilized in novel control strategies to limit the transmission of Dengue and Zika viruses. A limitation for Wolbachia-based control approaches is the difficulty of transferring Wolbachia to novel hosts and the lack of tools for the genetic transformation of Wolbachia due to the inability to culture Wolbachia outside the insect host cell in an axenic media. Here, we applied extracellular Wolbachia to phenotypic microarrays to measure the metabolic response of Wolbachia in media formulations with different pH levels and supplementation with Casamino acids. Results suggested a pH of 6.5–6.8 and showed that the supplementation of 1 mg/mL casamino acids increased the survival and longevity of Wolbachia in an axenic medium. In addition, phenotypic microarrays are a useful tool to measure the phenotypic response of Wolbachia under different media conditions, as well as determine specific components that may be required for an axenic medium. This study is an initial step toward the development of a potential Wolbachia axenic culture system.
Highlights
Wolbachia pipientis is an obligate intracellular maternally inherited bacterium found in >55% of insects, as well as filarial nematodes and terrestrial crustaceans [1,2]
Due to the maternal transmission of Wolbachia and the reproductive advantage afforded by Wolbachia, it may be possible to use paratransgenesis to introduce a transgene into a population, in turn producing an antipathogenic molecule to block the transmission of vectored pathogens [8,10,11,12]
To determine that only Wolbachia was in the isolate and there were no residual cells, Aa23 cells isolates were stained with trypan blue
Summary
Wolbachia pipientis is an obligate intracellular maternally inherited bacterium found in >55% of insects, as well as filarial nematodes and terrestrial crustaceans [1,2]. Wolbachia alters host reproduction with several phenotypes, including feminization, parthenogenesis, male killing, and cytoplasmic incompatibility (CI) [3]. The discovery of these reproductive alterations, CI, has caused Wolbachia to be a bacterium of interest for vector control. Wolbachia-induced CI is currently used as part of an incompatible insect technique (IIT) [9] strategy, where males harboring incompatible infections are released with the goal of suppressing natural populations by incompatible mating. Due to the maternal transmission of Wolbachia and the reproductive advantage afforded by Wolbachia, it may be possible to use paratransgenesis to introduce a transgene into a population, in turn producing an antipathogenic molecule to block the transmission of vectored pathogens [8,10,11,12]. The genetic transformation of Wolbachia has not been achieved, due to the difficulty of delivering genetic constructs into an intracellular symbiont
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