Abstract

Axenic A. aegypti and A. atropalpus first instars rapidly ingested bacteria. Conventional first instars were hatched in open containers containing distilled water and sterilized diet (Non-sterile). Axenic first instars were hatched in fully sterile conditions and fed sterilized diet (Axenic). Other axenic larvae were fed sterilized diet plus the indicated bacterial isolate. For each treatment, DNA was isolated from a pooled sample of 10 larvae that was collected 6 h post-inoculation after repeated washing and surface sterilization per Coon et al. [21]. DNA was also isolated from cultures of each bacterial isolate. DNA samples were then used as templates with universal or genus-specific primers (see Methods and Table S2). The agarose gel shows ethidium bromide stained PCR products. Lane 1, molecular mass markers labeled in base pairs (bp); Lane 2, universal primers plus DNA from conventional larvae; Lane 3, universal primers plus DNA from axenic larvae; Lane 3-4, Chryseobacterium-specific primers plus template from Chryseobacterium (Control) or axenic first instars inoculated with Chryseobacterium (Larva). The same treatments are then shown for Sphingobacterium (Lanes 5-6), Microbacterium (Lanes 7-8), Leucobacter (Lanes 9-10), Paenibacillus (Lanes 11-12), Aquitalea (Lanes 13-14), and Comamonas (Lanes 15-16). This experiment was repeated four times with independently collected samples and each time yielded identical outcomes for both A. aegypti and A. atropalpus. Table S1. Bacterial isolates used in the study. Table S2. Primers designed and used during the study. (PDF 282 kb)

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