Fig 3. Impact of Spiroplasma infection on the reproductive and nutritional fitness of female tsetse flies.

(A) Gonotrophic cycle (GC) length of offspring from Gff^Spi+ and Gff^Spi- females. Age-matched, pregnant females from each group (n = 34 per group) were housed in individuals cages and monitored daily to observe frequency of pupal deposition. Statistical significance was determined via log-rank test. (B) The weight of pupae deposited by Gff^Spi+ and Gff^Spi- females. Each dot represents an individual pupa, bars represent median values of pupae form each GC. Statistical significance was determined via multiple t-tests with correction via the Holm-Šídák method. (C) Relative densities of Wigglesworthia and Sodalis in Gff^Spi+ and Gff^Spi- females. Relative Wigglesworthia recA and Sodalis rplB copy number was quantified using genomic DNA derived from Gff^Spi+ or Gff^Spi- female midguts (including the Wigglesworthia harboring bacteriome organ). Wigglesworthia recA and Sodalis rplB was normalized relative to Gff gapdh copy number in each sample. Each dot represents one biological replicate (three midguts per replicate), and bars indicate median values. Statistical significance was determined via students t-test. (D) Amount of triacylglyceride (TAG) circulating in the hemolymph of Gff^Spi+ and Gff^Spi- females. Three microliters of hemolymph was extracted from two-week-old pregnant females and TAG was quantified colorimetrically via comparison to triolein standard curve (S2 Fig). Each dot represents an individual pupa, bars represent median values of pupae form each GC. Statistical significance was determined via unpaired t-test.