Fig 7. Kco_Z infection impacts specific tsetse fitness parameters.

(A) Midgut weight, as an indicator of blood meal digestion proficiency, in six day old Gmm^WT/Kco_Z and Gmm^WT males and females (guts were weighed 24 h after the flies had consumed their last blood meal). Each point on the graph represents one individual, and statistical significance was determined via multiple t-tests. (B). Gonotrophic cycle (GC) length of Gmm^WT/Kco_Z and Gmm^WT females. Age-matched, pregnant females from each group (n = 35 per group) were housed in individuals cages and monitored daily to observe frequency of pupal deposition. Statistical significance was determined via log-rank test. (C) Weight of pupae from Gmm^WT/Kco_Z and Gmm^WT females over three GCs. Each point on the graph represents one individual, and statistical significance was determined via multiple t-tests. (D) Mating competitiveness of Gmm^WT/Kco_Z compared to Gmm^WT males. Matings were setup in individual cages (n = 80). Each cage housed a virgin female, to which one virgin Gmm^WT/Kco_Z and Gmm^WT male was added. Males and females were age-matched, and each had fed twice prior to exposure. Statistical significance was determined via Chi-squared test. (E) Sperm abundance in the reproductive tracts of three and 14 day old (fed twice) virgin Gmm^WT/Kco_Z and Gmm^WT males. Sperm quantity is a reflection of sperm-specific dynein intermediate chain (sdic) transcript abundance. Absolute sdic transcript abundance was determined by comparing experimental sample cycle threshold (C_t) values to those derived from an sdic internal standard curve. Each point on the graph represents one individual, and statistical significance was determined via multiple t-tests. (F) Quantitation of Wigglesworthia and Sodalis in Gmm^WT/Kco_Z and Gmm^WT male and female tsetse. Abundance of symbiont specific gene transcripts (Wigglesworthia, thiC; Sodalis, fliC) was used as a proxy to quantify bacterial load. This was performed by comparing thiC and fliC cycle threshold (C_t) values in Gmm^WT/Kco_Z and Gmm^WT flies to those derived from symbiont gene-specific internal standard curves. Wigglesworthia and Sodalis can be polyploid (Rio et al., 2006; Weiss et al., 2006), and as such, we normalized symbiont genome copy number to constitutively expressed tsetse gapdh copy number. Each point on the graph represents one individual, and statistical significance was determined via multiple t-tests.