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. 2018 Jul 23;8(1):11023.
doi: 10.1038/s41598-018-29415-5.

The Effect of SkitoSnack, an Artificial Blood Meal Replacement, on Aedes aegypti Life History Traits and Gut Microbiota

Kristina K Gonzales  1 , Stacy D Rodriguez  1 , Hae-Na Chung  1 , Margaret Kowalski  1 , Julia Vulcan  1 , Emily L Moore  1 , Yiyi Li  2 , Stephanie M Willette  3 , Yashoda Kandel  1 , Wayne A Van Voorhies  4 , F Omar Holguin  4   3 , Kathryn A Hanley  1   4 , Immo A Hansen  5   6   7
Affiliations

The Effect of SkitoSnack, an Artificial Blood Meal Replacement, on Aedes aegypti Life History Traits and Gut Microbiota

Kristina K Gonzales et al. Sci Rep. .

Abstract

Public health research and vector control frequently require the rearing of large numbers of vector mosquitoes. All target vector mosquito species are anautogenous, meaning that females require vertebrate blood for egg production. Vertebrate blood, however, is costly, with a short shelf life. To overcome these constraints, we have developed SkitoSnack, an artificial blood meal replacement for the mosquito Aedes aegypti, the vector of dengue, Zika and chikungunya virus. SkitoSnack contains bovine serum albumin and hemoglobin as protein source as well as egg yolk and a bicarbonate buffer. SkitoSnack-raised females had comparable life history traits as blood-raised females. Mosquitoes reared from SkitoSnack-fed females had similar levels of infection and dissemination when orally challenged with dengue virus type 2 (DENV-2) and significantly lower infection with DENV-4. When SkitoSnack was used as a vehicle for DENV-2 delivery, blood-raised and SkitoSnack-raised females were equally susceptible. The midgut microbiota differed significantly between mosquitoes fed on SkitoSnack and mosquitoes fed on blood. By rearing 20 generations of Aedes exclusively on SkitoSnack, we have proven that this artificial diet can replace blood in mosquito mass rearing.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Effectiveness of phagostimulants and meal distribution within the mosquito alimentary canal. ta) Average percent engorgement rates of Ae. aegypti offered bovine blood (control), BSA only (in mAPS buffer and no phagostimulant), and BSA meals supplemented with galactose [50 mM], arabinose [50 mM], trehalose [50 mM], fructose [50 mM], sucrose [50 mM], glucose [50 mM], and ATP [3 mM]. Bars are arranged from weakest to strongest phagostimulant and represent average percent engorgement of at least 3 replicates ± SEM. Different letters indicate a statistical significant difference (p = 0.05 or less). (b–d) Alimentary canal showing crop (cr) and midgut (mg) of (b) females fed on sugar, (c) females fed on blood, and (d) females fed on SkitoSnack (blue hue is the result of the addition of food dye). Scale bar represents 1 mm. (e) Diet distribution of a bovine blood or SkitoSnack meal offered to 12th generation blood-raised or SkitoSnack-raised females. To account for body scaling anomalies, the area of the midgut and crop were divided by individual body weight. Bars represent average area of midgut or crop (mm2) per body weight (mg) ± SEM of least 4 replicates.
Figure 2
Figure 2
Life history traits and microbiota of SkitoSnack mosquitoes. (a) Average number of eggs laid from 11th generation bovine blood- and SkitoSnack-raised females. (b) Egg hatch rate comparison between 12th generation bovine blood- and SkitoSnack-raised females. (c) The average weight comparison between 12th generation bovine blood-raised and SkitoSnack-raised females. (d) The average wing length comparison between 12th generation bovine blood-raised and SkitoSnack-raised females. (e) Composition and percentage of bacterial communities in unfed and 24, 48 and 72 hours post meal (hpm) of bovine blood or SkitoSnack-fed female Ae. aegypti. The pie charts represent taxa at the family level. (f) Principle component analysis. Left panel: females that were unfed or offered a bovine blood meal, middle panel: females that were unfed (same group as in left panel) or offered a SkitoSnack meal, and right panel: merge. (g) Metabolic rate of females and males raised on blood or SkitoSnack. **Denotes statistical significance (P = 0.01).
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