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. 2020 Sep 17;14(9):e0008591.
doi: 10.1371/journal.pntd.0008591. eCollection 2020 Sep.

Long-Term Mosquito culture with SkitoSnack, an artificial blood meal replacement

Yashoda Kandel  1 , Soumi Mitra  1 , Xavier Jimenez  1 , Stacy D Rodriguez  1 , Alvaro Romero  2 , Brittny N Blakely  2 , Sang-Yeon Cho  3 , Charles Pelzman  3 , Immo A Hansen  1   4
Affiliations

Long-Term Mosquito culture with SkitoSnack, an artificial blood meal replacement

Yashoda Kandel et al. PLoS Negl Trop Dis. .

Abstract

The reliance on blood is a limiting factor for mass rearing of mosquitoes for Sterile-Insect-Technique (SIT) and other mosquito-based control strategies. To solve this problem, we have developed SkitoSnack, a formulated diet for Aedes aegypti (L) mosquitoes, as an alternative for vertebrate blood. Here we addressed the question if long-term yellow fever mosquito culture with SkitoSnack resulted in changed life history traits and fitness of the offspring compared to blood-raised mosquitoes. We also explored if SkitoSnack is suitable to raise Asian tiger mosquitos, Aedes albopictus (L.), and the human bed bug, Cimex lectularius (L). We measured life history traits for 30th generation SkitoSnack-raised Ae. aegypti and 11th generation SkitoSnack-raised Ae. albopictus, and compared them with control mosquitoes raised on blood only. We compared meal preference, flight performance, and reproductive fitness in Ae. aegypti raised on SkitoSnack or blood. We also offered SkitoSnack to bed bug nymphs. We found that long-term culture with SkitoSnack resulted in mosquitoes with similar life history traits compared to bovine blood-raised mosquitoes in both species we studied. Also, Ae. aegypti mosquitoes raised on SkitoSnack had similar flight performance compared to blood raised mosquitoes, were still strongly attracted by human smell and had equal mating success. Minimal feeding occurred in bed bugs. Our results suggest that long-term culture with the blood-meal replacement SkitoSnack results in healthy, fit mosquitoes. Therefore, artificial diets like SkitoSnack can be considered as a viable alternative for vertebrate blood in laboratory mosquito culture as well as for mosquito mass production for Sterile-Insect-Technique mosquito control interventions. SkitoSnack was not suitable to induce engorgement of bed bugs.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Reproductive Performance of SkitoSnack-raised Aedes aegypti.
—(A) Percent engorgement on SkitoSnack (SS) and bovine blood (BB) in different strains. (B) Average number of eggs laid per individual female comparison between SkitoSnack and bovine blood in different strains. (C) Egg hatch rate comparison between SkitoSnack and bovine blood-fed females of different strains. (D) The average weight comparison between 30th generation bovine blood and SkitoSnack-raised mosquitoes. (E) The average wing length comparison between 24th generation bovine blood and SkitoSnack raised mosquitoes. (F) Ae. aegypti engorged on SkitoSnack. The blue color is due to the added food color in the meal.
Fig 2
Fig 2. Female reproductive performance after feeding SkitoSnack or bovine blood to Aedes albopictus.
–(A) Percent engorgement on SkitoSnack (SS) and bovine blood (BB) in different strains. A Kruskal -Wallis H test showed that there was not a statistically significant difference in engorgement rate between treatment groups, (H = 8.69, p = 0.1221) (B) Average number of eggs laid per individual female comparison between SkitoSnack and bovine blood in different strains. (C) Egg hatch rate comparison between SkitoSnack and bovine blood in different strains. (D) The average weight comparison between 11th generation bovine blood and SkitoSnack-raised mosquitoes. Different letters indicate significant difference at P < 0.05. (E) The average wing length comparison between 11th generation bovine blood and SkitoSnack raised mosquitoes. Different letters indicate significant difference at P < 0.05. (F) Ae. albopictus engorged on a SkitoSnack, the green color is due to the added food color in the meal.
Fig 3
Fig 3. Mosquito meal preference.
–(A, upper panel) Experimental setup. The left feeder contains bovine blood, the right SkitoSnack with added green food color. (A, lower panel) The same setup seen from below. (B) Percent engorgement on SkitoSnack (SS) or bovine blood (BB) of mosquitoes raised on these meals. Ns–indicates no significant difference at P < 0.05. (C) Schematic diagram of the experimental setup for Y-tube olfactometer bioassay [21]. The arrow indicates the direction of the air flow. (D) Percent attraction to a human hand in a Y-tube attraction assay of SkitoSnack (SS) raised or bovine blood (BB) raised females.
Fig 4
Fig 4. Flight performance of mosquitoes on the flight mill.
(A) Average velocity of Aedes aegypti males raised on SkitoSnack (SS) or bovine blood (BB). (B) Maximum speed of Aedes aegypti males raised on SkitoSnack (SS) or bovine blood (BB). (C) Average distance traveled of Aedes aegypti males raised on SkitoSnack (SS) or bovine blood (BB). (D) Schematic diagram of mosquito flight mill. For details please see supplemental files.
Fig 5
Fig 5. Fitness of SkitoSnack-raised mosquito males compared to bovine blood-raised males.
(A) Survival curve of mosquito males from SkitoSnack culture and bovine blood culture with and without radiation treatment. The grey box marks the first fourteen days after the start of the experiment that were analyzed as an individual dataset. (B) Experimental setup scheme for the mating competition assay. To avoid a labeling bias, males from both diet groups, SkitoSnack (SS), and bovine blood (BB) were labeled with each stable isotope. On the right side, the three different outcomes we observed are shown. Half of the matings were set up as shown in the upper panel, the other half were set up as shown in the lower panel. (C) Paternity test results. Shown is the number of females that mated with a male from a specific diet group and the number of females that did not mate. No double matings were observed.
Fig 6
Fig 6. Bedbug development after feeding SkitoSnack.
Shown are the (A) percent engorgement, (B) percent of bug nymphs that molted into adults, and (C) percent mortality, seven days after feeding SkitoSnack, bovine blood (Blood), or a PBS solution with 3 mM ATP.
See this image and copyright information in PMC

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