These data were collected from a series of experiments testing the effect of bacterial amendments to Aedes aegypti larval rearing water on pupation as well as adult male wing length and survival. The effects of three different genera of bacteria were investigated: Asaia sp., Cedecea sp., and Kocuria sp. Mosquitoes were reared in larval water amended with one of the three bacterial strains or no amendment as a control, and then we measured pupation and collected adults that arose from each treatment and measured their wing length and lifespan. The full experiment was repeated four times. The dataset includes five datasheets. The first is a pupation dataset, where for each individual mosquito assayed, we recorded the day of pupation ("day"). The second is a dataset indicating the overall pupation success (i.e., the proportion of individuals that pupated from each treatment). The third contains the wing lengths of individual males from each treatment measured in milimeters. The fourth is a survival dataset, where for each individual mosquito assayed, we recorded the day of death ("day"). The fifth datasheet contains 16S sequences used to classify the bacterial strains used in the study and the results from NCBI Nucleotide BLAST used to obtain a best match species ID.

Mosquito rearing

We reared Ae. aegypti (Liverpool strain) using standard rearing conditions and rabbit blood (Hemostat, USA). The mosquitoes were maintained at 27°C and a relative humidity of 80% on a 14h light: 10h dark cycle. 

Experimental set-up and bacterial treatment of mosquito larvae

The effects of three different genera of bacteria were investigated: Asaia sp., Cedecea sp., and Kocuria sp. Strains of these genera were previously isolated from Ae. aegypti digestive tracts and have been maintained in 15% glycerol stocks since isolation. We Sanger-sequenced the 16S rRNA gene from each strain using primers fd1 and rp1. Sequences were trimmed and aligned using Unipro UGENE (v 52.1) and submitted to Nucleotide BLAST using default parameters and the nr/nt Nucleotide collection database. This revealed that the best match for Kocuria sp. is Kocuria rhizophila (partial sequence length = 1382bp, query cover = 100%, percent identity = 100%, E value = 0.0), for Asaia sp. is Asaia krungthepensis (partial sequence length = 1351bp, query cover = 100%, percent identity = 99.85%, E value = 0.0) and for Cedecea sp. is Cedecea neteri (partial sequence length = 1403bp, query cover = 100%, percent identity = 100%, E value = 0.0). For each of four independent experiments, four groups were created: a control group consisting of larvae with no bacterial amendment, and three groups of larvae where we amended the larval water with live bacteria from one of the aforementioned genera. Each larval rearing tray was established with 200 first-instar larvae, one liter of reverse osmosis (RO) water, and two pieces of autoclaved orange cat food (each 200-250 mg) (9Lives Indoor Complete dry cat food). The treatment groups then each received an additional 10⁸ Colony Forming Units (CFUs) of one bacterial isolate suspended in 1X PBS, and the control group received a volume of 1X PBS equal to the mean volume of suspended bacteria added to the other treatments. Trays were handled on the open benchtop, and the microbiota was allowed to form conventionally, with the exception that bacterial amendments were added to the three experimental trays. Pupae were transferred to cups and placed in large plastic cages with mesh lids. Adults were allowed to feed ad libitum from a 10% sucrose solution in a bottle located in each container until three days post-peak eclosion, at which point males were separated from females based on standard morphological features and transferred to smaller containers for longevity observation.

Pupation rate and success measurement

Starting on the first day of pupation, pupae were counted daily, and these data were used to estimate the pupation rate. On the last day of pupal counting, overall pupation success was estimated as the total number of pupae obtained per tray divided by the initial number of larvae in the tray (n = 200 for all trays). Larval carcasses were not counted during development; however, at the conclusion of pupal counting, fewer than 6 living larvae remained in any given tray. Thus, all unaccounted for individuals from the initial pool of 200 larvae were assumed to have died during development and were recorded as such.

Wing length measurement

Wing length is a measurement used to estimate the body size of mosquitoes.  Using a binocular microscope (Leica S9i) and subsequent analysis in ImageJ v1.53, the right wings of males from each treatment were measured to the nearest 0.01mm.  Wings were measured from the distal end of the alula to the tip of the R3 vein.

Longevity measurement

Three days post-peak eclosion, all mosquitoes from each container were cold anesthetized, and 25 male mosquitoes from each group were placed into pint-size cardboard cups with mesh lids and allowed to feed ad libitum on a 10% sucrose solution. Each day, dead individuals were removed until no live individuals remained in the cups.