Table 1.
Statistical analysis summary.
Fig 1.
Prevalence of As. taiwanensis in females over life stages and vertical transmission.
(a) The prevalence of As. taiwanensis oocysts was measured in females. The image was acquired using light microscopy at ×1000 magnification; the black arrow indicates a single oocyst. (b) Prevalence of oocysts in female Malpighian tubules at different physiological stages: unmated (UNMAT), mated (MATED), one day after blood meal (1DABM), and three days after blood meal (3DABM). (c) The prevalence of As. taiwanensis was estimated in larvae in its trophozoïte. The image was acquired using light microscopy at ×400 magnification; the black arrow indicates a single trophozoite. (d) Vertical transmission assessed in newly hatched larvae exposed either to water from oviposition cups (WATER) or to sterile water after egg surface sterilization (EGG SMEARING). Compact letter displays indicate statistically significant differences based on post hoc Tukey HSD tests.
Fig 2.
Impact of As. taiwanensis on female blood feeding, oogenesis, egg laying, egg hatching and larval size.
(a) Abdomen width was measured in parasitized and unparasitized females, before (Unfed) and after a blood meal (Blood fed). (b) Representative image of primary follicle chamber and yolk area in a parasitized female one day after blood feeding (1 DABM), visualized using light microscopy (×100 magnification). (c) Area of primary follicle chambers and (d) yolks was quantified daily from 0 to 3 DABM in parasitized and unparasitized females. (e) Egg-laying dynamics were monitored daily from 3 to 6 DABM in parasitized and unparasitized females and represented as cumulative egg numbers. (f) Hatching success was measured with (2 weeks) and without (0 weeks) a desiccation period in both parasitized and unparasitized groups. (g) Larval size was assessed after hatching for parasitized and unparasitized individuals. Asterisks indicate significant differences (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; Tukey HSD post hoc test). Biorender was used to generate the figure (Licence number TW29FC9MJM). Created in BioRender. Girard, M. (2026) https://BioRender.com/lyuz2tc.
Fig 3.
Transcriptomic impact of Ascogregarina taiwanensis on Ae. albopictus females: Principal Component Analysis and differentially expressed genes (DEGs).
(a) Principal Component Analysis (PCA) of transcriptome profiles from 5 parasitized and 5 unparasitized females at four physiological stages: unmated (UNMAT), mated (MATED), one day after blood meal (1DABM), and three days after blood meal (3DABM). Each dot represents the transcriptome of an individual female. (b) A Venn diagram and an UpSet plot summarize the number and overlap of DEGs between parasitized and unparasitized mosquitoes across female conditions. A total of 9,152 DEGs were identified using pairwise comparisons and Wald tests with Benjamini-Hochberg correction. Intersections are shown as columns. Colors indicate genes specific to parasitized mosquitoes (yellow), unparasitized mosquitoes (grey), or shared between both conditions (dark yellow). Female physiological stages are color-coded and labeled as UNMAT, MATED, 1DABM, and 3DABM. Biorender was used to generate the figure (Licence number TW29FC9MJM). Created in BioRender. Girard, M. (2026) https://BioRender.com/lyuz2tc.
Fig 4.
Functional annotation of differentially expressed genes (DEGs) between parasitized and unparasitized Ae. albopictus females across life stages.
Treemaps display enriched functional categories summarizing Gene Ontology (GO) terms associated with DEGs at four female physiological stages: unmated (UNMAT), mated (MATED), one day after blood meal (1DABM), and three days after blood meal (3DABM). The surface area of each keyword reflects the proportion of associated DEGs. For each stage and infection status, the fold enrichment scores of the top six GO terms are shown (log scale). Percentages of genes involved in each functional category are displayed when exceeding 1%. Biorender was used to generate the figure (Licence number TW29FC9MJM). Created in BioRender. Girard, M. (2026) https://BioRender.com/lyuz2tc.
Fig 5.
Principal Component Analysis and differentially expressed genes (DEGs) in the Ascogregarina taiwanensis transcriptome.
(a) Principal Component Analysis (PCA) of de novo transcriptome profiles from As. taiwanensis within 5 parasitized Ae. albopictus females across four physiological stages: unmated (UNMAT), mated (MATED), one day after blood meal (1DABM), and three days after blood meal (3DABM). Each dot represents the parasite transcriptome associated with an individual host. (b) A Venn diagram and UpSet plot summarize the number and overlap of DEGs identified across stages. A total of 438 DEGs were detected using pairwise comparisons and Wald tests with Benjamini-Hochberg correction. Among them, 378 DEGs were specific to a single life stage (red), while 60 were shared across multiple stages (dark red).
Fig 6.
Functional annotation of Ascogregarina taiwanensis transcriptome across mosquito female life stages.
The As. taiwanensis transcriptome associated with Ae. albopictus females was functionally annotated at four physiological stages: unmated (UNMAT), mated (MATED), one day after blood meal (1DABM), and three days after blood meal (3DABM). GO terms were simplified into functional keywords and visualized as treemaps. The surface area of each keyword reflects the proportion of DEGs it aggregates (log scale). For each life stage, the top six enriched GO terms are shown with their fold enrichment scores (logarithmic scale). Percentages of genes involved in each keyword are displayed when exceeding 1%. Biorender was used to generate the figure (Licence number TW29FC9MJM). Created in BioRender. Girard, M. (2026) https://BioRender.com/lyuz2tc.
Fig 7.
Quantification of proteins, uric acid, and uricase activity in parasitized and unparasitized Ae. albopictus females across physiological stages.
(a) Total protein content, (b) uric acid concentration, and (c) uricase activity were measured in parasitized and unparasitized females at four life stages: unmated (UN), mated (MA), one day after blood meal (1DA), and three days after blood meal (3DA). Measurements were performed on 10 biological replicates per condition, each consisting of three pooled females and normalized to dry mass. Asterisks indicate statistically significant differences (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; Tukey HSD post hoc test).