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. 2022 Mar 3;13:841835. doi: 10.3389/fimmu.2022.841835

Figure 1.

Figure 1

Experimental design and sample collection. Canaries were immunized twice (at day 0 and day 21) with a live vaccine containing E. coli BL21 (n = 7) or E. coli O86:B7 (n =7) or with a mock vaccine (PBS) (n = 3). A group of canaries (n = 5) and part of the birds vaccinated with E. coli BL21 (n = 4) or E. coli O86:B7 (n = 4) were infected with P. relictum at day 14. The birds were used as donors to feed C. quinquefasciatus (n = 20-50 mosquitoes per bird). The midgut of mosquitoes was dissected for microbiota (day 38 and day 45) and oocyst (day 38) analysis and salivary glands for sporozoite (day 45) analysis. Bird sera (used for ELISA and immunofluorescence) and fecal (used for bird gut microbiota analysis) samples were collected at different time points as indicated.