Fig 1.
Rhodamine B male body marking.
(A) Example of rhodamine B body marking of male Ae. aegypti compared to an (B) unmarked male. (C) Detection of rhodamine B in a marked male by crushing on filter paper compared to a (D) crushed unmarked male. Rhodamine B marked males were fed on a 0.4% (w/v) rhodamine B honey (50%) solution for 4 d.
Fig 2.
Rhodamine B marking of male seminal fluid.
(A) Example of rhodamine B fluorescence through cuticle of the abdomen of a male Ae. aegypti. (B) Rhodamine B labelling of the testes and accessory glands of male Ae. aegypti. (C) Rhodamine B labelled seminal fluid in the spermathecae and bursa of an Ae. aegypti female transferred during mating with a marked male. (D) Enlarged view of rhodamine B labelled seminal fluid in the spermathecae of an Ae. aegypti that have been opened to reveal the presence of sperm. Males in images A and B were labelled by rhodamine B feeding on a 0.2% w/v solution for 4 days and in image C and D were labelled by rhodamine feeding on a 0.4% w/v solution for 4 days.
Fig 3.
Effect of concentration and feeding time on male body and seminal fluid marking, transference to females after mating, and persistence of seminal fluid-marking after sugar-feeding.
(A) Percent (mean±SE) of Ae. aegypti males with visual body marking after rhodamine B feeding at each concentration (w/v in 50% honey solution) when sugar-fed for 1–4 d. (B) Percent (mean±SE) of females positive for rhodamine B in the spermathecae, bursa or both after mating (24 h exposures) with males fed at each concentration (w/v in 50% honey solution) for 1–4 d. (C) Percent (mean±SE) of females positive for rhodamine B in the spermathecae, bursa or both after mating (24 h exposures) with males marked by rhodamine feeding (0.1–0.8% w/v for 4 d) and withheld from sugar-feeding for a period of 1–3 days.
Table 1.
Efficacy of male body and seminal fluid staining as a function of concentration and length of feeding.
Fig 4.
Male survivorship during and after rhodamine B feeding.
Percent survivorship (mean±SE) of (A) wild type and (B) wMel Ae. aegypti males during and after 4 d of feeding on 0.1, 0.2, 0.4, and 0.8% (w/v in 50% honey solution) rhodamine B solutions relative to unmarked controls (fed 50% honey).
Table 2.
Persistence of rhodamine B labelling of male seminal fluid and body tissue.
Fig 5.
Effect of additional fluids on male body marking by rhodamine B.
The proportion (mean ± SE) of male Ae. aegypti with visual body marking before (no additional sugar-feeding) and after the ingestion of an additional sugar meal (50% honey, no rhodamine B) 48 h after cessation of initial rhodamine feeding.
Table 3.
Effect of secondary sugar-feeding on male body and seminal fluid marking.
Fig 6.
Mating competitiveness of rhodamine B marked males.
Results of mating competitiveness assays represented as the percent (mean±SE) of females mated by (A) wild type and (B) wMel marked Ae. aegypti males vs. unmarked controls. Columns represent treatment means and points represent observations from individual replicates during which 80 virgin females (3–5 d old) were introduced to and kept in the presence of 40 virgin marked and 40 unmarked males (4–5 d old) in a semi-field cage for 72 h.
Table 4.
Results of field releases of rhodamine B marked males.