RESUMEN
Objective: To evaluate the potential of local mosquitoes to act as vectors for dengue transmission in Japan. Methods: Serotype 2 ThNH28/93 was used to test the dengue susceptibility profiles of Aedes flavopictus miyarai (Ae. f. miyarai), Aedes galloisi (Ae. galloisi) and Aedes albopictus (Ae. albopictus), which were collected in Japan. We used Aedes aegypti from Thailand as a positive control. The mosquitoes were infected with the virus intrathoracically or orally. At 10 or 14 days post infection, the mosquitoes were dissected and total RNA was extracted from their abdomens, thoraxes, heads and legs. Mosquito susceptibility to dengue virus was evaluated using RT-PCR with dengue virus-specific primers. Differences in the infection and mortality rates of the different mosquito species were tested using Fisher's exact probability test. Results: The infection rates for dengue virus administered intrathoracically to Ae. f. miyarai, Ae. galloisi and Aedes aegypti mosquitoes were identical by RT-PCR on Day 10 post infection. All of the body parts we tested were RT-PCR-positive for dengue virus. For the orally admin-istered virus, the infection rates in the different body parts of the Ae. f. miyarai mosquitoes were slightly higher than those of Ae. albopictus mosquitoes, but were similar to the control mosquitoes (P>0.05). The mortality rates for Ae. f. miyarai and Ae. albopictus mosquitoes were similar (P=0.19). Our data indicated that dengue virus was able to replicate and disseminate to secondary infection sites in all of the four mosquito species (Japanese and Thai). Conclusions: Ae. albopictus is a well-known candidate for dengue transmission in Japan. However, our data suggest that Ae. f. miyarai from Ishigaki Island (near Okinawa Island) and Ae. galloisi from Hokkaido (Northern Japan) should also be regarded as potential vectors for dengue transmission in these regions. Further studies on these mosquitoes should be conducted.
RESUMEN
Fertility is a physiological process of great importance underlying the dynamics of mosquito populations. In transgenesis, it is a prerequisite for the production of subsequent generations and a crucial parameter for evaluating efficiency. Yet, ongoing success in mosquito vector transformation is being severely affected by low embryo survivability. In the prospect of overcoming this impediment, we investigated the darkening/hardening process of the chorion, the effects of some parameters required for transgenesis on hatch success, and erratic hatching in Aedes albopictus, a species that has not yet been targeted for transformation. The eggs from this species, when placed in a moistened environment while whitish, become dark and yet still remain soft approximately 2 h 10 min postoviposition. Those reared in a high moisture environment hatched at a high rate compared with their counterparts submitted to a drier environment. Submission of eggs to p-nitrophenyl-p'-guanidino-benzoate, a substance known to delay the darkening/delay process, resulted in a hatch rate lower than that from eggs soaked in distilled water, which suggests a negative impact on viability. Heat-shock treatment did not taint embryo viability. Overall, eggs displayed a tolerance to an hour of heat shock at 39 degrees C but still hatched at a considerable rate after a 1 hr exposure to 42 degrees C. Hatching was erratic, with a high rate of hatching on the initial flooding and lower rates of hatching on subsequent floodings, all of which resulted cumulatively in considerable hatch success. Our results should serve as a useful reference for the production of both transgenic and laboratory strains of floodwater Aedes mosquitoes.