We used 2% malt extract agar medium plates supplemented
with 0.005% chloramphenicol to isolate yeasts.
Five larvae, a pupa, and two adults were used for yeast
isolation from weevil bodies (Table 2). Three larvae
collected from tree no.2 were dipped in 70% ethanol for
1 min to wet the surface, surface-sterilized for 1 min with
15% hydrogen peroxide (H2O2) solution, dipped again for
1 min in 70% ethanol, and rinsed in sterilized water. First,
the mandibles of one of the larvae were cut and placed on a
plate. Afterward, we cut and opened the abdomen of all the
three larvae and attempted to isolate yeasts from their
body fluid using a sterilized needle. The mandibles of two
mature larvae collected from tree no.4 were also cut and
placed on plates without sterilization. A naturally dead
pupa obtained from tree no.5 was cut, its abdomen opened,
and an attempt to isolate yeasts from its body fluid was
made using a sterilized needle. An adult female was captured
in a building in Kagoshima University. Its rostrum