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For the 24-hour period before infection, females were exposed to males of the same genotype in groups of 12 females and 12 males per vial (Day 0 vial).
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Groups of eight virgin females and five males of a given strain or genotype were mated 5 days after eclosion for 3 days, being tipped to a fresh vial each day and then discarded.
Males were transferred individually to new vials on day 3. On day 4 a single female was aspirated into a vial containing a single male, and the pairs were observed for 5 min at 22°.
Females of the genotypes loj/ loj, loj/+, and loj/+ were collected as virgins within 1 2 h of the beginning of the 12 h lighted portion of the light-dark cycle, kept in glass vials in groups of 20 or fewer flies, and aspirated singly into new vials on day 3 after collection.
During the fecundity assay, flies were transferred into fresh food vials every day and the number of eggs laid on the previous day was recorded.
The flies were passed to new vials every day and the number of eggs laid was counted and recorded for the first 24 days of the adult life [ 6].
For treatments involving sexually active males, we combined 5 virgin males with 5 virgin females in a vial on 12th day post egg collection (ie 2-day post eclosion).
Thermotolerance assays were conducted over several days; on each individual day, vials were randomly assigned to six treatments (multiple treatments per day) according to a random block design.
For survival assays preformed at 29oC flies were transferred to new vials every other day during the first 30-40 dand, and then every day for the remainder of the life span.
For each food level, we placed five eggs per clutch in vials with ∼0 PSU Lake Michigan water, fed the animals daily, and observed the vials every other day.
That's why we say it's a pipeline in a vial," she said Wednesday.
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