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To permit gas exchange leading up to departure from Madagascar (7 10 days after collection), the caps placed on each vial were not securely tightened.
The VVM is a strip on each vial which colour to record the extremes of temperature it has been subjected to in transit.
To collect these offspring each cross was allowed to lay eggs sequentially on three laying vials, spending forty-eight hours on each vial.
Label information on each vial included batch number comprised of serial batch number and "UPIN" (a Unique Patient Identification Number), Study Number, contents labeled as "Recombinant Idiotypic Vaccine" and "New Drug" for investigational use, concentration of 1 mg/mL, date of manufacture, name of sponsor, and name of clinical research organization (CRO) overseeing study conduct.
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Subadults were collected on from each vial daily until they were 8 d after eclosion of the first subadults to count all offspring but not include any grand-offspring emerging in the next generation.
Finally, the climbing index for each vial on the first day of trials is normalized to one.
The effectiveness of various regimens was assessed by charting a "climbing index" for each vial of 20 flies on a longitudinal basis throughout the experiment (Methods S1).
On day 13, each vial was observed hourly for 30 seconds.
On Day 14 each vial was individually subjected to 2.5 minutes of CO2, to simulate the amount of gas normally received when IV vials are mixed, and placed into vials containing fresh medium for oviposition to standard culture densities 25-300 minutes).
The next shipment of drugs was actually pre-packed, with 215 tablets in each vial i.e. for seven months use, but we still needed a pharmaceutical company to put the labels on the vials.
A single fly was maintained in a vial and each vial was tipped on every 1 3 days.
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