Gentamicin dosing of Dubia Roaches

Today I designed an experiment to observe the effects of gentamicin, a non-cell-permeable antibiotic, on the survival of Blaptica dubia roaches. I will be using this antibiotic in the in vivo growth curve of the LVS strain of Francisella to observe intracellular and extracellular growth. This assay is inspired by a similar experiment that uses Drosophila melanogaster flies found here.

This experiment will include two different methods of antibiotic administration--oral feeding and injection. Future experiments will include the administration of different antibiotics using these methods, so this experiment will offer as a preliminary trial for future work.

Procedure:

1. Solutions of gentamycin were made in PBS and a 50% sucrose solution with a concentration of 1mg/mL.
2. A group of 5 roaches (each 0.7-0.9g) were fed 10µl of the gentamicin in 50% sucrose.
3. A group of 5 roaches were injected with 10µl of the gentamicin in PBS on the 2^nd to last segment of the abdomen on the left dorsal side. Site of injection was cleaned with 70% alcohol before. 
4. Each roach was stored in individual small plastic petri plates in the dark 37degC incubator.
5. Roaches will be administered antibiotic with the same dosing method twice a week for two weeks. Survival will be checked daily. 

//BEE