I was thinking of removing the excess bacteria by adding gentamycin to the harvested suspension. but again, the worms might take up the antibiotic and that can kill the bacteria present in the gut of the worm. So I thought of paralysing the worms with levamisole and then add gentamycin to the suspension. For how long the worms can be kept in levamisole solution? would it lead to lyse of the worms if kept for long time? Please help.
Usually, we will treat the worm with 1 mM sodium azide to prevent the expulsion of bacteria from their intestine while doing several washes. we wont give any antibiotic treatments. Protocol from my paper- [Approximately ten worms were transferred to a 2-mL micro centrifuge tube, and the volume was brought to 250 μL with M9 buffer. Silicon carbide particles (400 mg, 1.0 mm; Himedia, India) were added to each tube and vortexed at maximum speed for a minute, which disrupted the worms but did not affect the bacterial survival, and the resulting suspension was serially diluted and plated onto agar (Himedia) plates to determine the CFU].
Thanks a lot! I will try it. Can this be done without the NaN3 step? i mean only washing won't help? will that disrupt the worms?
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