Milking method’ - Novel Technology for Venom Collection from Aculeate Hymenoptera and used for Screening of In- Vitro Antimicrobial Activity Against Pathogens

Wasps were collected live from the field by using a hand net/sweep net and were maintained in the laboratory under caged condition by providing sugar solution (1:1) along with proteinex. Venom was collected from wasps by ‘milking’. Initially wasps were forced to sting on potato blocks with the help of forceps but the wasps failed to sting on potato blocks. In later trials the wasps were made to sting on sterile paper disc, the wasps were not able to sting on paper disc and did not appear to release the venom either. The wasps were then offered freshly prepared pre-sterilized two per cent agar blocks of approximately 1 cm³. The wasps were able to sting the agar block during this ‘milking’ process and the venom containing agar block was stored in the eppendorf tube containing 50 per cent acetonitrile (ACN) with 0.1 per cent trifluoroacetic acid (TFA). Venom from the selected wasps was collected by ‘milking’ at regular intervals.Then, extract was further concentrated and lyophilized and checked for the biological activity.

‘Miking method’ developed for collection of venom from wasps easily on agar block under laboratory condition and its simple method everybody can use it for venom collection and this can be utilized for conducting different bioassay studies against pathogenic micro organisms.