Apr 17, 2026
AMBROSIA BEETLE SURFACE STERILIZATION
- M. Elizabeth Moore1,
- Thomas Johnsom1,
- Brian Lovett1
- 1Cornell University
- Moore Entomology Developed Methods
Protocol Citation: M. Elizabeth Moore, Thomas Johnsom, Brian Lovett 2026. AMBROSIA BEETLE SURFACE STERILIZATION. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9bk44l3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 31, 2026
Last Modified: April 17, 2026
Protocol Integer ID: 242370
Keywords: ambrosia beetle surface sterilization surface sterilization, sterilized equipment, large numbers of ambrosia beetle, ambrosia beetle, hundreds of beetle, contaminant fungi outbreak, beetle, events of contaminant fungi outbreak, biosafety cabinet, preparation for experiment
Funders Acknowledgements:
Brian Lovett
Grant ID: USDA NIFA-SCRI 2021-51181-35863
Brian Lovett
Grant ID: USDA-ARS 8062-22410-007-000D
Abstract
Surface sterilization of large quantities of ambrosia beetles can become an arduous and time-consuming endeavor. Elizabeth Moore developed this simple and effective protocol to quickly surface sterilize large numbers of ambrosia beetles in preparation for experiments and in the events of contaminant fungi outbreaks, i.e. Trichoderma. Total time required is <30 minutes for 20 up to hundreds of beetles.
For rearing purposes in bolts, this protocol may be carried out on the bench. For rearing in diet tubes or for experiments, use sterilized equipment and solutions in a biosafety cabinet.
Materials
• 0.01% TWEEN
• 75% Ethanol
• 2% Bleach
• diH2O
• 5 beakers (250 mL is ideal)
• Soft forceps
• Snap ball tea strainer
• Funnel
• Small clip
• 50mL falcon tube
• Paper towels or filter paper
• Sterile petri dish
• Sterile filter paper
Prepare before starting:
Stage beetles in container while preparing for wash.
Prepare/add 75% Ethanol, 2% bleach, 0.01% tween in beakers and add diH2O to two beakers.
Protocol:
Use a funnel to add beetles to a falcon tube, tapping as necessary to prevent escapees.
The tea strainer can comfortably hold 50-100 beetles.
Remove funnel from tube and cover the top of the falcon tube with one side of the tea strainer.
Flip the tube, keep the end covered with the strainer, and tap the beetles out into the strainer.
Close the tea strainer and use a small clip to seal any gaps beetles could get out through.
Repeatedly dunk the tea strainer with beetles inside into the beakers for about ten seconds:
0.01% tween
75% ethanol
2% bleach
diH2O
diH2O
Dab the stainer to dry on a paper towel
Dump beetles into sterile petri dish with sterile filter paper to dry.
Be careful not to crush beetles in the strainer while removing them, use soft forceps to remove any beetles remaining in the strainer. Letting the strainer close may crush beetles and beetles may become stuck in water droplets within the strainer.