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Bee Bread Sampling Protocol for the National Honey Bee Disease Survey

Please read this protocol carefully and watch the training video 

(https://www.youtube.com/watch?v=i8_gxRbEBOQ) prior to sampling (Please note that 

the video describes how to do pollen sampling, but bee bread sampling is a similar 

procedure. Please see notes below on how to distinguish between fresh pollen and bee 

bread). For additional information or questions concerning sampling methods call the 

University of Maryland Honey Bee Lab at 301-405-7947, or email the UMD Survey 

Coordinator Rachel Fahey at nhbs@umd.edu.

List of equipment:

 Nitrile gloves

 Permanent marker

 Sample vial

 10 wooden sampling sticks

 Aluminum foil

Overview: 

Acute and sub-lethal poisoning from pesticides can be a problem for beekeepers. Honey 

bees come into contact with pesticides outside the colony while foraging for nectar and 

pollen, and inside the colony while feeding, contacting beekeeper applied miticides, or 

contaminated comb wax. Honey bees gather pollen and nectar from many floral sources 

in their environment, store it in their colonies as fermented bee bread and feed it to 

developing brood. Therefore, the pesticides found in sampled bee bread is a snapshot of 

what pesticides that colony was exposed to in their outside environment at that time.

For the 2020 National Honey Bee Disease Survey, bee bread samples will be collected 

for pesticides analysis ONLY from the 5 beekeepers participating in the longitudinal 

monitoring study. State Apiary Inspectors will select and sample 5 apiaries (1 apiary 

from each of the 5 beekeepers) twice for pesticides‒once in the spring (before or at the 

start of the honey flow), and again in the fall after honey flow.

Important: Inspectors should sample the same 8 colonies in the fall that were sampled 

previously in the spring. To achieve this, inspectors need to locate the colonies marked 

with the USDA APHIS survey sticker. If any of the original 8 colonies are dead outs, you 

may take a sample from other colonies in the apiary to bring the total back up to 8 

colonies sampled.
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Procedure:

1. Reminder: Put on a new pair of nitrile gloves prior to sampling each set of 8 

colonies as this prevents cross-contamination between apiaries. 

Figure 1: Bee Bread. Bee Bread is a mixture of pollen and nectar or honey. It can be 

differentiated from fresh pollen by the appearance of a glossy, moist or wet sheen. 

Fresh pollen appears very dry and crumbly.

2. After you have taken the honey bee sample from the brood area for the honey bee 

health survey, look at the frames you have pulled to see if there are any cells with 

bee bread in them. If so, take one end of a sampling stick and insert it all the way to 

the bottom of the cell and rotate the stick all the way around the cell scraping the 

bee bread as you go. You may damage adjacent cells as the stick is slightly larger than 

a cell (Figure 2) but this is normal.

Figure 2: Scraping Bee Bread out of a cell
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3. As you lift the stick from the cell, move slowly as the bee bread, although more wet 

than fresh pollen, is crumbly and may fall off the stick (Figure 3). Place the bee bread 

in the plastic container by scraping the stick on the inside mouth of the vial making 

sure that the bee bread falls into the sampling container. Repeat in at least 4 cells per 

each of the 8 colonies to gather the minimum of 3 grams of bee bread. Use a new 

sampling stick per colony.

4. If the frame you removed for sampling does not have bee bread in it, set it aside and 

try to find another frame with bee bread. If you cannot find any bee bread in the 

colony, move to the next colony for sampling and try to get bee bread from it. If you 

cannot gather bee bread from a particular colony, it is necessary for you to take extra 

samples from the remaining colonies to collect the requisite total of 3 grams.

5. The sampling sticks and gloves can be disposed of in the trash and should not be used 

again.

6. Once all colonies have been sampled, close the vial, and label it with the 

corresponding sample ID (ex. MD-01-2020). Wrap it in aluminum foil to prevent 

sun exposure.

Figure 3: Sampling Bee Bread
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Shipping:

Once all of the colonies in the yard have been sampled for pests and disease, place 

the labeled vials of bee bread into a freezer for storage. Once all 10 samples have 

been collected, ship them to:

University of Maryland Honey Bee Lab

4291 Fieldhouse Drive

Plant Sciences Building Rm. 4112

College Park, MD 20742

It is preferred that these samples be shipped on dry ice, but if not possible, ship them

overnight with cool packs.