Disease Load Monitoring

Protocol for Real Time Disease Load Monitoring 

(Tier 4 Longitudinal Monitoring)

This project has completed the first two years of the study after the first six month phase began in 2013.  Find the pilot 6 month report at the bottom of this page. 


For Eastern Apiculture Association members please see EAS Disease Load Monitoring Instructions


For Sign-up Instructions Please Fill Out Our Real Time Disease Load Monitoring Registration Form

The protocol presented here has been improved based on the lessons learned from the pilot project which included 22 beekeepers from around the country. It was a successful pilot that allowed us to work out any unforeseen issues. We have summarized the data from the pilot into a 6 month final report which will give beekeepers a better picture of the overall results of this project and allow beekeepers to compare their results to other participants in their region. This report is an example of the individualized report that we only send to participating beekeepers.

Tier 4 Final Reports:

2013 Pilot Year Final Report

2014 Final Report


By monitoring disease levels over time, beekeepers will be able to make better decisions about when to treat colonies and if treatments are effective. Participating beekeepers will be asked to collect samples from 8 colonies once a month over the production season. These samples will be sent to the University of Maryland and processed to determine Nosema and Varroa levels. Each sampling involves opening the eight colonies (the same eight colonies are sampled each period) and removing one frame that contains young, developing brood. Adult bees from this frame are then collected following the standardized method in this document and placed into sample bottles containing alcohol. You will collect two ¼ cup scoops of bees from each hive. You will pour these two scoops of bees into the provided alcohol bottle and seal them. You will repeat this procedure for each of the 8 hives. In summary, you should leave the apiary with eight alcohol bottles full of bees and one data sampling sheet. Finally, you will send the 8 samples to the University of Maryland Diagnostic lab for analysis.

The cost of this ongoing project (subsidized 25% by the Bee Informed Partnership in 2015) for the selected participating beekeepers will be $360 for 6 months (April-September) and then $60 a month for each additional month should you want to extend your season’s sampling.  We also need you to be willing to pay for shipping the samples back to our lab, estimated at less than $10 per month.

This sampling protocol is based off of the USDA APHIS National Honey Bee Survey. For additional information on this effort please visit http://www.aphis.usda.gov/plant_health/plant_pest_info/honey_bees/downloads/sampling_protocol.pdf.

Please read this protocol carefully prior to initiating sampling. For additional information, email askbeeinformed@gmail.com or leave us a message on voice mailbox at 443.296.2470 and we will return your message promptly.


You received 6 months (or 6 sampling periods) worth of sampling material. These kits include:

  • 10 Hive tags
  • 1 Measuring cup (1/4 cup)
  • 1 Funnel
  • 6 Shipping boxes
  • 6 Pre-addressed mailing Labels
  • 6 Sampling Data Sheets
  • 48 Bottles with alcohol (125mL)
  • 6 gallon ziplock bags
  • 1 Sharpie pen
Figure 1: Sampling supplies that will be mailed to you

Figure 1: Sampling supplies that will be mailed to you

You will also need:

  • A staple gun or nails (to affix the hive tags)
  • Postage to return the sample kits (about $10/month)
  • A washtub (optional)

STEP 1: Select 8 colonies

1. Select 8 strong, healthy, queen right colonies located in the same apiary to start the sampling survey. It is important that you select apparently healthy colonies to start with so we can monitor disease appearance and progression.

2. Affix the unique colony tag to each hive (see Figure 2), on the lowest brood box (don’t affix the tag on honey supers). Note that you have received 10 tags. Use only 8 initially and save the spare two tags in the event one or two of the 8 colony dies and you need to tag another colony.

NOTE: IF A COLONY DIES DURING THE SAMPLING PERIOD, REMOVE THE SAMPLE TAG AND Use one of the spare sample tags. If you use the last sample tag, request more. Do not reuse sample tags.

Figure 2: Colony ID Tag

Figure 2: Colony ID Tag

3. Fill out the required information (apiary location, …) on the participant information sheet.

Sampling Steps:

You will need to repeat the following steps every month, around the 15th of the month ± 1 week. We will email you a reminder the week before you need to take your samples.

STEP 2: Sampling in the apiary

1. As you normally would, open the selected colony to the brood nest and examine for disease and queen status/condition. Record any disease/queen status, or unusual conditions present on the sampling data sheet.

2. Remove the lid from one of the sample bottles and place the funnel in the 125 mL bottle filled with alcohol (Figure 3). You might want to widen the opening of the funnel by cutting part of the neck to allow bees to come through more easily.

Figure 3: Sample bottle with funnel

Figure 3: Sample bottle with funnel

3. Find a frame containing brood.

4. Carefully examine the frame to ensure the queen is not on this frame. You don’t want to collect her! We highly recommend that you mark your queen to reduce the risk of accidental sampling.

5. Gently scrape two, ¼ cup scoops of adult bees (about 300) from the brood frame (Figure 4) and place them into the funnel (Figure 5). Gently knock the bottle and funnel to get the bees to fall through the funnel and into the alcohol. 2, ¼ cups of bees should fill more than half of the bottle. Alternatively, if you have a wash tub, shake the bees from the frame into the washtub, gently knock the tub so the bees collect in the corner of the tub and scoop the bees from the tub (Figure 6). Then place the bees into the funnel as described above and shake the remaining bees of the tub back on the colony.

Figure 4: Scooping bees off the brood frame

Figure 4: Scooping bees off the brood frame

Figure 5: Moving bees from measuring cup to alcohol

Figure 5: Moving bees from measuring cup to alcohol

Figure 6: Scooping bees from wash tub

Figure 6: Scooping bees from wash tub

6. Close the bottle tightly; shaking it to make sure the bees are fully dampened with alcohol.

7. Label the sample bottle with your name, date and colony number with the Sharpie marker.

Ex. Jane Bee, 10/15/12, #346

Please note that this colony number MUST match the colony number listed on the Tier 4 data collection sheet you have filled out.

8. Repeat steps one through six until eight colonies have been sampled.

9. Complete the rest of the sampling data sheet to document your management practices of the past month.

STEP 3: Sending the samples

1. Double check that all the lids on the bottles are tightly in place and all bottles are labeled.

2. Place the 8 alcohol bottles (containing bees) into a large Ziploc bag to contain any leaks from the alcohol before placing them into the shipping box (Figure 7).

Figure7: Packaging the 8 sample bottles for return shipment to UMD

Figure7: Packaging the 8 sample bottles for return shipment to UMD

3. Ensure data collection sheets are completely filled out and legible and place in the shipping box.

4. Place the mailing label (Meghan McConnell, University of Maryland, 4112 Plant Sciences, College Park, MD) on the shipping box. Write FROM and your return address on the upper left corner of the box.

5. Email us at askbeeinformed@gmail.com within 24 hours of shipment to notify personnel that an alcohol shipment is expected. 

You should receive a report on the disease levels 2 – 3 weeks from the day the UMD receives your sample. We will send reports both electronically to your email and to your mailing address.

Pollen_TrapWe launched the full project beginning April 2014. One hundred beekeepers managing 10 or more hives throughout the country were eligible to register for a 6 month period of 8 samples a month per apiary. The cost of the full project would be $480, but the Bee Informed Partnership is subsidizing the cost by 25% for this year in 2015. Hence, the cost to you will be $360 for 6 months (April-September) and then $60 a month for each additional month should you want to extend your year’s sampling.

If you have questions or want to sign up to participate, please send an email to askbeeinformed@gmail.com.