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

2015 Final Report


By monitoring disease levels over time you, the beekeeper, 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 sampling season. These samples will be sent to the University of Maryland and processed to determine Varroa and Nosema 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 described in this document and placed into sample bottles containing a salt water solution. You will collect two, ¼ cup scoops of bees from each hive. You will pour these two scoops of bees into the provided sample bottle and cap them. You will repeat this procedure for each of the 8 hives. In summary, you should leave the apiary with eight sample bottles full of bees and one data sampling sheet. You will finally send the 8 samples to the University of Maryland Diagnostic Lab for analysis.

Details of the previous year’s sampling is available on the Bee Informed website at https://beeinformed.org/programs/disease-load-monitoring-2/

This sampling protocol is based off of the USDA AHPIS 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 support@beeinformed.org or leave us a message on voice mailbox at 301-405-3799 and we will return your message promptly. 


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

Materials (Figure 1) Quantity Checklist
Hive Tags 10
¼ Cup Measuring Cup 1
Funnel 1
Shipping Boxes 6
Pre-addressed mailing labels (to UMD Bee Lab) 6
Sampling Data Sheets 6
125 mL bottles with salt water 48
Gallon Zip Lock Bags 6


Sampling Materials

Figure 1: Sampling supplies that will be mailed to you

You will also need:

☐ A staple gun (to affix the hive tag) or nails

☐ Postage to return the samples kits (estimated cost: $12/month)

☐ 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 you select apparently healthy colonies to start with, so we can monitor disease appearance and progression.
  1. Affix the unique colony tag to each hive (see Figure 2). Note that you have received 8 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 FOR A NEW COLONY IN THE SAME APIARY. If you use the last sample tag, request more. Please do not reuse sample tags.

Figure 2: Colony ID Tag

Figure 2: Colony ID Tag

  1. Fill out the required apiary location information on the data collection sheet. 

Sampling Steps:

You will need to repeat the following steps every month, around the 15th of the month ± 1 week.  Try to sample around the same time each month.

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 data information sheet.
  1. Remove the lid from one of the sample bottles and place the funnel in the 125 mL bottle filled with the salt water solution (Figure 3).
Funnel Picture

Figure 3: Sample bottle with funnel

  1. Find a frame containing brood.
  1. Carefully examine the frame to ensure the queen is not on this frame. You don’t want to collect her!
  1. 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 solution. 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.
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 solution.
*Please note that this colony number MUST match the colony number listed on the Tier 4 data collection sheet you have filled out.

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

STEP 3: Sending the samples

  1. Double check that all the lids on the bottles are tightly in place and all bottles are labeled.
  1. Place the 8 sample bottles (containing bees) into a large Ziploc bag to contain any leaks from the solution before placing them into the shipping box (Figure 7).
Final Picture

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

  1. Ensure data collection sheets are completely filled out and legible and place in the shipping box.
  1. 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.
  1. Email us at support@beeinformed.org within 24 hours of shipment to notify personnel that a shipment is expected.

You should receive a report on the disease levels within 2 weeks from the day the UMD lab receives your sample. We will email electronic reports to you each month.

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 support@beeinformed.org.