Honeybee Improvement Program

A Cooperative breeding effort
taking on what Brother Adam called
the "inescapable challenge"
breeding a Varroa mite resistant
strain of honeybee

written by
Jack Griffes
HIP Founder and Breeding Coordinator

The Honeybee Improvement Program (HIP), a Cooperative bee breeding effort, was started in early 1993. Our bee breeding goal is honeybee stock which has under 20% annual loss rate (all causes) while being totally untreated for either of the very troublesome parasitic bee mites, known commonly as the Varroa mite and the Tracheal mite. We started with a stock base that had apparent Tracheal mite resistance. We are currently simultaneously selecting for Varroa mite resistance, high honey production, superior overwintering, and disease resistance. We can see definite improvement. We also fully realize that the high level of genetic resistance we want is probably going to require another 10-20 (or more) years of hard work, with God's help along the way. We are always looking for dedicated beekeepers that both believe genetic resistance is the long term solution to our mite troubles AND are willing to work at developing genetically resistant stock. If you are one of the few that will not only accept but thrive on such a difficult challenge then hop on as a HIP Cooperator. We hope you do.

Enclosed you will find the 1995 HIP protocol - this version contains a good deal of useful info so it will be left available on the Web - we have as of our 24 Oct 1997 HIP Cooperator Meeting modified/updated our basic HIP protocol so as to further streamline. The basic principles we follow are designed to minimize loss while maximizing the gain toward Varroa resistant stock which is our mutual goal. As an example, if you are on pallets you will not be able to run the 10 day weight gain test (unless you are a lot more clever than I - which you probably are). But you can certainly apply the principle of picking out the high producers in some other reliable way (as some or all of you have been doing already). The 10 day weight gain test allows a more rapid assessment of colony productivity which is why I am recommending it be used wherever possible.

1995 HIP Protocol


breeding toward
Varroa mite resistance



By way of analogy you as a HIP Cooperator are a major league scout with your eye always out for players with major league potential. From your A level farm team you make a series of performance based promotions. Each promotion decreases the number of colonies under closest observation. This keeps your scouting efforts focused on colonies that have the greatest potential. The major league superstars become the All Star Team of HIP breeder queens.
Months listed are for Southern Michigan (a Northern state known as "the Great Lake State" in the USA). In parentheses I have listed the critical factors you can use to adjust this calendar to your locale.

March

(very late Winter or ultra early Spring - occasional flight occurs - first pollen) Time to promote the best AAA players to the major league! Distinctly mark untreated test colonies that overwintered strong with lots of stores left. If you are so fortunate as to have too many untreated colonies that are BOTH heavy and strong then check the bottom boards and choose only the ones with clean bottom boards (no hive debris). If these colonies are in several yards (and as mud conditions allow) bring them all to a single yard for further testing under more nearly identical conditions. Since these test colonies will be left untreated you don't want them in treated yards or even within several miles of treated yards that would get rapidly reinfested by their proximity. Leave only these few untreated until other tests are completed. TREAT all other colonies with APISTAN® if you haven't already. This promotion from AAA to major league status helps us select toward several important overwintering traits: frugal use of stores; packing out the brood nest area with stores late in the season; large winter cluster size.

April

(brood patterns improving as pollen is brought in more reliably - flight possible much more often) Time to pick the All Star Team! Run frozen brood HYG (hygienic behavior) test, VKf test, and Varroa population check on the disease free test units. Later in this booklet I will explain how to run these tests so we all do it the same way. The highly HYG colonies that also have a decent to high level of mite chewing are your All Star Team of HIP breeders. They have passed a very difficult test - from here on out baby and coddle them so we can use them as long as possible. Mark AND clip them (if you hadn't already) and keep them in a nuc at home which you check weekly for swarm or supersedure cells. Cut cells and take sealed brood away as needed to keep them from swarming. BUT, keep the nuc strong so they feed the larvae you graft from them very well giving your future queens the best start possible. Give them a frame of drone foundation so you can use their drones for AI and/or to help in your DRONE SATURATION AREA. Treat with APISTAN® as needed. Test results shared with all HIP Cooperators so breeding stock trades can be made.

mid-April or May

- set up your DRONE SATURATION AREA (more info later). Pull APISTAN® strips out 45-56 days after inserting them - (or as long as current label and your flow conditions allow) - minimum of two full brood cycles needed for effective treatment.

May on

- (swarm season - best time to rear queens) Time to begin recruiting the new A level farm team! Graft as large a series of daughter queens as possible from each HIP breeder queen. When you intro a queen (or give a cell) to a production colony write the date and breeder number on the hive front with permanent marker.

Split (or shake) your mating nucs off your Drone Source colonies so you don't bring other drones into your mating area. If you make up strong and well-provisioned nucs early they can build up and make stores IF you have a reliable late flow. Make your nucs stronger with each succeeding batch if you expect them to make their own stores. At the same time, your drone source colonies must remain very strong so take just one brood frame each from several colonies to make up one nuc - I fully realize this is a pain and there are more time efficient ways to make up nucs - in this instance it is imperative to not bring in drones from colonies not worthy of being drone source colonies - you must control both sides of the pedigree to make lasting progress - this requires extra but worthwhile effort.

Another option is to make up an initial batch of nucs. Give each nuc a cell or two. Let the queens get mated and lay for a couple weeks. Then use or bank the queens and give the nucs another cell or marked virgin. Repeat. Restart nucs that die out, abscond, or dwindle by splitting off and swapping places with the stronger nucs. If things go well you end up with two or more times the initial nucs by the end of the Summer. BUT, these nucs probably won't make their own stores nor be strong enough to overwinter without help. This method is geared toward complete requeening in late Summer, which insures you head into Winter with young vigorous queens ready to build up rapidly come Spring.

FEED FUMIDIL-B® to your queen bank and nucs. Don't forget that nucs (fewer reserves and fewer foragers) can starve when larger colonies are persevering - even in the Summer. Treat the nucs with APISTAN® as needed. Varroa infested nucs have a lower percentage of mating success. Before Varroa mites we expected 50 - 75% average mating success over the entire season (from very early to very late). With untreated Varroa infested nucs we got 44% maximum and sometimes under 25% mating success in 1994 and it was dry not wet here. The moral - Treat your nucs with APISTAN®!!!

late June or early July

- participate in annual HIP AI project. Each Commercial size HIP Cooperator brings a minimum of 12 nucs and each Sideliner size HIP Cooperator brings a minimum of 6 nucs for the AI project. In 1996 we did primarily the mixed semen technique on the AI queens - several batches with different mixed semen in each batch. This allows the AI queens to get a full dose of semen and thus be able to produce strong full size colonies that are able to be fully tested. The AI work in 1996 & 1997 was done primarily by Garrett Dodds - I (Jack Griffes) am learning AI of honeybee queens and did some myself as well but not having perfected the technique my level of success is still far below satisfactory (but I keep trying to improve as time permits practice). Each HIP Cooperator pays for the AI work done for his/her own nucs. Once the AI queens are laying the nucs are taken back home by their respective owners - some stay North to winter in nucs - others stay North but winter in full size colonies - still others go South for quicker preliminary testing (and if they pass some test use as potential breeders).

mid-July on

- Time to promote the best A level performers to AA status! Remember the queen in the hive must have been there for over eight weeks before you can test her workers - this is why you wrote the date you requeened on the hive front. Weigh the front and back of each hive and record the results. Ten days later do it again. Figure out which hives in each yard had the largest ten day weight gain AND the largest % of weight gain. Mark these high producers in each yard. Have yards set up to reduce drift so results are accurate. Promote the high producers to AA status by moving them to a new team location. Move them to an isolated yard or yards that has/have a history of good overwintering. They are now potential test colonies so leave them untreated and mark them "DON'T TREAT"especially if employees pull your supers. Set this test yard up with groups of four hives. Each hive in a group must face a different direction. (Ex. North, South, East, West) Separate each group by six feet or more.

After you finish assembling your AA team, run HYG and VKf tests. This is your AA to AAA promotion. Aim for 20 or more test colonies on the AAA team - these will be your best producers which are also your highest in HYG and VKf. Since there is no use in risking loss of colonies that have no chance of becoming breeders your AAA team is all you should leave untreated. Remove from the test yard those colonies with low HYG and VKf test results. DO NOT TREAT THE TEST YARD(S) FOR EITHER MITE IN ANY WAY (chemical, herbal, biological, etc.). That also means no extender nor grease patties because they contain vegetable oil which is one treatment method for tracheal mites. Use Terramycin® mixed with dry powdered sugar instead (if you use Terramycin®) at least on the test colonies so we insure we maintain a high level of tracheal mite resistance while we select for Varroa resistance. Think about it - years from now wouldn't it be awful to discover we now have highly Varroa resistant stock which was being killed by tracheal mites? We can guard against that happening by insuring our test colonies are never treated in any way for either mite. Let's insure we don't lose the ground we have already gained.

Harvest time

- Put in APISTAN® as you pull the last super off each colony (only exception is your AAA test yard which is left untreated.) Treatment of non-test colonies needs to start early enough so that ALL the winter bees develop in Varroa free cells. The later you start treatment the less likely it is to save the colony. Optimum time to begin treatment is around 15 July - early August. You are taking your chances if treatment starts later than mid-August.

Fall

- Colonies headed South to make increase should be TREATED two deeps FULL of bees and VERY heavy (top deep FULL of 100% sealed honey). Combine colonies and give them stores if need be to achieve this. We will send down HIP breeder queens with HIP Cooperators willing to sell cells for use in other HIP Cooperator's nucs. (If you wish to produce and sell HIP cells it is important you let me know ASAP so we have enough breeders selected to go South come Fall.) If you requeen your colonies in the late Summer the increase made come Spring is likely to be greater. Insist that your nuc producer uses HIP cells and marks the hive front with the ID# of the cells using permanent marker or a shot of paint color coded to indicate the breeder queen these cells were grafted from. Use the best of these HIP queen headed hives to stock your DRONE SATURATION AREA.

Frozen Brood HYG test
VKf test

Day 1 - Find a comb that has an excellent sealed brood pattern on both sides in at least one region of the comb. Put a thumbtack in the top bar so you only have to pull this one frame during the rest of the test. Use a serrated steak knife to cut an approximately 2" X 2" sample of sealed brood. The trick is getting a sample that size out of a wired frame. In a wired frame pay attention to where the wires are - cut back to the wire then cut along the wire to get as large a sample as possible. With Duragilt you've got it made just cut right through the plastic center and make the sample any size or shape you like. It's easier to orient the sample if the shape dictates only one way to reinsert it. Close the sample in a ziplock bag prenumbered with the hive ID# written on the bag with permanent marker. Put the numbered bag in a honey jar box to keep the samples in order saving you time when reinserting. After cutting the sample put the thumbtack marked frame in the center of the broodnest no matter where it came from. Put the samples in a Zero degree Faranheit freezer for a minimum of 12 hours but no more than 24 hours. (Under 12 hours at this temperature sometimes results in incomplete kill of the brood - over 24 hours skews the test by causing even non-HYG colonies to clean up the sample.) Put the whole open topped honey jar box in the freezer if room allows - otherwise record the order the samples were in the box so you can put them back in the same order saving you time at the yard. If you forget to do this just set each sample bag in the shade of the hive it goes in before you open any up.

Day 2 - Put the freeze killed brood samples back in the same hive and comb they came from. You have marked the sampled frame top bars with a thumbtack so there won't be any hunting for the proper frame.

Day 3 - 24 hours from when you reinserted the samples, pull your thumbtack marked frame. Look for the queen and gently put her back in the hive if you see her. Shake the bees off the frame. You will see the outline of the brood sample where you cut through brood cells. Now you figure what percentage of the freeze killed sample has been uncapped and removed and record the result. Each whole side of the sample equals 50% - thus if a total of one side's worth of pupae are removed you have 50% - if only half of one sides worth are removed then it's 25% and so on. In other words you make a rapid yet accurate estimate. Return the thumbtacked frame to the center of the broodnest. Look at your recorded results and figure out which half of the test colonies have the best HYG ratings - stick a thumbtack in their hive front to mark them.

Now pull another brood sample to begin a second HYG test on just the best half of your 24 hr tested units. As before you will put these brood samples in numbered bags and freeze them overnight (12-24 hours) (don't let them set out in the sun or even in the shade any longer than it takes you to complete the rest of the test work at that yard - get them in the freezer ASAP).

Now put the sticky-board traps under just the half of the test yard colonies with thumbtacks (or some other mark) to identify them as the highest in HYG. Prepare the sticky-board traps by numbering wax paper sheets (cut to trap size) on both sides (with permanent marker) then spraying top side with aerosol non-stick cooking oil (like PAM or any other vegetable oil spray). When making your own sticky-board traps the critical thing is that the 8x8 mesh (8 wires per inch each way) hardware cloth must be at least 3/8" above the sticky paper surface so the mites can fall through and the bees can't "cheat" by damaging mites stuck on the trap - the damage done to the mites must occur prior to the mites falling. You will get a falsely high amount of mite damage if the bees can reach the wriggling stuck mites because they will try to remove them as if debris even if the bees are low in VKf. Realize that the hardware cloth will eventually sag so it is best to make the traps far deeper than the minimum. As much of the brood nest area as possible (preferably all) must be over the trap so most all the mites that fall hit the trap and get stuck on the sticky paper. I have converted some used 3/4" deep bottom boards into sticky-board traps. Follow the principles and use your ingenuity. Have the trap open toward the back (opposite of entrance) if at all possible so you can service the trap without bees getting under the hardware cloth where they could ruin your test results.

Day 4 - 48 hours from when you reinserted the first frozen brood samples, insert the 2nd frozen brood sample and thumbtack that frame (if a different one) then examine the 1st frozen brood sample and record the results (pull the tumbtack off first sample frame).

Day 5 - Check and record 24 hour results of 2nd HYG test.

Day 6 - Check and record 48 hour results of 2nd HYG test. Pull out the Sticky-board traps. Gently fold the debris laden sticky surfaces together until you have a package that will hold the mites in - then paperclip it to hold it - now you can transport the prenumbered trap papers without fearing loss of mites. Go back to your home or well-lit shop. Record total number of mites on each trap. If there are too many mites to count pick one section and count that section then make a estimate of the total number of mites based on your counted section. Using a 20X hand lens, a dissecting microscope or in a pinch simply a reversed camera lens off a 35mm camera examine 50 mites randomly selected off the diagonals of the posterboard used in the sticky-board traps. I do this by numbering a section of graph paper from 1 to 25 then carefully (so as not to damage them) picking 50 mites randomly off the diagonals of the trap paper and placing them in two rows next to the numbers. I use a toothpick as my tool. Then I look at both top and bottom of each mite carefully and put the SEVERELY damaged mites (large dents, cracked backs, pieces of the mite missing) up a couple of rows and the undamaged ones down a couple rows. The severely damaged ones times two is the percent of SEVERELY damaged mites when you examine 50 randomly selected mites. Record the results. Checking for damaged mites is tedious work, but it will do your heart good to see mites the bees have fatally injured. You may even find yourself cheering the "Varroa killer" bees on. We do not generally bother to look at the VKf level on any hives that did not twice test over 85% HYG in 48 hours. Thus we do not look at all the mites on all the trap papers - we only look at the mites on the papers of the most HYG colonies - this saves much time.

HIP also has an optional TWO YEAR UNTREATED THRIVING SURVIVAL test for those that do not wish to run HYG and VKf tests.

Drift Reduction is crucial



Groups of four hives with each hive facing a different direction minimize drift. Set groups 6 feet or more apart. A very wavy row causes less drift than a straight one - if you don't have room to use a drift minimizing layout and must use drift maximizing straight rows in some yards remember end colonies of rows will be picking up the drifters and thus produce more honey so look for the highest producing non-end colonies and forget the drift skewed end ones if you must use straight rows. Reduce drift and you will reduce disease and parasite spread as well as get more accurate performance test results - you win three ways.

Drone Saturation Area



Drone Saturation (also called drone flooding) means greatly increasing the number of select drones flying in a non-isolated mating area in order to increase the odds of virgin queens (ideally) pure mating (or, more practically, mostly mating) with select drones. Increasing the number of select drones involves moving many drone source colonies into the mating area and managing them to produce extra large numbers of select drones by adding extra drone comb, insuring good nutrition, etc.. When the number of undesirable drones flying in the mating area are simultaneously reduced an even higher level of mating control is achieved. Reducing the number of undesirable drones is a matter of requeening as many managed colonies as possible in a 5-6 mile radius from the mating yard with queens that will produce drones carrying the select genes desired in the matings (this also will increase number of select drones). Typically it also involves eliminating feral colonies, in our case nature (via two mites) has largely done that for us and it is presumed that any remaining feral colonies very likely contain desirable genes (which, if the presumption is correct, also increases select drones). Concentrate your efforts on the two-mile radius surrounding your mating yard and make sure EVERY colony in that area is kicking out loads of the drones you want. Then build on that in future years.

Each drone source colony should have the equivalent of two or three full frames of drone cells to help saturate the area with the best possible drones. You can encourage drone comb building by giving empty frames during swarm season. Put one empty frame between two perfectly straight brood combs during swarm season and it will likely be built predominantly drone comb. Drone Foundation may also be purchased from A.I. Root to provide full combs of drone cells (catalog # H9D - 10 wired Deep sheets for $12.35 (in 1995) - ph. 1-800-289-7668).

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Resource People and Materials:

references abbreviated to save space so more could be included. Read and apply. ABJ= American Bee Journal - GBC = Gleanings in Bee Culture ** (Topic)

Various topics - personal correspondence


Dr. Roger Hoopingarner - Michigan State University (now retired)

Susan Cobey - Ohio State University
1997 Classes At Ohio State Univ.
Queen Rearing May 22-23, 1997
Bee Breeding and Instrumental Insemination June 18,19, 20,1997

Honey Bee Insemination Service
Susan Cobey and Timothy Lawrence
e-mail cobey.1@osu.edu
Specialized Insemination Equipment
Personal Training
Custom Instrumental Insemination Service
New World Carniolan Breeder Stock

Dr. Marla Spivak - University of Minnesota, USA
Dr. John Harbo - USDA/ARS Honey-Bee Breeding, Genetics and Physiology Research Laboratory, Baton Rouge, Louisiana, USA
Brother Adam - Buckfast Abbey, England
Steve Taber - USA
Dr. Ralph Büchler - Germany
Alois Wallner - Austria
Terry Klein - St. Charles, MI, USA
Steve Cantu - Zolfo Springs, FL, USA
Gary Veale - Freeport, MI, USA
Kirk Webster - Middlebury, VT, USA
Dean Breaux - Onsted, MI, USA

Garrett Dodds
Royal Gold Inseminations
Custom Inseminations
Royal Gold Farms
305 E. Hale St.
Ridgeway, OH 43345
937-363-3119
email:honeybees@royalgoldfarms.com



Breeding Varroa Resistant Bees


VARROARESISTENT, by Alois Wallner (Austria)
Imkern heute, by Alois Wallner (Austria)
Aug 1991, ABJ, pgs. 508-510, by Brother Adam
Aug 1992, ABJ, pgs. 530-531, by Steve Taber
Jan 1993, ABJ, pgs.53-54, by Dr. J. Kefuss
March 1993, ABJ, pgs.194-195, by Steve Taber
March 1993, ABJ, pgs. 197-200, by Thomas E. Rinderer, Lilia I. de Guzman, Jovan M. Kulincevic, Gary T. Delatte, Lorraine D. Beaman, Steven M. Buco
Bee World 75(2):54-70 (1994) by Ralph Büchler

Queen Rearing and Stock Selection


VARROARESISTENT, by Alois Wallner (Austria)
Imkern heute, by Alois Wallner (Austria)
May 1988, ABJ, pgs. 341-344, by Susan Cobey and Timothy Lawrence
Jan 1993, ABJ, pgs. 55-57, by Kirk Webster
Beekeeping at Buckfast Abbey, by Brother Adam
Practical Queen Production In The North, by Carl A. Jurica
Nov 1991, ABJ, pgs. 685-687, by Steve Taber
May 1991, ABJ, pgs. 294-296, by G.W. Hayes, Jr.
Feb 1993, ABJ, pgs. 113-114, by Steve Taber
Oct 1991, ABJ, pgs. 626-627, by Steve Taber
May 1992, ABJ, pgs. 327-328, by Steve Taber

Canadian Winter Queen Bank System


March 1993, ABJ, pgs. 201-205, by M.H. Wyborn, M.L. Winston, & P.H. Laflamme

Using controlled natural mating


Sept 1992, ABJ, pgs.603-606, by Gerald M. Loper, Gordon D. Waller, Duana Steffens, and Robert M. Roselle
Sept 1992, ABJ, pg.607, by Leonard H. Hines
Aug 1990, ABJ, pgs.537-542, by Richard L. Hellmich II, & Gordon D. Waller
March 1993, ABJ, pgs. 207-211, by Richard L. Hellmich II, Jorge Ibarra, Manuel Mejia, Thomas E. Rinderer, & Nicholas A. Gutierrez
May 1991, ABJ, pgs. 328-332, by John A. Hogg

Fluvalinate [APISTAN®] after-effects


June 1992, ABJ, pg. 398, by T.P. Liu

Natural colony defense against disease and parasites


June 1993, ABJ, pgs. 431-434, by Jost H. Dustmann

Easy Hive Weighing


Sept 1991, GBC, pgs. 488-489, by James Brimhall

Many topics - must reading


Breeding Super Bees, by Steve Taber
Breeding the Honeybee, by Brother Adam
The Hive and the Honey Bee, 1992 edition, edited by Joe M. Graham

A special THANKS to Dr. Erich Kristen (Toledo, OH) for German to English and English to German translating work he has done. His gracious efforts have made valuable information available that otherwise would not have been. His help in this effort is particularly noteworthy as he is not a beekeeper.

Included below is an update on remaining HIP stock - which is in Garrett's possession. HIP as a cooperative bee breeding program lasted about one decade (historically most bee breeding cooperatives do well to last seven years). Remnants of HIP stock may yet exist in some other HIP Cooperators operations but Garrett's is the most likely to have a high degree of purity due to his location and the AI (II) he has done.

Quoting relevant portions of a Friday 24 Aug 2007 email from Garrett Dodds garrett@royalgoldfarms.com:

Hi Jack,

I thought I (would) give you an update on the HIP Stock that you started.

I have kept the stock going, but not focusing on improving it. Work, family, and church all kept me from the bees as much as I wanted to. Nothing has been treated for the last six years or so, and (they) survive pretty good through the winter. Maybe a 20% loss in the last couple of years. With strong build up in the spring. As I said I didn't work on really improving but maintaining. My focus was on the survivability of the stock with no treatments. No signs of diseases was also looked at. Honey production, gentleness were not really considered. I dropped down to around 20 colonies, so survival and genetic diversity was important. It now survives well, builds up good, is a little aggressive but not bad, and produces well.

(personal portions of email were left out - info about HIP stock is what is being quoted)

. . . starting the first of September. I will be working as a biological sciences tech with the USDA honey bee research lab in Baton Rouge, Louisiana. The reason I am telling you all of this is because as soon as they found out that I had the remaining HIP stock they asked me to bring it down so testing can be done on it to see why and who it is able to survive the mites and to see how well the SMR/VSH trait is maintained over many years of breeding with out selection for it, since you received some of the first SMR stock.

The HIP stock is still going to live on.

God Bless,


Garrett Dodds


----- Original Message ----
From: Garrett Dodds
To: Jack Griffes
Sent: Tuesday, April 29, 2008 5:27:13 PM
Subject: HIP Update


Hi Jack,


I hope things are warming up there in the north. It is very different living down here in the south and keeping bees! Queen rearing has been going on for a couple of months now. I wanted to give you an update on the testing on the HIP stock. Several colonies were at the top of an experiment for SMRi, 3 of the top 5 were HIP queens. SMR was thought to be in two forms SMRi (for initial), and SMRd (for delayed); for mite infertility. SMRd is now called VSH and is the main trigger for the low mite levels. There has been some evidence of an initial reduction in mite infertility and the thought is that it is caused by the brood in some way. Infertility drops in test colonies before the workers from the new queen have started to emerge. What does this mean? We don’t know very early research in this direction. Some HIP colonies were also put in a package test this spring for VSH testing. Interested in seeing how they do. There is starting to be very large scale experiments taking place now, with large commercial beekeepers and following colonies (queens) through normal operations. Some may be taking place up north around Montana .

I will keep you updated on the outcome of the experiments.

Garrett

**************************
Garrett Dodds
USDA Honey Bee Research Lab
1157 Ben Hur Rd.
Baton Rouge , LA 70820 -5502

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Tuesday 29 April 2008

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