Honey bees are the only insect which can be instrumentally inseminated. The technique is over 50 years old. It is the only reliable way that the mating of queen bees can be controlled for breeding purposes. Normally a queen will mate in flight with 10 to 20 random drones high in the air and up to several miles away from their home colony.
Today honey bee breeders have selected disease resistant lines of bee, with traits such as Varroa Sensitive Hygiene, or VSH. This trait allows the bees to thrive without chemical treatments by using the bees' own behavior to keep Varroa mites from reproducing. This achievment in bee breeding was only possible with the use of instrumental insemination.
In the above photos, Tom Glenn is collecting the semen from a drone bee, then inseminating a virgin queen with the collected semen from many drones. The process is very humane towards the queen, as she is put to sleep with carbon dioxide during the procedure. She wakes up in a few minutes and within days she is getting on with her job of laying her own weight in eggs every day, about 2,000 eggs. About five million sperm to fertilize all these eggs is stored in an organ called a spermatheca. The sperm stay alive for years until they are used, 5 to 10 at a time when each egg is layed.
Grafting queens is the process of transferring day old female bee larva from the comb of a breeder queen, into an artificial queen cell cup. This is the first step in queen rearing, sort of like planting the seed. The young larva is simply lifted out of her natal cell and placed on a small drop of diluted royal jelly in the queen cell cup.
Any female larva which is less than about 48 hours old since hatching from the egg can be turned into a queen if fed the proper food by the nurse worker bees. But the best queens are raised from the youngest larva, less than 24 hours old, and about 1mm long. Up to about 50 of these grafted queen cells can be placed in a queenless cell builder colony to be fed and raised as queens.
Ten days after grafting the cells are removed from the queen cell builder. The queens will begin hatching out over the next couple of days. Before they hatch, they need to be put into separate colonies or cages to prevent them from fighting. Anytime two queens meet, they will fight to the death of one of them.
The earliest depictions of beekeeping come from the tombs of ancient Egypt from around 2400 BC. Bees were kept in cylinders made from reeds and covered in mud, hives such are still being kept by some beekeepers in the area today. The ancient Egyptian beekeepers also were migratory, putting many hives on rafts floating up and down the Nile River to where the best bee forage was located. Honey was found in the tombs of the pharaohs including King Tutankhamen. The honey recovered from these tombs was still edible, proving that honey never spoils. The pictures above were taken at the Louvre Museum in Paris.
"Science is knowing, art is doing, and common sense is knowing and doing on the basis of experience."Alex Shigo
Over the last few years the beekeeping world has been assaulted by Varroa mites, tracheal mites, and Africanized bees, all of which are problems best solved through bee breeding.The response of the scientific community to these threats has been painstaking research into the natural resistance mechanisms that some bees possess. Today we are confident that several of these resistance mechanisms are effective and practical to breed towards. Bee researchers Marla Spivak and Martha Gilliam sum up the state of the art of breeding for resistance in a recent review of research on hygienic behavior. "Hygienic behavior of honey bees provides multiple benefits for beekeepers with no apparent negative characteristics that accompany the trait......Research has clearly demonstrated the benefits of hygienic bees. Beekeepers should be using this information to improve bee stock." Bee World (1998) The rapid spread of the Varroa mite to beehives worldwide has presented a challenge to bee scientists and bee breeders to develop resistant stock. Both feral and beekeepers' colonies have succumbed by the millions, resulting in loss of pollination of crops, gardens, and wild lands. European honeybees were caught by surprise when the Varroa mite, originally a parasite of the Asian honeybee, jumped to our European honeybee species. The European bees, having no experience with this mite have little defense against the mites. Bee scientists, through careful observation and experiments have discovered several defense mechanisms that are used successfully by the Asian honeybee.
Hygienic behavior - the ability to recognize and remove mite infested larva.
Varroa Sensitive Hygiene (VSH) - a varroa specific hygienic behavior.
Grooming behavior - removing and injuring mites from themselves or another bee.
Short brood development period - resulting in less time for mite reproduction on brood.
Longer time spent on adults - as opposed to inside brood cells reproducing.
Today, miticides are widely used, although Varroa is already showing resistance to these chemicals. Beekeepers long for the good old days when they were not obliged to put miticides in proximity of honey, the most natural of foods. Nearly everyone agrees that the breeding of resistant stock is the best hope for a long term solution to this serious problem.
Given enough time and in the absence of chemical treatment, European bees would probably become adapted to Varroa by natural selection, as the Asian honeybee has. The goal of the bee breeder is to accelerate this process through artificial selection. This is done by identifying the bees with the desired characteristics and controlling their mating to accumulate these traits in a "closed population." Closed population breeding programs have long been used with great success in the breeding of dogs, cattle, and other livestock. It has only been relatively recently that the mating biology, genetics, and techniques in artificial (instrumental) insemination of bees have been worked out so as to make possible, sustainable closed population breeding programs.
Research in the last few years has shown that Varroa resistant traits also exist in European bees. Through selective breeding, hygienic behavior and SMR have now been developed to the point of being in practical use by beekeepers. Today it's encouraging to hear more and more reports of beekeepers able to return to beekeeping without the use of chemicals
The design of a selective breeding program must take into careful account the genetic peculiarities of honeybees.
Honeybees have a haplo / diploid reproductive system. This means males are hatched from unfertilized eggs, so they have no father and only have half the chromosomes of a female.
Unlike most animals, each one of a drone's 10 million sperm are identical clones. Sister bees with the same father share 75% of their genes. This is far more than the 50% found in other species.
The queen naturally mates with up to 20 drones, making the colony a collection of many subfamilies (half sisters with the same mother, but different fathers).
As queen breeders, our role is to take the information gathered by university and government scientists, apply it in our breeding program and then offer our stock to beekeepers. The following are some of the selection tests we are using in our program. See bibliography for more details. 1. Test for Hygienic Behavior There are several methods of testing for hygienic behavior. They all are based on the rate of removal of sealed brood which has been killed behind the capping. The freeze kill method is the most accurate and preferred for scientific work. Liquid nitrogen can be used to freeze kill the brood in a few minutes. Alternatively, a piece of comb can be cut out of the comb and frozen in a freezer. The pin prick method is less accurate but more convenient, it's the easiest method to start checking the hygienic behavior of your bees. Pin prick method Cappings of newly sealed brood cells are punctured with a fine pin to kill the larva beneath. After 24 hours, the number of cells uncapped and cleaned out are counted and recorded. After several replications under different environmental conditions, colonies which have cleaned at least 90% of the cells within 24 hours are considered hygienic. This form of hygienic behavior has been shown to be a significant factor in resistance to Varroa, as well as American foulbrood, and especially Chalkbrood. Genetics of hygienic behavior. 1) Mark a cell directly above three groups of seven newly sealed cells. Use a quick drying paint (e.g. Liquid Paper). Also mark the top bar. 2) Kill all twenty-one larva by pricking them with a pin through the cappings. Use the same hole to prick the larva several times at different angles. 3) Twenty-four hours later count how many cells are completely uncapped and cleaned out. Colonies which have cleaned 19 cells (90%) are considered hygienic.
"The Hygiene Queen" Marla Spivak describes a more scientific method using freeze killed brood.
2. Brood Viability Honeybees can have a lethal gene which can cause spotty brood patterns. The gene which determines the sex of the bee, is called the "sex allele" . Each female has two alleles, one from it's mother and one from it's father. There are 19 or so different variations of the allele possible (let's call them A,B,C...S). The egg and the sperm each contributes one allele. If the combination is different, (e.g. AB, BC, DE, etc.), a female bee results. If two of the same allele variations happen to occur at fertilization (AA, BB, CC..), the egg is eaten immediately upon hatching. This special circumstance creates a male bee known as a diploid drone, which is never allowed to mature in the hive. This is a cause of what beekeepers know as "shot brood," when as little as 50% of the brood survives. Selecting for high brood viability greatly increases the efficiency and productivity of colonies. Low egg viability can be a result of inbreeding and loss of sex alleles. Our closed population breeding program maintains high genetic diversity, thus preventing inbreeding while promoting solid brood patterns.
How to test the brood viability of a queen.
1) Cut a parallelogram from a card. There should be 10 worker cells per side, enclosing a total of 100 cells. 2) Place template over the most solidly sealed patch of brood. 3) Count the number of empty cells. Subtract from 100 to get the percentage of brood viability. 4) In this example 100 - 13 = 87% viability. Above 85% is acceptable.
3. Temperament Our first method is a standardized test given to all colonies in the apiary under the same conditions to be sure gentle temperament is maintained. After an apiary with many hives has been worked, it is often impossible to determine which hives the more aggressive bees came from. A second technique we find useful to separate out any "mean" bees is to capture them in a black plastic bag. Do this by swinging it's open end in a figure eight in front of you, this both attracts and captures the bees. Once captured, the bees can easily be immobilized with carbon dioxide gas or refrigeration and then marked with paint. After being released these guard bees can be found at the entrances of the hives which are then culled from the program because of unacceptable temperament. We select drones and queens from only gentle, workable colonies. 4. Tracheal Mite Resistance (graphic) The mechanism of resistance against this mite is as yet unknown, but it does clearly exist. Recent evidence suggests that grooming behavior as the mites migrate from one bee to another may be a means of control. Fortunately this trait appears to be controlled by dominant gene(s) and occurs widely in honey bees. Thousands of bees are examined to determine which hives show resistance. A test can also be done by placing newly hatched workers from various hives among bees known to be infested, for about a week. (Bees are only susceptible to mite infestation when they are very young, up to ten days old). The bees are then dissected and mites in the trachea are counted. Differences in attractiveness or susceptibility to mite infestation are evident. 5. Honey Production and Comb Building Comparisons of honey production and comb building are made of individual colonies in the same apiary and under the similar conditions. High honey production results from the right number of healthy bees being in the hive at the proper time. Factors enhancing health will likely increase honey production. High honey production using disease resistant stock is the ultimate goal of our breeding program
Instrumentally Inseminated Breeding Stock
Why use instrumentally inseminated queens in a breeding program?
Honeybees are the only insect which can be instrumentally inseminated, allowing bee breeders to have complete control over the mating of the queens and drones. Instrumental insemination allows two very powerful procedures which are not possible with naturally mated queens.
1. Queens can be mated to a single drone, simplifying selection for specific traits.
2. Queens can be mated to hundreds of drones, maintaining genetic diversity.
Breeding for specific traits using single mated queens
The multiple mating habits of bees has always been an obstacle to progress in breeding bees for specific traits.
To simplify the selection process, queens can be instrumentally inseminated to single drones.
All 10 million sperm produced by a single drone are identical clones.
Queens mated to a single drone produce progeny with extreme consistency.
Genetic consistency and genetic diversity are opposite ends of a spectrum. One necessarily gives up diversity in trade for "fixing" any trait in an individual, a colony, or a population.
This genetic trade-off can be optimized using single drone inseminations together with mating other queens with large numbers of drones (supermated).
Conserving genetic diversity using supermated queens
Naturally mated queens normally mate with from ten to twenty drones on their nuptial flights.
Oddly enough, the semen carried in a single drone is more than enough to fill the queen's spermatheca, where sperm are stored for the lifetime of the queen.
The queen takes a great risk to gather so much extra genetic diversity for her progeny, to the advantage of her colony.
Recent findings suggest that bees of different patrilines specialize in the various behaviors found in honeybee colonies.
It is thought that the more genetic options, the more behaviors will be optimized.
Using instrumental insemination enables the breeder to go one step beyond nature and inseminate the queen with hundreds of drones.
Drones are chosen from colonies expressing desirable traits such as disease resistance, high honey production and gentleness.
Semen is extracted from hundreds of drones from many colonies, mixed together, then used to inseminate many queens. We call these queens "supermated".
Supermated queens have very high brood viability due to the high diversity of sex alleles, which means more bees in their colonies.
By maintaining a high degree of genetic diversity the negative effects of inbreeding are avoided.
Supermated queens contain many times the genetic diversity of naturally mated queens. They are excellent as breeder queens in breeding programs to prevent unintentional inbreeding.
To keep things simple, further examples will show a queen mated to a single drone.
This can be done with instrumental insemination, to sort genetic traits.
Number of chromosomes in bees. A key factor.
Drones result from unfertilized eggs (parthenogenesis).They have no father.
All eggs and sperm carry 16 chromosomes each.
Each egg contains a unique combination of 50% of the queens genes.
All 10 million sperm produced by a drone are identical clones.
Since each queen mates with 10-20 drones, colonies are comprised of subfamilies, each having the same mother but different fathers.
Workers of the same subfamily are related by 75% of their genes.
This "extra" close relatedness may explain the cooperative, and altruistic behaviors found in colonies.
It also explains why workers forego their own reproduction in favor of helping their queen mother raise more sisters. Their sisters are more closely related to them than their own offspring would be. (75% vs 50%)
Cordovan color
Cordovan Italian
Wild type Italian
Hygienic behavior
Recent research suggests that this model of hygienic behavior may be too simplistic. Evidence now shows that there may be as many seven genes involved.
Day 1 - Give breeder hive an empty dark brood comb to lay eggs in.
Day 4 - Transfer (graft) larva into artificial queen cell cups, from the breeder comb. Place the frame into a strong colony (cell builder) made queenless the day before.
Day 14 - Remove completed cells from cell builder. Leave one cell behind to replace the queen. Keep queen cells warm (80-94 F) until they are placed in queenless hives (mating nucs).
Day 22 - Virgin queens are ready to mate. They require nice weather (69 F), and an abundance of drones to mate with. A few colonies within a mile are adequate for providing drones for mating.
Day 27 - If queens mate without weather delay, they should now be laying eggs.
Weather delays in mating will add days to the process, after 3 weeks delay, virgin queens may start to lay unfertilized eggs.
Time your activities so that warm temperatures and drones are available when the queens are ready to mate.
How to graft queens
Grafting is simply the process of transferring larva from the worker cell of the breeder's hive to an artificial queen cell. The shape of the cell, along with the queenless condition of the hive receiving the newly grafted cells stimulates the workers to feed them a diet which make them develop into queens. A grafting tool can be as simple as a bent piece of wire, or several varieties can be purchased. The tool is slipped under the larva which is lifted out and placed in the bottom of the queen cell cup. Priming the cells with a small drop of royal jelly or even diluted honey makes it easier to float the larva off the tool. Don't flip over the larva. An unsteady hand is helped by bracing it lightly on the comb. Good light is essential, a headlamp works well, sunlight is ok if done quickly. Magnifying lamps are useful for those with poor eyesight. Some people are expert grafters from the start, others need more practice. Grafting is what prevents most people from attempting queen rearing. This is unfortunate, because with an hour or two of practice, anyone can acquire this valuable skill. Give it a try.
The Breeder hive
Graft from your best colony, or purchase a selectively bred breeder.
Use the youngest (smallest) larva.
By placing an empty brood comb in the brood nest 4 days before you graft, the larva will be the right age.
Setting up the Cell Building Colony
Any strong hive can serve as a cell builder.
Remove the queen one day before you graft cells.
Place grafted cells in center of the brood nest.
Place about 30 cells per colony.
Large cells will be produced by well nourished colonies.
Feeding is not necessary if a light honeyflow is on and pollen is abundant.
Setting up Mating Nucs
Cells are placed in queenless colonies the day before they hatch.
The mating process is usually only 75% successful.
Small mating colonies minimize the losses due to unsuccessful queens.
Mini-nucs are convenient for raising large numbers of queens.
Cells can be placed in any queenless colony.
Most queenless colonies will accept cells without queen cell protectors.
Recent research shows that queen quality is best when they are left to lay eggs in the nuc for about a month.
Mating conditions
Queens are ready to mate 5-7 days after hatching.
Temperature must be at least 69 F with no strong winds.
Virgin queens mate with 10 to 20 drones on one or more flights.
Drones and queens may fly a mile or more to drone congregation areas.
Queens will begin laying eggs 2 to 4 days after mating.
Mating can be delayed up to 3 weeks and still be successful.
USDA scientists at the Honey Bee Breeding, Genetics & Physiology Laboratory in Baton Rouge, Louisiana, have defined and tested a trait of the honeybee that is a Varroa Sensitive Hygienic behavior (VSH). Reproduction of varroa mites triggers their removal by the bees. Formerly known as bees with SMR, Suppressed Mite Reproduction.More...
These golden yellow Italians are excellent for commercial pollination as well as honey production. Our Pol-line hygienic Italian breeders have incorporated Varroa Sensitive Hygienic behavior, VSH for enhanced resistance to varroa mites.
Early Spring build up
Large clusters
Great brood production
Excellent for commercial pollination and honey production
Cordovan color is a genetic trait controlled by a single recessive gene, similar to blue eyes in humans. It is a color variation rather than a race of bee.The cordovan gene changes black body parts (head, thorax, bands) to reddish brown. The abdomen is golden yellow to the tip.Their bright color makes them the easiest queen to find. These are our most gentle bees. We have crossed our cordovan bees with VSH bees to enhance their Varroa mite resistance.More... .
Carniolans are known for their winter hardiness and rapid spring build up.They are excellent honey producers and comb builders. They have proven to be exceptionally resistant to tracheal mites.This is our darkest colored line of bees. Our Carniolans have been crossed with VSH bees for enhanced resistance to Varroa mites.
We also offer pure VSH queens crossed with Italian or Carniolan drones. These are especially useful for programs utilizing instrumentally insemination, since the drones produced by these queens will be pure VSH.
Shipping - Queens are sent via UPS Next Day Air. Add $50.00 shipping per shipment. Queens are shipped in 3 hole candy introduction cages with attendant bees. A push-in cage is included for each queen. JZs BZs plastic cages are available upon request. Please send your street address as UPS does not send to Post Office Boxes.
.All queens are instrumentally inseminated and marked with a numbered disk. Also one half of one forewing is carefully clipped. International queen color code:
.Questions & Answers
Are your queens fertile? Yes. All of our queens are now instrumentally inseminated. A queen is shipped only after we are sure she is fertile and laying a good brood pattern. Why use instrumentally inseminated queens? Naturally mated queens normally mate with 10-20 random drones, sometimes miles away from their hives. Instrumental insemination is necessary to apply the principles of selection and breeding to bees. Controlling the mating allows us to provide you with disease resistant and highly productive queens. When are queens available? Instrumentally inseminated queens are available from March to December. Supplies are limited in the early spring. What about mites and diseases? Our stock has been proven to have tracheal mite resistance and hygienic behavior. We take preventive measures to insure that mites or disease are not present on the bees we sell. Fumagilin-B is included in the queen candy as protection against Nosema. A State of California Health Certificate is included with each shipment.
USDA scientists have imported a strain of honeybee from Russia which are naturally resistant to Varroa mites. The imported queens were selected from bees which had been exposed to mite infestation for nearly a century. If natural selection can produce tolerance to Varroa, then these bees may be our best bet. The new Russian strain has gone through a quarantine and testing period since 1997, and are now being released to American beekeepers. Ongoing selection for further resistance to Varroa is being carried out by a team of top USDA scientists from the Honey Bee Breeding ,Genetics, and Physiology Laboratory in Baton Rouge, LA. Pure naturally mated Russian queens are available from the Russian Bee Breeders Association members.
History In 1905 the trans-Siberian railroad was completed, opening eastern Russia to the rest of Europe. The European honeybee was imported into this area which had only been inhabited by the Asian honey bee Apis cerana, the natural host of Varroa mites. The Asian honeybee and varroa mites have co-evolved into a balanced host/parasite relationship without much harm being done. Varroa only reproduces on drone pupae in these bees, and drones are only available part of the year, so high populations of mites never build up. When the European bees encountered varroa, things were different. Varroa is able to reproduce on worker pupae which allows extremely high numbers of mites to build up. This high infestation eventually kills the colony. Beekeepers have been keeping mite populations down at great effort and expense, using miticides such as Apistan ( fluvalinate). But today, mite resistance to fluvalinate is clearly taking place, and will likely spread across the country just as rapidly as varroa did originally. Feral bees or bees managed without miticides have intense natural selection pressure, allowing only the most mite resistant colonies to survive. There are at least four resistance mechanisms that scientists have identified. They include, bees grooming mites off themselves and each other, hygienic behavior of removing infested pupae, acceleration of brood development, and suppression of mite reproduction. The ultimate goal of bee breeders is to produce bees with all these traits in a single stock of bees. It's hoped that the Russian bees will provide resistant genes that will let us take a giant step forward in the breeding effort. An earlier USDA introduction of bees from Yugoslavia did much in enhancing resistance to another serious pest, the tracheal mite.
These bees are bred to have a high degree of hygienic behavior known to be effective against diseases of the brood such as American foulbrood and chalkbrood. This trait is thought to be two behaviors acting insynergy, the uncapping of diseased cells, then the removal of the pathogen along with the pupae,effectively disrupting the disease lifecycle. Another trait, Varroa Sensitive Hygiene (VSH) is especially effective at targeting varroa mites. Dr. Spivak played a pivotal role in characterizing this behavior.
Dr. Spivak has been very active for many years in educating and promoting the use of hygienic bees to naturally combat some of the problems facing bees and beekeepers today. She has supplied seed stock to bee breeders around the country to help get a critical mass of resistant bees in the country's honeybee population. Read more about "New Direction for Minnesota Hygienic Line of Bees".
Carniolans are known for their winter hardiness and rapid spring build up.They are excellent honey producers and comb builders. They have proven to be exceptionally resistant to tracheal mites.This is our darkest colored line of bees. Our Carniolans have been crossed with VSH bees for enhanced resistance to Varroa mites.
The bee is the subspecies of the Western honey bee that has naturalized and adapted to the Kočevje (Gottschee) sub-region of Carniola (now in Slovenia), the southern part of the AustrianAlps and the northern Balkans. These bees are known as Carniolans, or "Carnies" for short, in English. At present this subspecies is the second most popular among beekeepers (after the Italian bee). It is favored among beekeepers for several reasons, not the least being its ability to defend itself successfully against insect pests while at the same time being extremely gentle in its behavior toward beekeepers. These bees are particularly adept at adjusting worker population to nectar availability. It relies on these rapid adjustments of population levels to rapidly expand worker bee populations after nectar becomes available in the spring, and, again, to rapidly cut off brood production when nectar ceases to be available in quantity. It meets periods of high nectar with high worker populations and consequently stores large quantities of honey and pollen during those periods. They are resistant to some diseases and parasites that can debilitate hives of other subspecies.
Anatomy and appearance
Carniolan honey bees are about the same size as the Italian honey bee race, but they are physically distinguished by their generally dusky brown-grey color that is relieved by stripes of a subdued lighter brown color. Their chitin is dark, but it is possible to find lighter colored or brown colored rings and dots on their bodies. They are also known as the "grey bee".
Carniolan bees are nearly as big and long as the Western European black bees, though their abdomens are much slimmer. Furthermore, the Carniolan bee has a very long tongue (6.5 to 6.7 mm, which is very well adapted for clover), a very high elbow joint and very short hair.[1]
Character and behavior
Beneficial
Considered to be gentle and non-aggressive
Can be kept in populated areas.
Sense of orientation considered better than the Italian honey bee race
Less drifting of bees from one hive to a neighboring hive
When compared to the Italian race, they are not as prone to rob honey
Able to overwinter in smaller numbers of winter bees; honey stores are conserved.
Able to quickly adapt to changes in the environment
Better for areas with long winters
Rhythm of brood production very steep. Brood rearing is reduced when available forage decreases
These golden yellow Italians are excellent for commercial pollination as well as honey production. 
