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Apicultural Research

A New Mini Syringe for Multiple Inseminations of Honey Bee Queens
by TIAGO MAURÍCIO FRANCOY1 and LIONEL SEGUI GONÇALVES2 Manuscript received for publication June 3, 2004
ABSTRACT
Instrumental insemination of queen honey bees has been very useful for genetics studies, maintenance of bee lines and genetic improvement programs. We developed a new, inexpensive mini-syringe device, capable of multiple inseminations. The syringe itself is a plastic insulin syringe, and the base and holder are made of stainless steel. The syringe tip consists of a glass capillary tube, which contains the semen, attached with silicone tubing. This insemination device is simple and inexpensive to build and maintain, and it is easy to manipulate. We initially tested the mini syringe by inseminating 12 queens; all of them survived and started to lay eggs, which originated worker larvae within a few days after the inseminations. The main advantages of this new syringe model (the Francoy-Gonçalves Mini Syringe) are the possibility of multiple inseminations, which saves time in the insemination process, and the fact that it is constructed with inexpensive, but durable material. The entire process became faster, easier and more efficient. The new mini syringe also allows us to inseminate queens with stored semen in glass capillaries, by just changing the insemination tip for another pointed capillary with stored semen.

Mackensen, 1948, 1954; Mackensen & Roberts, 1948; Laidlaw, 1949a,b). The main contribution of Mackensen was the development of the syringe that is now part of all models of inseminators. The model developed in 1948 incorporated a liquid plunger in a removable plastic tip. The liquid plunger was controlled by a diaphragm made of rubber, set in the base of the tip. This syringe is still used widely, and it allows full insemination of a single queen or a couple of queens, depending on the amount of semen in the tip. Professor Warwick E. Kerr first introduced instrumental insemination into Brazil, using the equipment developed by Laidlaw, which had the Mackensen syringe adapted to the inseminator. Later, Gonçalves & Brites (1970), while working in Kerr's lab, developed a new model of instrument whose design was based totally on screws, avoiding the utilization of "racks and pinions", therefore of easier construction. The syringe in this instrument was based on Mackensen's design.

INTRODUCTION ince its development, the instrumental insemination of honey bee queens has been used mainly for research. This technique has made possible total control of mating, the maintenance of honey bee lines and genetic markers, and the technification of selection and genetic improvement programs. Since instrumental insemination techniques were first developed, various changes have been made. The first attempt to instrumentally inseminate queens was made by Francis Huber, who painted drone semen on the vaginal orifice of queens with a brush (Huber, 1814). However, he did not succeed. Many other attempts have been made by various researchers, but without success. It was only in 1927 when L.R. Watson published his research that the technique of instrumental insemination of queens was accepted as viable. Later, one of his students, Nolan (1932, 1937) improved the technique and instruments, and these were used as a basis for the model developed by Roberts & Mackensen, H.H. Laidlaw and others, whose instruments are widely in use today (Roberts, 1947;

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Figure 1: Lateral view of the Francoy-Gonçalves Mini Syringe.

1 - Genetics Department, Medical School, University of São Paulo, 14.049-900 Ribeirão Preto, SP, BRAZIL tfrancoy@rge.fmrp.usp.br 2 - Biology Department, Faculty of Philosophy, Sciences and Letters, University of São Paulo, 14.040-901 Ribeirão Preto, SP, BRAZIL lsgoncal@usp.br Telephone number: +55 (16) 602 3154 Fax number: +55 (16) 633 6482 Corresponding author e-mail: lsgoncal@usp.br

Figure 2: Close-up view of the Francoy-Gonçalves Mini Syringe.

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American Bee Journal

August 2004 629 . and easy to manipulate (Figures 1 to 5). Figure 4: Insemination of a queen honey bee with the mini syringe. An important improvement of the instrumental insemination syringe is the Harbo syringe. also capable of multiple inseminations. It consists of an exchangeable plastic cylinder (a 1 mL insulin syringe). There is also a glass capillary with a tip that works as a syringe tip for the insemination (Figures 4-5). linked to a silicone capillary tube that is graduated in 1 µ1 (Figure 3). The development of the Francoy-Gonçalves Mini Syringe was based on the design and working principles of the Harbo syringe. It is constituted of a circular base linked to a cylindrical column that supports the Figure 3: Technical drawing of the mini syringe. This syringe is both simple and inexpensive to build and maintain. The other parts of the apparatus are a plunger and the metal support of the syringe (Figures 1-2).In 1976. which uses glass capillary tubes and allows multiple inseminations of queen bees without the need for individual semen collection for each new queen (Harbo. 1985). Gonçalves reduced the length of the Mackensen syringe to facilitate its use in Laidlaw's apparatus. Figure 5: Close-up of insemination with the mini syringe. We have developed a new mini syringe. The metal support is made of stainless steel.

(1948). Amer.S. Laidlaw. & W. The most time-consuming procedure during the normal process of insemination is the collection of semen from the drones. 47:765-768. & Brites. This new insemination device allows semen collection from many drones in succession. Econ. 89: 566-567. but durable material. CA 96073 B B B B Toll Free 888-565-8439 B B BBBBBBBBBBBBBBBBBBBBBBBBBBBBBBB Ac d e oo nc G pta ce Wooten’s Golden Queens 630 American Bee Journal Pr H od ig uc h tiv ity rly up Ea ildBu . Bee J. Laidlaw. L. Bee J. Amer. 40: 445-446. Harbo. Ent. Ent. W.S. No treatment for Tracheal Mites for 13 years. A manual for the artificial insemination of queen honey bees. and the fact that it is constructed with inexpensive. David De Jong helped correct the language. avoiding the repetition of the entire process for every new queen. 23rd. A new syringe for the artificial insemination of queen bees.C. TX 77868 BBBBBBBBBBBBBBBBBBBBBBBBBBBBBBB B B Homer Park Italian Queens B B A Family Tradition B B in Northern California B B B B B B Top Quality-All around queens B B that perform well in all areas B B B B 50 or more. brood. (1954). New observations upon bees. W. Rept. Annual Meeting pp 20-34. Multiple matings of the queen bee. MITE SOLUTION! c/o Robert B. In: Maryland State Bee Keepers Association Report. C. Ent. Econ. O. (1949b). (1814). Currently there are several ways to store semen. Hamilton Ill. Parsons 2460 Amherst Rd. storage in glass micro-capillary tubes is quite common. modified Pasteur pipette that works as a syringe tip socket. The capillary tip is fixed to a 4 cm long. (1937). Instrumental Insemination of queen bees 1. J. 39:700705. which allows for accurate control of plunger displacement. (1947). (A note on a new equipment used for the insemination of hymenoptera queens). it is also possible to inseminate queen bees with semen stored in glass capillaries by just changing the insemination tip for a capillary with stored semen and using it as a syringe tip. F. Mackensen.mini-syringe body (Figure 3). using the Mackensen syringe model or the model developed by Gonçalves. L.net/~pgold Send checks to: by Steve Tuttle. (1976). O. • Healthy Bees year around • No negative effects on queen. (1985). Improved apparatus for inseminating queen bees by the Watson method. Bee J. Navasota. Econ. 125: 197-202.com 936-825-7312 beeweaver. Amer.R.00 plus s/h $ Parsons’ Gold Apiaries G e Be ntle es Ð BUCKFAST ALL STAR BeeSMaRt RUSSIAN info@beeweaver. W.) Associate Professor Thesis. Roberts. U. where the screw plunger is located. The main advantages of the Francoy-Gonçalves insemination device are the possibility of multiple inseminations. Another advantage is the facility of sting chamber visualization during the introduction of the tip into the queen vaginal orifice. Florianópolis-SC. Development of precision instruments for artificial insemination of queen bees. 119122. pp 36-41.C. L.H. which originated worker larvae a few days after the insemination. H.Et-250. Gonçalves. August $10. Iowa. J. (Directional selection in two inbred lines of Apis mellifera L. State Apiarist.J. Jr. The glass tip has a smaller diameter than the plastic tip. Roberts. the exchangeable plastic cylinder is on the opposite end. Seleção direcional em duas linhagens endocruzadas de Apis mellifera L.75 per Queen B B Shipping extra B B B B B B B B B B 11189 Deschutes Rd.com 16481 CR 319. J. J. Nolan.J. (1932). (1948).R. Ent. Mackensen. (1927). 42:254-261. Our tests showed that the insemination of three or more queens became easier and faster than with traditional equipment. Nota sobre um novo instrumental usado na inseminação de rainhas de himenopteros.Ent.F. Genetics Department. Bee Grooming Stimulant! Herbal Bee Calmer Oil & Jelly “The Only Solution for Keeping Honey Bees Healthy!” • Grooming is the best natural mite destructor control. Ribeirão Preto. 5000 colonies that are chemical free— Yours can be too! For a free 2005 catalog email or call today! 100 Treatments-$18.S. Mackensen.Bur. Nolan. does not get into honey or wax 419-235-7037 • pgold@bright. (1949a). The plunger is directly linked to a screw. Dadant. This modified pipette is used as a support in the Gonçalves syringe holder (Gonçalves and Brites 1970) and in the Laidlaw instruments. 88:412. (1970). Proceeding of the I Brazilian Apiculture Congress. Thus. Some improvements in method and syringe design in artificial insemination of queen bees. University of São Paulo. Controlled mating in the honey bee.. Huber. Bee J. OH 45806 Bee Weaver Queens Pay!!! No treatment for Varroa Mites for 3 years. Cridersville. or workers • Usage can increase honey production • Safe.net www. J.and Plant Quar. ACKNOWLEDGMENTS The authors thank Marcela B. to which the syringe tip is tightly fixed. Econ. We tested the Francoy-Gonçalves Mini Syringe on 12 queens. The mini-syringe body consists of a metallic cylinder. which makes the visualization of the vaginal valve fold and the introduction of the capillary tube tip behind the valve fold easier.P. O. H. A syringe for artificial insemination of honeybees. Laure and João José dos Santos for technical assistance with insemination and in the apiary and Luis Augusto Magrini for the construction of the mini syringe. New instruments for artificial insemination. All of them survived and soon laid eggs. Amer.H. President of Tuttle Apiary Labs LITERATURE CITED Gonçalves. B B Palo Cedro.P. Watson . which saves time in the insemination process. pp.bright. since the glass capillary tube is too thin to be used without this complement. J. Jr.