NEW METOD IN ERADICATING AMERICAN FOULBROOD (AFB) USED IN A APIARY WITH 56 HONEY BEE COLONES (APIS MELLIFERA CARNICA) IN RIMSKI SANCEVI NEAR NOVI SAD – AP VOJVODINA – SERBIJA – YUGOSLAVIA
AUTHOR: NAUM BANDZOV, Mr. of Apiculture, Fruškogorska 25 Maradik AP Vojvodina – Serbija -Yugoslavia Tel: **381 22 506 676 E-mail: email@example.com
ASSISTANS: Dr. D. Djermanovic, Dipl. vet. D. Kolaric – Vet. Institute, Zemun
Mr. vet. D. Suvakov, Dipl. vet. N. Plavsa, Dr. vet. M. Mrdjen – Vet. Institut Novi Sad
Dipl. vet. M. Maljkovic Vet. Inspector of Province Vojvodina – Novi Sad
Beekeeper Z. Lovrenovic, D.Pantic, M. Mitrovic. BeekeeperI. Boros – apiary owner
The Fourth amended edition of the „Veterunary Manual“ from 1989 which is nowadays valid as Yugoslavian and Macedonian veterinary doctrine, in its Chapter XI discusses bee diseases. On pages 701-702 it recommends eradication of AFB by two methods:
1. Eradication of the affeered colony, and
2. Cure of infested communuty by treatment and search selection
We improved the seconds method relating to continual beehives with mobile honey combs. The treatment was divided into two phases: procedure prior to drug application consisting of the following: frominfested and ill colonies we extracted all honey and took out frames with infested larvae and pleaced them into special ISOLATORS (inovation to the procedure) untill all young and healty bees came out, which were later on returned to weaker colonies for improvement of generation structure and higyenic behavior. When all young-healthy bees come out we took the affected sombs from isolators, cut them out the frames, burned and burried them deep into the ground. The hives of affected colinies were replaced with new desinfected ones, frames of foundation. Affected colony hives were disinfected by scrubbing and flame.
With this new tehnologie – biologie genetic solution we were capable of eradicating the infestation on a large apiary in only one to three days and place it under control. Later on we initiated the second phase: DRUG APPLICATION in orderto prevent further infestation of the youngest 24 hours old larvae which are mostly affected, in 44,1% of cases, while 24-48 hours old larvae are affected only in 4,8%, and 2-4 days old larvae became completely resistant to AFB infection. We applied Vesemycin AT, 75 mg of active substance oxytetracycline in 1 l of sugar syrup 1:1. The drug was applied every other day in two ways, depending on the strenght of community: a) in stronger communities we added 200-500 ml into the nursery, and b) in weaker colonies which lost a bee generation due to severe infection were sprayed the 100-200 ml of medicinal syrup over wax frame producing bees. The treatment with the medicinal syrup lasted 15 days.
Foto 1, 1a. Foto 2. Foto 3.
Foto 4. Foto 5. Foto 6, 6a.
Foto 7 Foto 7a
Foto 1, 1a. Apiary in Rimski Sancevi when infection was registered.
Foto 2. Taking samples for microbiological analiisys.
Foto 3. Frames whith no healthy brood was been send for melting and desinfection.
Foto 4. Aplication of medicin efor smail colonies.
Foto 5. One of INSULATORS where young bees has emerget and later hes been gaven to small colonies for improvement of their generation structure.
Foto 6, 6a. TRANSFUZION with young bees from INSULATORS to smail colonies.
Foto 7. 7a. Samples of old dark and young light combs with normal healthy brood after sanation.
The American foulbrood (AFB) has become a world problem due to not defined legal regulation and heterogeneous veterinary AFB eradicating of stock diseases in SR Yugoslavia and R Macedonia in aim to cure of apiaries with AFB diagnose all infected colonies must be eradicating, namely burned and buried into the ground, together with immobile comb beehives, lasted beehives with mobile combs including bees and combs.
The Veterinary Manual from 1989, which is valid for SR Yugoslavia and R. Macedonia foresees in its Chapter XI on the pages 701-702 (Authors: Dr. Nikola Fijan & Dr Djuro Sulimanovic) cure possibility of infected colonies by healing and search selection. On the base of this possibility we have introduced the new TECHNOLOGICAL – BIOLOGICAL – GENETIC METHOD.
According to Park investigation (1936-1937) AFB killed brood are removed from brood combs and the hygienic behaviour of the bees. This hyginic behaviour has been confirmed by S. Taber (1982); Kulincevic J. & Rothenbuler W.C. (1982); Newton D. & Ostariewski N. (1985) on Apis Melifera ligustica and Bandzov N. (1990) on Apis Melifera var. carnica from the region of R. Macedonia – surroundigs if Skopje.
It has been also found that larvae are not susceptible to the vegetative stage of the pathogen in recently dead larvae (Tarr 1937) and the quickly removal of diseased larvae from the brood nest before sporulation helps to control AFB (Woodrow & Holst 1942).
It is known that there are, more or less, AFB – resestent bee colonies (Rothenbuhler 1967; Kulincevic & Rothenbuhler 1982; Bandzov 1990). On the basis of this characteristic more AFB resistant bees can be produced by selection. Woodrow (1942) proved, that larvae of different ages exhibit differences in ability to rasist Bacillus larvae. The effectiveness of the bee hygienic behaviour is increased during a nectar flow (Mamot & Rothenbuhler 1971). Adding of the pure sugar syrup increases the hygienic behaviour (Thomson 1964; Murer 1964).
Sturtevant and Ravel (1953) found that bees separate up to 79% Bacillus larvae spored by honey valve before they place the syrup into the combs. Thompson & Rothenbuhler 1957, proved the hygienic behaviour of the spore removal from the food for larvae, also by honey valve. The investagations of the infection with a number of AFB spored and vegetative cells, made by Tarr (1937), AFB spores in the extracted honey during 6 years, made by Hansen & Rasmunsen (1986) as well as report from Bandzov N. at the International Symposium (Zagreb 1986), point contrary to the former presumption at the fatal presence of the small quantity of spores in a bee community and AFB phenomenon.
All previous knowledges about AFB and its eradication have encouraged us for two years investagatins (1996 – 1996) in Rimski Sancevi and Maradik to apply the new AFB eradicating method, in the first place, the TECHNOLOGICAL – BIOLOGICAL – GENETIC METHOD with introduction of new original solutions by forming insulators with infested frames, coming out of not infected young bees, returning of them for improvement of structure and hygienic behaviour. It can be said in biological language we undertook a YOUNG BEE TRANSFUSION. It is known that BLOOD TRANSFUSION is the best medicament in the humane medicine for affected people.
MATERIALS AND METHODS
The investagations have been made near Novi Sad, in Rimski Sancevi on the apiary of 56 honey bee colonies, owner Boros Istvan & Marica. Our work lasted from July 8, 1995 till September 3, 1996, as we made the last microbiological analysis. The disease have been found by the beekeeper Lovrenovic Z. on June 27, 1995 (the owner was absent for 6 months in Canada). The beehives are Lankstrot – Rut type (LR) with 1-3 (extesions) and half shoulders. On August 7, 1995 we inspected all of 56 bee colnies and found the AFB disease by clinical examination in 52 bee colonies with different infection range from 1-16 frames. Only in four honey bee colony we didn’t found any infection. The authorhas microscopial proved the presence of Bacillus larvae in his private labaratory in Maradik, Fruskogorska 25. We have informed the Veterinary Institute in Novi Sad about AFB infected apiary immediately and on July 13, 1995 a commission came out there which found AFB by clinical examinations too. They took two comb samples from the brood of LR frame 43 x 20 from the beehives Nr 35 and 43 as well as bees from the beehive Nr 44. The commission made a protocol Nr 9 on July 13, 1995. The examation results have been delivered to the apiary owner in Province Veterinary Inspection in Novi Sad.
We cam up to the cure during the period from July 14-17, 1995. We extracted all surplus of the honey, placed in the beehives (130 kg). The treatment was devided into two phases. The first cure phase foresees the TECHNOLOGICAL – BOILOGICAL METHOD. From all of 52 infected bee colonies we took out 580 AFB infected frames. From these number, healty larvae and pupae couldn’t be found in 330 frames, where dead larvae with AFB were present. We sent them forwax melting and frame desinfection. The frame desinfection was made by mechanical scraping by a beekeeper knife, followed by flame desinfection. Then we placed comb foundation into the desinfected frames.
From the remaining 250 frames, where also were present living and not infected larvae and pupae together with existing bees on the frame without quinbee, we formed 5 insulators with 50 frames each in the apiary. This is a inovation compared to the older method „AFB ERADICATING IN LR BEEHIVE BY HANGING OVER METHOD“ (Zagreb 1986) published in the Magazine BEEKEEPER 10/86: pages 305-306.
Due to strong varro-infection we placed 5 laths soaked with fluvalinat to release them. The insulators have been held for 21 days, untill all young and healty bees came out, which were added to the weaker colonies step by step for improvement of generation structure, which has been lost due to strong AFB infection as well as of hygienic beehaviour. Then we sent these frames for wax melting, desinfection and comb foundation mounting too.
The frames of the foundation and LR beehive extensins were replased with desinfected elements. The frames from which honey was extracted (108) and frames which were not AFB infected and broodfree were left together with bees in „new“ desinfected beehives (foundations and extensions). We added so many frames to them es required for existing bees. When the frame number was insufficient we placed existing desinfected frames with the comb foundation.
After cure we devided all bee colonies from which were removed the infected frames (52) into three groups. On July 17, 1995 they got new indications:
I group WEAK (12) with 1-3 brood frames each and min. 3-5 bee streets. New indication: L-12.
II group MIDDLE (25) with 4-5 brood frames each and min. 5-7 bee streets. New indication: 13-37.
III group STRONG (15) with 7-9 brood frames each and min. 8-10 bee streets. New indication: 38-52.
Four beehives, not infected, have got the indications 53-56.
This TECHONOLGICAL – BIOLOGICAL METHOD realised in the subject bee community (social organism) can be called in veterinary medical dictionary SURGICAL OPERATION OF REMOVING DISEASED COMB PART (TISSUE).
The disaese in this large apiary was kept down in 3 days and placed under control. So we were capable to go on with successful rehabilitation in the second phase – drug aplication in 15 following treatments in order to prevent further infection of the youngest 24 hours old larvae, which are mostly affected in 44,1% of cases. The 24-48 hours old larvae are affected in 4,8%, and the over 48 hours old larvae became resistent to the infection (Woodrow 1942). We prepared the drug antibiotic VESEMYCIN AT (it is in the examining phase now) with the active substance oxytetracycline (we applied 75 mg of active substance oxytetracycline, dissolved in sugar syrup 1:1). We prepared the medicinal syrup in the quantity, necessary for a day. The medicinal syrup was applied in our project in two ways:
a) by adding into the nursery 200-500 ml for stronger communities, which have not lost the generation structure, every day in following 30 days (15 application with drug and 15 additional nursing with normal syrup);
b) by sprayng of combs and bees 100-200 ml for weaker colonies, which have lost their generation structure due to strong infection. To these colonies we added young bees from the insulators and so we improved slowely the generation structure of the subject colinies to be able to take the medicinal syrup from the nursery unaided.
After 30 days, the apiary could be considered as completely healthy, so we stopped drug application. No colonies have received any half micron of drugs for preventive till now. The author of project doesn’t accept preventive for healthy colonies, as Dr. Wilson, the author of GREASY CAKE, says in his doctoral dissertation „giving to the healthy colonies is forbidden“. As a pharmacist and i am competent to say that „every drug, especially antibiotic, given to a healthy organism, means a toxin“ (Pharmacopoeia).
From 4 not infected bee colonies with the indications 53, 54, 55, 56 we took materials for production of 80 queen-bees on July 20, 1995. This material was placed in the authors apiary in Maradik, 30 km far from the experimental apiary, for breeding and fertilasation, because the author produces queen-bees which are hygienic behaved, since 1986. As we begain with the preparations for winter conditions on August 15, 1995, we replaced the weaker queen-bees with 25 new fertilized. 50 bee colonies and 6 nucleuses were prepared for winter conditions in the period October 6-8, 1995. In 1996 we replaced remaining 31 queen-bees with young bees, which demonstrated excellent hygienic behaviour. 60 bee communities were prepared for winter conditions.
R E S U L T S
The labaratory eximinations results on the samples. taken on July 13, 1995, were finished oh July 14, 1995. The report No 2281 is as follows: AFB – Pastis apium – POSITIVE; Varoa Jacobsoni – POSITIVE; Ascospera apis – POSITIVE; Nosema apis – POSITIVE.
During the first cure phase in the period July 14-17, 1995, 52 bee colonies received medicinal syrup dor two times. The nectar flow was good and daily bringing amounted 600-1200 g, in middle colonies 800g. Besides existing frames (557) we placed 210 frames with comb foundation in I phase which were very quickly built and breed by queen-bees with eggs.
The second cure phase lasted from July 19, 1995 till August 12, 1995 and the bee colonies (52) were treated with medicinal syrup 13 times.
The veterinary province inspector came two times in the II cure phase to the apiary, on July 28 , 1995 and August 11, 1995 and inspected 26 bee colonies according to his finding. Non of them demonstrated any AFB symptoms. During the second cure phase we added 180 frames with comb foundation. The author and his assistants completed 13 control inspections and no AFB symptoms were found.
As we begain with the preparatios for winter conditions on August 15, 1995, we replaced 25 weakest queen-bees.
Three months after first AFB diagnosis on September 27, 1995 the researcher of Veterinary Institute – Novi Sad tool 5 new comb samples with the brood 10 x 10 according to his choice, from the beehives No. 5, 13, 18, 35 and 38 for microbiological diagnostic.
Besides subject samples he made clinical inspection of 10 bee colonies more and didn’t notice any AFB symptoms. He also made the protokol No. 11 on September 27, 1995.
The results of the laboratory microbilogical examinations were finished on October 4, 1995 under No. 3006/113. The report was: Varoa jacobsoni – POSITIVE in the sample No. 4; Pestis apium (AFB) – NEGATIVE in all of five samples; comb sowed culture – NEGATIVE.
In the period October 6-10, 1995 we prepared for winter condition 50 bee colonies and 6 nuicleuses. 48 colonies were wintered. The loss 8 bee colonies was normal for us, because they starved. Detail inspection was made on April 2, 1996. No one nee communiti demonstated AFB symptoms. The apiary gave 800 kg honey, 80 queen-bees, 20 swarms, 800 frames with comb foundations were built and 60 bee colonies were prepared for winter conditions.
On August 26, 1996 the researcher of Veterinary Institute – Novi Sad inspected 15 bee colonies at our request, according to shes choice, and no AFB symptoms were found in clinical examinations.
Shee tool samples of combs with brood from 5 bee colonies indicated 3, 8, 21, 27, 35, for laboratory analysis – diagnostic. She also made the protocol No. 1/96 on August 26, 1996.
The examination results No. 3206/213-217 of September 3, 1996 indicated in all of five samples AFB-pestis apium – NEGATIVE; sowed culture – NEGATIVE too.
D I S C U S S I O N
The two years investigations which lasted from July 8, 1995 till September 3, 1996 indicated that AFB can be eradicated successfully by combinated TECHNOLOGICAL – BIOLOGICAL – GENETIC method and by application of antibiotic Oxytetracycline in very small doses and in a very short time period.
Although the AFB eradicating in SR Yugoslavia and R. Macedonia is officially within the competency of the Veterinary services, it is known that beekeepers apply antibiotics and another drugs according to their finding, often wrong drugs and in a wrong way in undefined doses. Therefore this AFB eradicating by the combined technological – biological – genetic method (the author of this project has got very good results with this method only) and by possible therapy with oxytetracycline, presents a very important inovation compared with previous methods, by selection, overhanging; the worst is the actual legal regulation – „DESTROY AND BURN UP“
All pervious scientific investigations and experiences in the world as well as this newest AFB eradicating method, proved many times throughout the world cannot see the light for a wider application in SR. Yugoslavia and R. Macedonia due to certain administration and apiculture officials and regulations.
We hope that it is similar in other regions. From this reasons I am asking APIMONDIA to take in considering the possibility of introduction of a NEW UNIFIED WORLD VETERINARY DOCTRINE IN AFB ERADICATING AND DISCONTINUING WITH PREVIOUS OUT OF DATE MENTALITY OF 20 CENTURY „DESTROY AND BURN UP“ SO WE CAN ENTER INTO 21 CENTURY WITH A NEW MENTALITY „SAVE? PRESERVE AND GET PROFIT“.