Relationship between larvae age and characteristics of queens (2003)

 Relationship between larvae age and characteristics of queen honeybees (Apis mellifera L.) after single and double grafting (2003)

B. Emsen, A. Dodologlu
Department of Animal Science,
Ataturk University 
25240 Erzurum-Turkey

Abstract
Effect of grafting methods on characteristics of queen honeybees
Ninety mating colonies were used. Single and double grafting applied for A1 and A2 grafting groups, respectively and 1, 2 and 3 days old larvae were grafted in each group. The average pre-oviposition periods were 12.41±0.028 and 13.25±0.040 days; the average queen weights at emergence were 181.8±0.43 and 172.7±0.61 mg; the average diameters of spermatheca were 0.97±0.0001 and 0.98±0.0010 mm and average numbers of spermatozoa were 4.500.000±0.09 for the queen bees of A1 and A2 groups, respectively. Queen weight at emergency, pre-oviposition period, diameter of sperm theca and the number of spermatozoa were not different within grafting methods. Larvae age had significantly (p<0.01) effect on pre-oviposition period, diameter of sperm theca, the number of spermatozoa, and (p<0.05) queen weight at emergency. A positive important (p<0.05) correlation was observed between weights at emergency and diameter of spermatheca (r= 0.264), the number of spermatozoa (r = 0.508). This relationship was negative (p<0.05) between ovi-position period and weights at emergency (r = -0.238), the number of spermatozoid (r = - 0.367) while an important (p< 0.05) positive correlation (r = 0.365) was found between diameter of spermatheca and the number of spermatozoa.

Key Words
Grafting, larvae age, queen bee rearing

Corresponding author
Ziraat Fakultesi Zootekni
Bolumu 25240 Erzurum/TURKEY
Tel.: +90 442 23119881.

Introduction
Productivity of a colony depends on some environmental factors like climate and type of plants, queen bee, colony performance, bee diseases, genotype and beekeeping techniques. Turkey is quite available as means environmental factors for beekeeping. Colony productivity mostly is related with the factors belong to colony. The most important of those factors is queen bee [1].

Queen bee is the mother of all other individuals in the colony. The queen is the breeding depository of all of the inherited characteristics of the colony acquired through her progenitors and through the acquisition of the sperms from males at the time of mating. Thus, the queen is responsible for the color of the bees, their industry, degree of gentleness, resistance to disease, swarming tendencies, longevity, comb building propensities, and for many other colony and individual attributes [2].*

Productivity of queen bee depends on her age, breed, breeding term, weight in emergency, age of larvae and grafting methods, number of ovariol, diameter of spermatheca, number of spermatozoa in spermaethaca, and if she has anatomical disorder or not. [3, 4].

Queen bee, average 4-5 years lifetime, maintain its productivity just only first 1-2 years. Secretion of pheromones, the effects of the pheromones on the colony individuals and efficiency of laying eggs is decrease in lately age of queen. Therefore, queen bee should be replaced in every year in stable bee breeding or once in two years in traveling bee breeding. [ 4].

The objective of the current study was to determine the queen bee rearing techniques, accurate measurement used in selection and the correlation among them by using different aged larvae with single and double grafting methods to offer the best queen bee rearing technique for bee breeders.

Materials and Methods
Two feeding colonies and 90 mating colonies were used at the Agricultural Research and Application Farm in Erzurum. Doolittle grafting method was used for queen bee rearing. Two queenless breeder colonies were used for each grafting group instead of using starter or starter-finisher colony. One grafting comb with three units queen cell cups was given to each breeder colony. Forty five larvaes in group one (A1) were single grafted, 45 larvae in group two (A2) were double grafted.1, 2 and 3 day old larvaes were grafted in both grafting groups (A1, A2). Wooden cell cups were used as queen cell cup. The queen cells cups kept in feeder colony until the cups be completely closed were placed in individual plastic vials and put in an incubator set at 33.0 ± 0.05 C and 60-65 % RH [5,6, 7]. The incubator was checked every 4-5 hours for newly emerged queens. Shortly after queens emerged from her cell were weight, marked and numbered and were placed in mating colonies (nuclei) in nursery cages. Virgin queens were permitted to mate after 24 hour placed in the nucleus.

Pre-oviposition period was recorded by controlling of nucleus properly. To examine relationships between queen bees survivial and the number of sperm in the spermatheca of queen bees. Mated queens were took place in a laboratory to measure diameter of spermatheca and the number of spermatozoa. Data were analyzed by analysis of variance and means were repeated by LSD multiple range test. [8 ].

Results
The results of queen weight at emergency, pre-oviposition period, diameter of sperm theca and the number of spermatozoa were given in Table 1, Table 2, Table 3, and Table 4. There were no differences pre-ovi-position periods in queens reared by single and double grafting methods [9].

grafting 1
Table 1. The results of pre-oviposition period

The averages of ovi-position period were found as 12.09±0.054, 12.73 ± 0.053 and 13.67 ± 0.045-d in-groups of 1, 2 and 3-day-old larvae. The pre-oviposition period of 3-day-old larvae was significantly (p<0.01) higher than the results of 1 and 2 day larvae. In the result of LSD test, queen bee reared from 3-day-old larvae double grafted lay egg later than all others groups.

Discussion
In present study, results of max and min pre oviposition period, 16d and 9 d, respectively, similar to findings obtained by different researchers Gul and Kaftanoglu [10], Genc [7] and, Dodologlu and Genc [11]. In queen bee rearing, virgin queen is required to start laying egg as short as due to use season more effectively and extend period of using of queen bee in brood. Hence, preferred single grafting and using 1 and/or 2 day old lavae will be more authentic for abbreviating pre-ovipsition period. Grafting method and interaction grafting method x larvae age had no effect on emergency weight (mg) of queen bee but affection of larvae age on the same characteristic was found important (p<0.05). Emergency weight (mg) of queen bee was found between from 114 mg to 238 mg in group A1, from 128 mg to 207 mg in group A2.
a, b Means with no common superscript in a column within a parameter differ (P< 0.05).


Table2. Queen weights at emergency

Queen bees reared by 1 and 2 day old larvae had higher (p<0.05) emergency weight than reared by 3 old day larvae. Woyke [12] reported that queen bees grafted from one and two day old larvae and weighed at emergence ranged in weight between 0.156-0.201g. Queen weights at emergency obtained from this present research were similar to that reported by Rawash, at all., [13] and Nelson and Gary [14]. It can be inferred from these result that weight at emergency is creditable criteria be used for selection of virgin queens. Queen bee rearer is suggested by the finding from the present study to use young larvae and single grafting instead of double grafting, causing waste time, raise labor and lose larvae.

In comparison of treatment, there was also a significant effect of grafting methods for diameter of spermatheca and the number of number of spermatozoa, but larvae age had (p<0.01). Diameter of spermatheca and the number of queen reared by 3 day old was lower (p<0.05) than the queens reared 1 and 2 day old larvae. The results of diameter spermatheca were lower than findings reported by Woyke [12] and Kaftanoglu [15], but similar to Dodologlu [6]. Diameter of spermatheca has big importance in bee keeping. 0-24 h old larvae must be used in double grafting to obtain high quality queen bee for convenient diameter of spermatheca . Also, it is concluded that double grafting is not necessary but 1 and 2 day old larvae should be performed for queen bee with higher spermatozoid stock. In addition to these results, a positive important (p<0.05) correlation was observed between weights at emergency and diameter of spermatheca (r= 0.264), the number of spermatozoa (r = 0.508). This relationship was negative (p<0.05) between ovi-position period and weights at emergency (r = -0.238), the number of spermatozoid (r = - 0.367) while a important (p< 0.05) positive correlation (r = 0.365) was found between diameter of spermatheca and the number of spermatozoa. According to these findings, queen that it’s weight at emergency is high start lay egg earlier and queen that it’s pre-oviposition lately occurred has less spermatozoa. Therefore, getting shorter of pre-oviposition period is important to make queen has more spermatozoid in it’s spermatheca.
a, b, c Means with no common superscript in a column within a parameter differ (P< 0.05).

grafting 3
Table 3. The average number of spermatozoa and spermatheca diameter of queen bees

Acknowledgements
The authors grateful to Ataturk University for supplying a great working environment and, also to members of Rothenbuhler Honey Bee Lab to scientific review for this study .

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