Queen introduction is a standard practice utilized by beekeepers to introduce mated or virgin queens into queenless honey bee colonies. This procedure plays a crucial role in maintaining and managing healthy honey bee colonies. The quality of the queen bee is of utmost importance as it directly impacts the productivity and overall performance of the colony. Several environmental factors can influence the quality of queen bees. These factors include the age of the grafted larvae, the origin of the larvae, the number of young worker bees present, the availability of food in starter and finisher colonies and the successful mating of queen bees with an adequate number of drone bees. Understanding and managing these factors are essential for beekeepers to ensure the successful introduction and long-term success of queen bees in honey bee colonies [1]. Forexample, the queen may be placed at the hive entrance and simply allowed to walk in, after several puffs of cool, dense smoke have been blown into the hive entrance [2]. Another method is to spray a fine mist of light sugar syrup onto both the queen and frames of brood and worker bees taken from the brood chamber of the recipient hive [3]. The queen is then released directly on top of the frames and is sprayed again as she walks down the gap between two neighbouring frames [4]. Discrimi-nating among honeybee species is important for beekeeping and preserving honeybee bio-diversity. The honeybee species Apis mellifera L. is indeed widespread in Africa, Europe and parts of Asia, as stated by Ruttner et al. in 1978. Honeybees exhibit variations in their morphology, behavior and physiology due to their adaptation to different environmental   conditions,   as   mentioned   by   Laidlaw  and Page [5]  proposed  that  there  are  at  least  24  subspecies  of Apis  mellifera,  which   can   be   categorized   into   three   or   four evolutionary branches. This classification is based on the work of Alpatov [6]. It is worth noting that the initial attempts to classify honeybee subspecies were primarily focused on characteristics  such  as  color  and  size,  as  mentioned  by Ruttner et al. [7]. The honeybee known as Apis mellifera meda, also referred to as the Median Honey Bee, was first named by researcher in the Soviet Union near the Iranian border. This particular classification of honeybee was described and its distribution was documented in 1985 by Skorikov [8]. In Iran, Apis mellifera meda is the native honeybee species. It is primarily found in Iran and Iraq but it can also be found in southeastern Turkey and northern Syria to some extent. Despite being the native honeybee of the region, not much information is available about this particular bee species. The Italian bee, Apis mellifera ligustica, is a widely distributed and standard breed known for its specific characteristics. It is relatively small in size and has a relatively long tongue (6.3-6.6 mm). The bee's abdomen is characterized by yellow rings surrounded by black bands, with the last abdominal ring being black. The legs of the Italian bee are brown and the queen's abdomen varies in color from bright yellow to light golden yellow. Italian bees are known for their calm temperament and ability to tolerate extreme cold. They have a reduced tendency to swarm and exhibit strong defensive behavior against hive theft. The breed is valued for honey production, with workers being efficient honey gatherers. The queens of Italian bees are prolific egg layers and the colonies start raising brood early, maintaining large brood areas until autumn. Italian bees are not particularly inclined to collect propolis and are resistant to wax worms. The wax produced by Italian bee workers is white and they are suitable for honeycomb production.

The experiment was conducted in the north of the city of Karbala in the Husseiniyah/Iraq. This location was chosen due to its dense and diverse vegetation cover throughout the year, including palm trees, citrus trees, white eucalyptus, deciduous trees, willows and various vegetable plants such as beans, cucumber, squash, etc. The hives were placed near a source of running water. Additionally, a shade made of reed mat and wood was provided to protect the hives from direct sunlight. Six strong colonies of hybrid bees, specifically the local Iraqi strian Apis mellifera L., were chosen for the experiment. Each colony consisted of 10 frames. It was ensured that the colonies were similar in strength and that the hive boxes and frames were made of the same type of wood and shape. The colonies were also checked to be free of diseases and pests. The colonies were randomly divided into two groups, with each group containing three colonies. Italian bee queens (Apis mellifera ligustica) and Iranian bee queens (Apis ligustica mellifera meda) were purchased. The acquisition of these queens took place on 1/15/2023, prior to the start of the experiment. After three days, imported queens were introduced into the colonies using small transparent plastic cages designed for transporting queens. These cages provided a place to put food for the queens. Each queen was accompanied by five worker bees. After two days, all the hives were opened and the bee colonies were sprayed with a sugar solution and palm pollen dust. The colonies were periodically monitored to ensure the safety and acceptance of the queens by the worker bees. The hives were then moved to an isolated place in the Karbala desert. The colonies of Italian bees (Apis mellifera ligustica) and Iranian bees (Apis mellifera meda) were placed at a distance of twenty kilometers from each other. This separation was done to prevent interbreeding and maintain the genetic purity of the respective bee populations. A sugar solution was added to the colonies at a ratio of 1:1, For five days, the colonies were supplied with a 3-liter internal feeder. The queens were monitored administratively. After the virgin queens emerged and their physical safety was ensured, they were left for insemination. the queens were monitored to ensure successful fertilization and egg-laying. and the emergence of the male brood and the brood of the Italian and Iranian workers. The cups, were transferred to Orphaned hives. The queens from the two pure strains emerged and were fertilized. The strong and good queens were selected from the emerged queens. The selected queens were transferred to swarms or artificial divisions. The parcels were divided randomly into two groups. The swarms were equipped with modern Lancastroth hives, which are specialized frames used in beekeeping. Additional frames were added to the colonies as needed. On February 15, 2023, the colonies were balanced in areas of worker brood, pollen, honey and the colonies were similar in weight. The hives were placed on an iron hive stand and the legs of the stand were immersed in mineral oil to prevent ants from climbing into the hives. All hive boxes were painted white, which is a common practice in beekeeping. The hives were also placed in a specific orientation, facing towards the sunrise. The Italian and Iranian bee strains were moved to an agricultural area containing clover and Alfa lfa plants. The biological characteristics of the two bee strains were studied. The areas of interest (bee density, brood area, pollen area, honey area) were measured every 15 days from April 15th to July 30th. The measurement method appears to be based on a wooden wire frame divided into squares, with each square is equal to 2 ang (a unit of measurement). The areas  were  calculated  using  a  method  described  by Jeffree [9].

Produce Queens

The German-made Genter was placed in the Italian strain hive and another in the Iranian strain hive. After two days, the  queens  were  placed  inside  the  Genter  device  for 24 hours to lay eggs. The queens were taken out. On the fourth day, the eggs were hatched The hives with the diabetic solution 1-1 water and sugar before the exit of the queens were made strong and balanced partitions and the virgin queens were inserted into cages. Two days later the virgin queens were opened and monitored and after being successfully accepted by the bee operators, the queens were inseminated from the same male Italian and Iranian required strains and the hives of the two strains were placed at a distance of 20 km to ensure the production of pure strains. 7 industrial swarms were made for each strain and the best three industrial swarms were selected for each strain and transferred to clover.
 

Figure 1: Queen Breeding Device

Figure 2: A: The frame of the Italian colony and B: The framework of the Iranian colony

Figure 3: A: Queen of Iranian (Apis mellifera meda) and  B: Queen of (Apis mellifera meda)

Statistical Analysis

The experiment was designed as a factorial experiment based on the randomized complete block design (RCBD). The means of the treatment being studied were compared using the Least Significant Difference Test (L.S.D) at a significance level of 0.05. The statistical analysis was performed using the Genstat program.

Area of Brood

The results of the study indicate that there were significant effects of the bee strain, examination dates and their interaction on the closed brood area in honey bee hives.

Specifically, the Italian bee strain was found to be significantly superior in increasing the closed brood area compared to the Iranian bee strain. The Italian bee strain resulted in a closed brood area of 1825.5 ng2, while the Iranian bee strain had the least closed brood area at 901.0833 ng2. This represents an increase of 102.59% in the closed brood area when using the Italian bee strain.

Furthermore, the examination dates of the honey bee hives also had a significant effect on the closed brood area. The examination conducted on June 30th yielded the highest closed brood area of 2295.66 ng2, while the examination on July 30th resulted in the lowest closed brood area of 664.17 angstroms. This represents an increase of 245.64% in the closed brood area when comparing the highest and lowest examination dates.

The interaction between the bee strain and hive inspection dates also had a significant effect on the closed brood area. The treatment involving the Italian bee strain and the hive inspection date of June 30th produced the highest closed brood area of 3366.67 ng2, while the treatment involving the Iranian bee strain and the hive inspection date of July 30th resulted in the lowest closed brood area of 456.67 ng2. The findings suggest that the Italian bee strain, particularly when combined with early hive  inspection  dates,  promotes  larger  quantities  of eggs   and   maintains   larger   areas   of   brood   until autumn. This information aligns with previous research indicating that Italian bee strains exhibit strong brood-rearing characteristics and early initiation of brood production [10] (Table 1).

Table 1: The Effect of Strain Type and Examination Date on Brood Area

Area of Pollen

The study presented in Table 2 examined the effect of bee strain and examination dates on the area of closed pollens. The results indicated that the Italian bee strain had a significant impact on increasing the area of closed pollens compared to the Iranian bee strain. The Italian bee strain resulted in an average grain area of 1355.7 inches, while the Iranian bee strain had the lowest pollen area.

Table 2: The Effect of Strain Type and Examination Date on Pollen Area

Additionally, the examination dates also showed a significant effect on the area of closed pollens. The examination date of 7/30 performed the best, with an area of closed pollens amounting to 292.33 ang2. On the other hand, the examination date of 15/4 had the lowest area of closed pollens, measuring 208.33 ang2.Furthermore, the interaction between bee strain and hive examination dates also played a significant role in determining the pollen area. The treatment involving the Italian bee strain and the hive examination date of 15/6 resulted in the highest pollen area of 399.33 ang2. Conversely, the lowest pollen area of 125.00 ang2 was observed when the Iranian bee strain was combined with the hive examination date of 15/4.

Area of Honey

According to the results, the Italian bee strain performed significantly better in increasing the closed honey area compared to the Iranian bee strain. The Italian bees produced a closed honey area of 1162.58 ang2, while the Iranian bees produced the lowest amount at 589.58 ng2, representing an increase of 97.19%.

The examination dates of the honey bee hives also had a significant effect on the closed honey area. The inspection date of 6/30 yielded the highest closed honey area of 1528.17 ng2, outperforming all other examination dates. On the other hand, the inspection date of 4/16 resulted in the lowest closed honey area of 364.67 ang2, indicating an increase of 319.05%.
 

Furthermore, the interaction between the bee strain and hive inspection dates was found to have a significant effect on the closed honey area. The treatment involving the Italian bee strain and the hive inspection date of 6/30 produced the highest closed honey area of 1956.33 ang2. Conversely, the treatment involving the Egyptian bee strain and the inspection date of 7/30 resulted in the lowest closed honey area of 250.33 ang2 (Table 3).

Table 3: The Effect of Strain Type and Examination Date on Honey Area

Italian bees are known to have a relatively long tongue, measuring around 6.6-6.3 mm. This anatomical characteristic allows the Italian bee workers to collect nectar from deep flowers more effectively. The relationship between the bees' anatomical and phenotypic characteristics and their activity in collecting pollen and honey is highlighted.

Bee Density

The Italian bee strain was found to significantly increase bee density compared to the Iranian bee strain. The Italian bee strain resulted in a bee density of 2884.25 ng2 (units), while the Iranian bee strain had the lowest bee density of 1980.87 ng2. This indicates that the Italian bee strain was more effective in increasing bee density, with a 45.60% increase compared to the Iranian bee strain.

The examination dates had a significant impact on bee density in the beehives. The examination conducted on date 6/15 showed the highest bee density of 2992.17 ng2, outperforming all other examination dates. On the other hand, the lowest bee density of 1821.16 ang2 was observed at the examination conducted on date 7/30. This indicates a 64.30% increase in bee density from the lowest to the highest examination date (Table 4).

Table 4: The Effect of Strain Type and Examination Date on Bee Density

The interaction between bee strain and examination dates also had a significant effect on bee density. The treatment involving the Italian bee strain and examination date 6/15 resulted in the highest bee density in     the     beehives,     amounting    to    1180.00 ng2  (units).

Conversely, the treatment involving the Iranian bee strain and examination date 7/30 showed the lowest bee density.

According to the results obtained from the study, the results of the study suggest that it is possible to produce and breed pure strains of imported breeds under the climatic conditions of Karbala, Iraq, using local Iraqi breeds as the base population. The study indicates that the Italian strain out performed the Iranian breed in all biological characteristics examined.

Based on these findings, the study recommends raising the Italian breed in the climatic conditions of Karbala instead of importing and raising the Iranian breed.

1. Pérez-Sato, J.A. et al. “Improved Technique for Introducing Four-Day Old Virgin Queens to Mating Hives That Uses Artificial and Natural Queen Cells for Introduction.” Journal of Apicultural Research, vol. 46, 2007, pp. 28-33.

2. McCutcheon, D. “Queen Introduction.” Bee World, vol. 82, 2001, pp. 5-21.

3. Morse, R.A. Rearing Queen Honey Bees. Wicwas Press, Ithaca, New York, 1979.

4. Laidlaw, H.H. and R.E. Page. Queen Rearing and Bee Breeding. Wicwas Press, Cheshire, Connecticut, 1997.

5. Shawer, M.B. “Major Pollen Sources in Kafr El-Sheikh, Egypt, and the Effect of Pollen on Brood Area and Honey Yield.” Journal of Apicultural Research, vol. 1, 1991, pp. 43-46.

6. Alpatov, W.W. “Biometrical Studies on Variation and Races of the Honeybee, Apis mellifera L.” Revue de Biologie, vol. 4, 1929, pp. 1-57.

7. Ruttner, F. et al. “Die Honigbienen des Iran. Apis mellifera meda Skorikow, die Persische Biene.” Apidologie, vol. 16, 1985, pp. 241-264.

8. Skorikov, A.S. “A New Basis for the Revision of the Genus Apis.” The Review of Applied Entomology, vol. 4, 1929, pp. 249-270.

9. Jeffree, E.P. “A Shaped Wire Grid for Estimating Quantities of Brood and Pollen in Combs.” Bee World, vol. 58, no. 3, 1958, pp. 105-118.

10. Tofilski, A. “DrawWing: A Program for Numerical Description of Insect Wings.” Journal of Insect Science, vol. 4, 2004, pp. 1-5.