الفهرس 
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Abstract
This study was done in Belgium between the apiary and the laboratory of zoophysiology in the faculty of science at Gent University. The race was used for breeding program was Apis mellifera carnica.
Virgin queens were reared through two years 2010 & 2011 for two generation F1&F2 from the mother strain produced in 2008, as well as the virgin Carniolan ones using the method of Doolittle method (1915). One group of these produced virgin queens which naturally mated at Gent region and the second group was naturally mated at isolated region (Island) in Germany with the drones available in these two regions. The third group was artificially inseminated in the laboratory of zoophysiology in faculty of science at Gent University in 2010 &2011 for F1 and F2.
The groups of naturally mated queens and artificially inseminated queens were used for five studies, first: morphometrical analysis study between all colonies headed by two kind of mated queens. Second: hygienic behavior study between the same colonies headed by the previous two kinds of mated queens. Third: comparison study between the number of spermatozoae inside the spermatheca for the natural mated queens and the artificial inseminated queens in the beginning and in the end of season 2010 and 2011. Fourth: study the number of patrilines and genetic relatedness between the offspring from the mother queen and the offspring from the three daughter queens F1 by using Microsatellite DNA analysis. Fifth: comparison study between the colonies F2 productions (numbers of eggs, numbers of larvae, numbers of closed brood, the weight of honey and the area of pollen) headed by natural mated and artificial inseminated through season 2011.
The obtained results could be summarized as follow:
1- Morphometrical analysis study by using cubital index for F2 Apis mellifera carnica colonies:
This study indicated the cubital index between the ten F2 colonies headed with naturally and artificially mated Apis mellifera carnica queens. The statistical analysis showed significant difference between the ten colonies and the average for the cubital index which were 3.14, 2.66, 2.84, 2.99, 2.76, 2.91, 2.81, 2.97, 1.96 and 2.13. That is means that all these colonies were caraniolan strain because all their cubital index between the category from 2.1 to 3.3 cubital index.
2 - Hygienic behavior by using pin killer test for studying the health of ten F2 Apis mellifera carnica colonies:
The pin test was done for the ten colonies F2 headed with naturally mated queens and artificially inseminated queens. The pin test was measured the percentage of the uncapping cells, the removed cells and remained capping cells through three times, after 6 hours , after 18 hours and after 24 hours. The statistical analysis indicated that the percentage for adding the number of uncapping cells to the number of removing cells after 24 hours and the results of the ten colonies sequentially were 95.93%, 90.57%, 95.00%, 97.83%, 97.50%, 90.00%, 100%, 97.25%, 91.66% and 93.75%. The previous results indicated that the hygienic behavior was 100% in colony number 7 with artificial inseminated queen after 24 h, followed by 97.83% and 97.50% for the two colonies of 4 and 5 with natural mated queens and colony number 8 (97.25%) with artificial inseminated queen. Also the two colonies of natural mated queens number 1 and 3 recorded 95.93% and 95.00%, followed by colony number 2 (90.57%) with natural mated queen and colony number 6 (90.00%) with artificial inseminated queen.
3- Comparative study between the numbers of spermatozoae inside spermatheca of naturally mated and artificially inseminated queens through two years 2010 and 2011 under European climatic conditions:
3-1- Numbers of spermatozoae in the naturally mated queen’s spermatheca at the beginning and the end of season 2010.
The average number of spermatozoaee in the spermatheca of the dissected Carniolan queens mated naturally ”F1” in the beginning of season 2010 was 5.633 million while corresponding average number of spermatozoaee for the eight dissected queens in the end of season 2010 was 3.258 millions. Statistical analysis showed high significant differences between the two averages. It means that the available drones in Gent region not enough for mating the queens naturally in the end of season 2010.
3-2- Numbers of spermatozoae in artificial inseminated queens F1 at the beginning and at the end of season 2010.
The average number of spermatozoaee in Carniolan queens ”F1” artificially inseminated in the beginning of season 2010 was 5.908 million, compared to 3.963 million, on average, for those artificially inseminated queens in the end of season 2010. Statistical analysis showed significant differences between them. These results indicated that the artificial inseminated queens were affected by the changed and bad weather in the end of season.
3-3- Numbers of spermatozoae in the natural mated queens F2 at the beginning and at the end of season 2011.
The average number of spermatozoaee in the spermatheca of the dissected carniolan queens mated naturally ”F2” in the beginning of season 2011 was 5.609 million while corresponding average number of spermatozoaee for the eight dissected queens in the end of season 2011 was 3.181 millions. Statistical analysis showed high significant differences between the two averages. It means that the available drones in Gent region not enough for mating the queens naturally in the end of season 2011.
3-4- Numbers of spermatozoae in artificial inseminated queens F2 at the beginning and at the end of season 2011.
The average number of spermatozoaee in Carniolan queens ”F2” artificially inseminated in the beginning of season 2011 was 5.535 million, compared to 4.106 million, on average, for those artificially inseminated queens in the end of season 2011. Statistical analysis showed significant differences between them. These results indicated that the artificial inseminated queens were affected by the changed and bad weather in the end of season.
3-5- Comparison between the number of spermatozoae in the natural mated queens and the artificial inseminated queens at the beginning of season 2010.
Statistical analysis proved that there is no significant difference between the queens ”F1” mated naturally with average 5.633 millions spermatozoae in the beginning of season 2010 and the queens artificially inseminated with the average of 5.908 spermatozoae in the beginning of the season. It is mean that artificial insemination has a good result like natural mating and also that may due to the ability of the queen in the beginning of the season to store enough numbers of spermatozoae.
3-6- Comparison between the number of spermatozoae in natural mated queens and artificial inseminated queens at the end of season 2010
Data of the present study stated that the stored numbers of spermatozoae inside the spermatheca were decreased after the mating naturally at the end of season and counted 3.285 million compared with 3.963 million after artificial insemination although each queen injected in the common oviduct with eight microliter semen fluid. It may be explained for the natural mating by the decreasing of drones at the end of season, while in the artificial inseminated queens it may be due to the decreasing of the queen ability to store semen.
3-7- Comparison between the number of spermatozoae in the natural mated queens and artificial inseminated queens in the beginning of season 2011.
Statistical analysis showed that there is no significant difference between the queens ”F2” mated naturally with average 5.610 millions in the beginning of season 2011 and the queens artificially inseminated with average 5.534 in the beginning of season under the same conditions. It is mean that artificial insemination has a good result like natural mated queens and also it may be due to the statues of the queen in the beginning of the season to save enough numbers of spermatozoae.
3-8- Comparison between the number of spermatozoae in the natural mated queens and artificial mated queens in the end of season 2011.
Statistical analysis illustrated that there was significant difference between the queens ”F2” mated naturally and stored average of 3.181 millions in the end of season 2011 and the queens artificially inseminated which stored average of 4.110 in the end of season under the same conditions. It is mean that artificial insemination has the same effect with the weather changed and the bad conditions like natural mated queens. Although, in the end of the season the average number of spermatozoae in artificial inseminated queens was higher than the average number of spermatozoae in natural mated queens.
4- The number of patrilines and genetic relatedness between the offspring from the mother queen and the offspring from F1 queens by using Microsatellite DNA analysis:
The five primers ACOO6, A76, A107, A43 and B124 were used in Microsatellite and the statistical analysis showed significant difference between them. Also it is found diversity between the mother offspring and her three daughter offspring.
The first primer ACOO6 showed a significant difference between each group and the average of molecular weight of the mother offspring and her three daughter offspring which were 170, 172, 164.4, 179.8 bp. The DNA weight showed a relationship between the mother offspring and the first daughter offspring through primer ACOO6.
The second used primer was A76 and the statistical analysis proved that there was significant difference between each group and the average of molecular weight of the mother offspring and her three daughters offspring which were 314, 238.4, 300, 330 bp. The result of the DNA weight indicated that there was diversity between the mother offspring and the three daughter offspring through primer A76. It is mean that queens were mated with many drones and the offspring has many patrilines.
The third primer was A107 and the statistical analysis indicated that there was significant difference between each group and the average of molecular weight for the mother offspring and her three daughter offspring which were 191.4, 188.8, 195.8, 190.8 bp. The DNA weight indicated that there was diversity between the mother offspring and the three daughter offspring through primer A107. It is mean that queens mated with many drones and the offspring has many patrilines. The molecular weight for the mother offspring 191.4 bp was closed to the molecular weight for the third daughter offspring 190.8 bp. It is mean that there was a relationship between the mother and her third daughter offspring.
The fourth primer was A43 and the statistical analysis indicated that there was significant difference between each group and the average of molecular weight for the mother offspring and her three daughter offspring which were 145.6, 143, 147.2 and 150.2 bp. The DNA weight indicated that there was diversity between the mother offspring and the three daughter offspring through primer A43. It is mean that queens mated with many drones and the offspring has many patrilines.