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STUDIES ON ROYAL JELLY PRODUCTION IN HONEYBEE COLONIES

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STUDIES ON ROYAL JELLY PRODUCTION IN
HONEYBEE COLONIES

By

HATEM AHMED SHARAF EL-DIN


B.Sc. Agric. Sci. (Plant Protection), Fac. Agric., Cairo Univ., Egypt, 2006

THESIS
Submitted in Partial Fulfillment of the
Requirements for the Degree of

MASTER OF SCIENCE

In

Agricultural Sciences
(Economic Entomology)

Department of Economic Entomology and Pesticides


Faculty of Agriculture
Cairo University
EGYPT

2010
APPROVAL SHEET

STUDIES ON ROYAL JELLY PRODUCTION IN


HONEYBEE COLONIES

M. Sc. Thesis
In
Agric. Sci. (Economic Entomology)

By

HATEM AHMED SHARAF EL-DIN


B.Sc. Agric. Sci. (Plant Protection), Fac. Agric., Cairo Univ., Egypt, 2006

Approval Committee

Dr. MOHAMMAD BAHGAT SHAWER ………………………


Professor of Economic Entomology, Fac. Agric., Kafr El-Sheikh University

Dr. MOHAMAD ATTIA EWIES …………………………………………….


Professor of Economic Entomology, Fac. Agric., Cairo University

Dr. MOHAMMAD ABD AL-WAHHAB ABD AL-FATTAH…


Professor of Economic Entomology, Fac. Agric., Cairo University

Dr. MAHMOUD EL-SAYED NOUR ………………………………..


Professor of Economic Entomology, Fac. Agric., Cairo University.

Date: / /2010
SUPERVISION SHEET

STUDIES ON ROYAL JELLY PRODUCTION IN


HONEYBEE COLONIES

M. Sc. Thesis

In
Agric. Sci. (Economic Entomology)

By

HATEM AHMED SHARAF EL-DIN


B.Sc. Agric. Sci. (Plant Protection), Fac. Agric., Cairo Univ., Egypt, 2006

SUPERVISION COMMITTEE

Dr. MAHMOUD EL-SAYED NOUR


Professor of Economic Entomology, Fac. Agric., Cairo University

Dr. MOHAMMAD ABD AL-WAHHAB ABD EL-FATTAH


Professor of Economic Entomology, Fac. Agric., Cairo University
Name of Candidate: Hatem Ahmed Sharaf El-Din Degree: M.Sc.
Title of Thesis: Studies on Royal Jelly Production in Honeybee Colonies
Supervisors: Dr. Mahmoud El-Sayed Nour,
Dr. Mohammad Abd Al-Wahhab Abd Al-Fattah.
Department: Economic Entomology and Pesticides
Branch: Economic Entomology Approval: / / 2010
ABSTRACT
This work was carried out in the apiary of the Agricultural Experimental Station,
Faculty of Agriculture, Cairo University, Giza Governorate.
This study aimed to evaluate some factors affecting royal jelly production in
honeybee colonies as well as determination of amino and fatty acids percentages in
some samples of the produced royal jelly.
Experiments of the firt aim were separated into colonies grafted with non related
larvae and colonies grafted with related grafted larvae.
Experiments of colonies grafted with non related larvae were conducted using
Egyptian race and Carniolian hybrid queenless production colonies. Results of Egyptian
race colonies showed marked reduction in both mean percentage of acceptance and
amount of harvested royal jelly (gm) when compared with those of Carniolian hybrid
ones.
Experiments of colonies grafted with related grafted larvae were carried out using
Egyptian race, Italian and Carniolian hybrids queenless production colonies. Results
indicated that the Carniolian hybrid was the best for royal jelly production as the
hieghest percentage of acceptance and amount of harvested royal jelly (gm) were
achieved when compared with others. Italian hybrid gave higher acceptance percentage
than Egyptian race but amount (gm) of harvested royal jelly was almost the same with
an increase of 0.02 for the Egyptian hybrid.
In respect of amino and fatty acid percentages, the percentages of essential amino
acids had their maximum values in the Egyptian race samples, followed consecutively
by Italian hybrid and Carniolian hybrid samples, with an exception of Lysine acid,
where Carniolian hybrid samples gave the highest amount of it, followed by Italian
hybrid then the Egyptian race. The maximum values for non-essential amino acids
occurred in the Egyptian race samples whereas the second and third ranks oscillated
between Italian hybrid and Carniolian hybrid samples.
The maximum values of fatty acids were recorded in the Carniolian hybrid
samples, whereas the second and third ranks fluctuated between Italian hybrid and
Egyptian race samples except for both of 10-hydroxy-2-decanoic acid (10HDA)
samples and Cis,cis-9,12- Octadecadienoic acid. Italian hybrid had the highest amount
of 10HDA, followed by the Egyptian race then Carniolian hybrid samples. On the other
hand, Cis,cis-9,12- Octadecadienoic acid gave the highest value in the Egyptian race
samples then Carniolian and Italian hybrids, respectively.
Key words: Honeybee, colonies, royal jelly, grafted larvae, hybrid, race, amino acids,
fatty acids
DEDICATION

I dedicate this work to whom my heart felt


thanks; to my wife and my daughter for their
patience and help, as well as to my parents for all the
support they lovely offered along the period of my
post graduation.
ACKNOWLEDGEMENT

Firstly, and for more of all, thanks to "ALLAH" who gave


me the strength patience and ability to complete this work.
I wish to express my sincere thanks, deepest gratitude and
appreciation to Dr. Mahmoud El-Sayed Nour and Dr.
Mohammad Abd Al-Wahab Abd Al-Fattah, Professors of
Economic Entomology, Faculty of Agriculture, Cairo University
for suggesting the problem, fruitful supervision, useful advice and
their useful guidance through the course of study and revision the
manuscript of this thesis.
Grateful appreciation is also extended to Faculty of
Agriculture Research Park and all staff members in the Apiary
yard of Experimental Station, Faculty of Agriculture, Cairo
University.
CONTENTS

Page
1INTRODUCTION................................................................................. 1
REVIEW OF LITERATURE....................................…............... 4
1. Methods of production......…........................................................ 4
2. Effect of season on royal jelly production.................. 17
3. Effect of genetic origin on royal jelly production.. 23
4. Effect of feeding on the produced royal jelly …… 25
5. Development of hypopharyngeal glands………...…... 33
6. Chemical analysis of royal jelly ………………………… 36
3MATERIALS AND METHODS…………….…....................... 44
4RESULTS AND DISCUSSION…………………....................... 56
1. Evaluation of some factors affecting royal jelly
production in honeybee colonies………………………...… 56
a. Non related grafted larvae………………………………… 56
1. Egyptian hybrid grafted with different genetic
origin larvae throughout the period extended from
22/3/2008 to 31/3/2008…………………………......................... 56
a. Acceptance percentage of grafted larvae………… 56
b. Amount of harvested royal jelly…… 63
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly……………….. 65
2. Carniolian hybrid grafted with Carniolian race
larvae throughout the period from 21/4/2008 to
30/4/2008…………………………………………………………… 68
a. Acceptance percentage of grafted larvae…………….. 68
b. Amount of harvested royal jelly………………………... 76
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly……………….. 76
3. Carniolian hybrid grafted with different genetic
origin larvae throughout the period extended from
4/7/2008 to 19/7/2008……………………..……….…………… 79
a. Acceptance percentage of grafted larvae………..…… 79

I
b. Amount of harvested royal jelly……………………...… 85
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly………...……….. 85
4. Carniolian hybrid grafted with different genetic
origin larvae throughout the period extended from
4/7/2008 to 19/7/2008…………………………………………... 89
a. Acceptance percentage of grafted larvae…………..… 89
b. Amount of harvested royal jelly……………………...… 94
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly…………...…….. 97
5. Egyptian hybrid grafted with different genetic
origin larvae throughout the period extended from
22/3/2009 to 6/4/2009……………………................................... 100
a. Acceptance percentage of grafted larvae……..……… 100
b. Amount of harvested royal jelly……………………...… 107
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly…...…………….. 109
b. Related grafted larvae………………………………...…….. 114
1. Egyptian hybrid used as donor and production
colonies throughout the period extended from
8/4/2009 to 20/4/2009……………………………………..……. 114
a. Acceptance percentage of grafted larvae…………… 114
b. Amount of harvested royal jelly………………………... 120
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly……...………….. 122
2. Italian hybrid used as donor and production
colonies throughout the period extended from
15/7/2009 to 2/8/2009………………………………...………… 125
a. Acceptance percentage of grafted larvae………..…… 125
b. Amount of harvested royal jelly……………………...… 131
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly……..………….. 134

II
3. Carniolian hybrid used as donor and production
colonies throughout the period extended from
15/7/2009 to 2/8/2009……………………………………..……. 137
a. Acceptance percentage of grafted larvae………..…… 137
b. Amount of harvested royal jelly…………………...…… 143
c. Effect of laying workers and number of combs
covered with bees on both accepted grafted larvae
and amounts of harvested royal jelly………...……….. 146
2. Determination of amino and fatty acids in some
samples of the produced royal jelly……………………. 154
a. Determination of amino acids………………………...… 155
1. Egyptian race used as production and donor
colonies………………...………………………………………….. 155
a. Essential amino acids……………………………..……… 155
b. Non essential amino acids……………………………… 157
2. Italian hybrid used as production and donor
colonies………………………………...………………………….. 159
a. Essential amino acids……………………………..……… 159
b. Non essential amino acids……………………………… 160
3. Carniolian hybrid used as production and donor
colonies……………………………………………………………. 162
a. Essential amino acids…………………………..…..……… 162
b. Non essential amino acids………………………...……… 163
b. Determination of fatty acids…………………………..… 171
1. Egyptian race used as production and donor
171
colonies…………………………………………………………….
2. Italian hybrid used as production and donor
colonies……………………………………………………………… 173
3. Carniolian hybrid used as production and donor
colonies……………………………………………………………… 175
SUMMARY…………….………………………………………………... 181
REFERENCES …………….……………………….……………….… 190
ARABIC SUMMARY …………….………………………………..

III
INTRODUCTION
Honeybee colonies have been used in the advantage of humans
as it used in royal jelly, honey, wax, bee venom and propolis
production, besides being very important as pollinator agents (Laidlaw,
1992).
Royal jelly (RJ) is one of the most valued products of honeybee
colonies and is produced by 6-12 days old honeybee workers (Apis
mellifera L), called nurse bees (Deseyn and Billen, 2005 and Hassan
and Khater, 2006)
It is secreted from the hypopharyngeal (watery) and mandibular
glands (milkey). Such glands are located in the workers heads (Haydak,
1970) and direct the development of honeybee larvae into queen bees
(Okamoto et aI., 2003) where it is an essential food for both.
honeybee workers do not stock RJ, being immediately utilized in
larval feeding. RJ has many uses such as: feeding workers and drones
larvae during a certain phase (Haydak, 1970); feeding the queen during
all its larval phase and adult life (Wang, 1965 and Wang and Moeller,
1969) and the cast differentiation between queen and worker bees is
related strictly to differences in the feeding of the larvae.
RJ is a white creamy substance with a slightly pungent odor and
taste. RJ consists naturally of a mixture of many constituents including
proteins (not less than 12%), carbohydrates (not less than 12%), lipids
(not less than 5.5%), moisture (not more than 66%) and 10-Hydroxy-
2-decenoic acid (10 HAD) (not less than 2%), (Howe et al.,1985).
Humans have used RJ for a long time for it's benefits as it's
believed that RJ stimulate the immune system, strengthens the body
and it's a good assistant cure for many diseases such as leukemia,
cancer, high blood pressure, high cholesterol, and infertility in males
and females (Krell, 1996).
RJ is being produced as a result of grafting process, and the
acceptance of grafted queen cups is being affected by type of nutrition
and queen cups introduced to the bees (Zeedan, 2002). Moreover,
acceptance percentage of queen cells was significantly higher when the
grafted larvae were less than 48 hrs old (Abd Al-Fattah et al., 2003 b)..
Mouro and Toledo (2004) found that Carniolan hybrid showed a higher
production of RJ per colony, in three-day collect when compared to the
Africanized honeybees and the Carniolan hybrid also had a higher
percentage of larvae acceptance compared to the Africanized
honeybees.
Furthermore, the total quantity of RJ produced by the colony
was higher in queenless colonies than in queenright ones (Ibrahim,
2002). Not only that , but RJ production is also being affected by bee
race (Saleh, 1999).
RJ is being commonly produced from the apiaries in Egypt for
its low costs and high price, since the price of one Kg could reach 6000
E.P. Natural RJ could be either stored under refrigeration for a period
extending from 18-24 months at – 20 ºC after packing or stored at 0 – 5
ºC and to save it's efficacy for one year.
As results of the increasing interest of RJ in human health, the
numbers of reports in the area of authentication and quality control
have increased within the past few years. Methods have been
developed to characterize the quality of RJ by determination of general

2
parameters as the main way to definite the quality of RJ is the chemical
analysis of its content specially the percentages of amino acids and 10
HDA.
Different factors may interfere in RJ production: genetics;
colonies internal population conditions; food flow and external
environment related to weather conditions (Nogueira-Couto, 1992 and
Azevedo-Benitez et al., 1998). Thus, it is important to study the
influence of these factors on RJ production, and honeybee’s regional
adaptability. The aim of this study was to investigate some factors
affecting the quantity and quality of produced RJ at Giza region as
follow:
1. Evaluation of some factors affecting royal jelly production in
honeybee colonies.
2. Determination of amino and fatty acid percentages in samples of
the produced royal jelly.

3
REVIEW OF LITERATURES

1. Methods of production
As Root (1945) in USA, reported in his book that in order to rear
good queens, it is necessary to have number of young -one day old-
larvae. Some authorities say that larvae at any time under three days
old from the egg will make good queens.
Snelgrove (1946) in England, used pure bees-wax in his
artificial queen-cups, but this wax must be fresh and best light color.
He also used 24 hrs. larvae for rearing queens.
Muller (1952) in Scandinavian region, grafted larvae within 24
hours of emergence.
Chauvin and Vuillaume (1955) in Paris, thought that the
acceptance of artificial queen cells was poor when the wax used in
making them contained queen extracts, which apparently inhibited
feeding of the grafted larvae.
Wilson (1955) in USA, placed about forty cells grafted with
twelve to fifteen hours old in swarm box. After twenty four hours, the
cells were transferred to a nursery colony. When the larvae were
between 3-4 days old, the greatest volume of royal jelly was obtained.
Vullaume (1956) reported that propolis mixed in the wax was an
inhibitor for queen cell acceptance.
Nienan (1958) proved that beeswax grafting compound yielded
better results than did asphalt emulsions
Eckert and Shaw (1960) in USA, mentioned that strong colonies

4
will start and finish 30 to 45 good queen cells.
Baryes (1963) gave a method that required the proper
preparation for a number of colonies. The first colony where queen
cups and eggs were produced had not more than 30-40 queen cells, of
which 10 - 12 were put into queen right colonies for rearing and finally
into queenless nuclei for emergence.
Inoue and Inoue (1963) in Japan, grafted larvae about 20 hrs-old
into the cups and concluded that the plastic cups and wax cups were
accepted (90.3, 75.8%, respectively). Plastic cups were superior except
in the yield of royal jelly per cup.
Abdel-Rahman (1964) in Egypt, indicated that capacity of the
colony to accept cell cups was averaged 18 cell cups in Alexandria and
34 in Gharbia. The highest records were occurred from March to
September, while the number accepted in February was clearly limited.
The same author (1967 b) found that artificial queen cups made
of new beeswax and of beeswax from old combs were equally
acceptable.
Ibrahim et al. (1969) in Egypt, reached a maximum of about 45
larvae at one time to the cups.
Mohanna (1969) in Egypt, found that the breeder colony has a
limited capacity to obtain numbers of successful queen cells differed
from one month to another. The highest percentage of successful queen
cells (averaged 71.8%) obtained in July. During this month, the total
number of introduced cells was 120 cups, and the accepted number of
queen cells was 86. The lowest average percentage of successful queen
cells occurred during March (45.8%). During May, the average

5
percentage of successful queen cells was 60%, as opposed to 57% only
in September. He mentioned that rearing queens from worker larvae
aged more than 48 hours proved to be less successful.
Lensky (1971) demonstrated that close contact of the queen with
the grafted larvae had no inhibitory effect upon their acceptance and
early rearing during late summer and winter. The acceptance
percentage of grafted larvae by colonies with free and confined queens
were 48-69 and 37-73%, respectively.
Wilson (1971) placed about forty cells grafted with 12 to 15 hrs.
old in swarm box. After 24 hrs., the cells were transferred to a nursery
colony. When the larvae were between 3-4 days old, the greatest
volume of royal jelly was obtained.
Weiss (1974) in Germany, found that the percentage acceptance
of double transplanted cells was 89% whereas that of single-
transplanted cells was 72%, where the bees were given a choice
between equal numbers of both presentations. He did not find this
difference to be a significant one.
Rekos (1976) in Italy, achieved best results for queen rearing
when larvae were transferred into prepared cups made from wax.
Moneim and Seleem (1978) in Egypt, found that the percentage
of successful queen cells in preliminary grafting with one day old
larvae.
Ebadi and Gary (1980) in USA, indicated some factors
affecting the acceptance of grafted queen cells. Acceptance was low
(32%) when queen pheromone (9-oxodec-trans-2-enoic acid) was
present in the wax of artificial queen cups. Statistically, significant

6
differences were found between the acceptance rate of queen cups
made of: (a) beeswax from old brood combs (86.6%), (b) fresh
beeswax (76.6%), (c) capping wax, beeswax foundation or equal parts
of paraffin and old beeswax (70%). Pure paraffin cells were not
accepted at all. The preference for queen cells in the upper part of the
nest is a surprising contrast with the normal position of queen cells in
natural nests, which most commonly are found on the bottom edges of
the comb.
Shawer (1980) in Egypt, showed that the highest percentage of
accepted larvae (90.00%) was obtained in queenless colonies and the
lowest (31.66%) was recorded when larvae were reared in queenright
colonies. Restricted queen gave an intermediate percentage (75. 00%).
Grandperrin (1981) found that the racial origin of larvae do not
affect transplant acceptance.
Li (1981) in China, found that worker honeybees prefer to feed
larvae in old cell cups. He recommended that new cups should not be
placed on the same frame, or put into the same colony, as old cups.
Larvae less than 24 hrs old should be grafted into cell cups which have
been used, and older larvae grafted into new cell cups.
Manino and Marletto (1981) in Italy, compared between wax
and plastic queen-cell cups for queen rearing and royal jelly
production. They found that the 2 types of queen cell cups were tested
by grafting in two queen starter-finisher co-operative colonies. No
statistically significant differences were found in the average
production per cell, or the total production, of royal jelly.
Zhang (1981) in China, stated that the amount of royal jelly

7
provided by nurse bees varied with the age of queen larvae. It was
small at 0-72 hrs of age, then it increased rapidly up to 96 hrs of age,
when 250 mg had accumulated, then it decreased gradually. He
recommended that 48 hrs old larvae should grafted, and the yield of
royal jelly harvested 36-48 hrs after grafting. This shortens the
conventional time required for a yield of royal jelly by one-third,
increased frequency of cropping, and eventually gave higher
production.
Zhu (1981) in China used the strong old combs with the middle
and lower portions removed as queen rearing frames. He measured the
ratio between workers and queen cells which were 1 comb of bees to 2-
3 queen cells. He said that a 10 -comb colony, with an average of 1000-
1500 nurse bees to feed each queen larva, could raise 20-30 queen cells
in one batch.
Shawer (1982) reported that the diameter of wax cups had no
effect on the acceptance of transferred larvae. He added that wax cups
added to rearing colonies 24 hours before grafting were easily accepted
more than those were not inserted to the rearing colonies, and the
highest average amount of royal jelly for one cell was found by rearing
50 cups in one colony as the amounts decreased by increasing the
number of the reared cells to 100-150 and 200 cells per colony. All
amounts of royal jelly produced from each treatment were increased by
increasing the number of wax cups.
Kither and Pickard (1983) concluded that some factors
promoting the acceptability of the artificial queen cups were being
continuously acquired during the first 26 h of the drawing process.

8
They reported that 24 hr. exposure of empty artificial queen’s cups to a
queenless colony before using them for grafting were significantly
improved their acceptability.
Fell and Morse (1984) studied the swarming in colonies in
single hive-body. They found a significant preponderance of cells in
the lower regions of the brood nest. The natural position may be chosen
not because of any inherent superiority but because its location where
brood is denser. They added that the queen cups can be built on comb
edges, and comb-building is active when bees are preparing to swarm.
Gracy (1985) in USA, reported a successful method for queen
rearing procedures involving grafting larvae that were put into the
starter colony when 24 hrs old and were then fed royal jelly by nurse
bees for 4 days.
Macicka (1985) in Czechoslovakia, carried out a series of tests
using 1-day-old larvae that were grafted into wax queen cell cups, with
or without royal jelly and put into a rearing colony. He noticed that the
percentage of larvae accepted varied between the top, middle and
bottom bars of the grafting frame, but there was no consistent pattern
from year to year. The development period was about 16 days in each
position. When the cups were placed 16 mm apart, acceptance was 80.3
% but at 30 mm apart only 70.1 %. Mean acceptance of larvae grafted
with royal jelly was 75.6% compared with 41.3% of those without
royal jelly (highly significant difference).
Cooke (1986) reported that the starter colony must be made on
the day of receiving the grafted cell cups. This is particularly important

9
if working with Apis mellifera capensis, as pseudo-queens may develop
during any delay.
Kirk-Visscher (1986) studied the kinship discrimination using
larvae and eggs for queen rearing. In first experiment, he transferred
larvae into royal jelly in queen cups, and had relatively high acceptance
(57%) and no evidence of nepotism. The bees reared nearly equal
number of unrelated larvae (77) and related nestmate larvae (76). In
second experiment, where eggs rather than larvae were transferred, the
acceptance was much lower (13%) and there was significant preference
for nestmate over foreign-hive eggs
Shawer et al. (1986) reported that the diameter of wax cups had
no effect on the acceptance of transferred larvae. They recorded that
wax cups added to rearing colonies 24 hours before grafting were
easily accepted more than those were not inserted to the rearing
colonies.
Weiss (1986) in Germany, stated that queen cups with grafted
larvae of Apis mellifera carnica, were introduced a starter colony and
removed after 3-4 days for studying the royal jelly and larvae. With
subsequent batches, the percentage acceptance decreased as did mean
weights of larvae reared and of royal jelly extracted from the cell.
Skowronek and Skubida (1988) in Poland, found that grafting
was very successful in cups containing fresh royal jelly or royal jelly
plus water (1: 1), but was poor in cups containing honey or water or
nothing. On introduction to queen rearing colonies, queen larvae were
accepted best (77.5 %) by colonies without brood; acceptance was 57.5
% in colonies with sealed brood and only 42 % in colonies with both

10
sealed and unsealed brood. Queen larvae were fed significantly more in
colonies without unsealed brood, and maximal queen weight was high
(160 mg).
In Egypt, Sharaf El-Din and EI-Samni (1990) studied the effect
of grafted queen larvae in queen cell cups made of rubber or bees wax
on queen rearing. They found that the acceptance percentage of grafted
larvae and total royal jelly production per colony were significantly
higher in colonies with rubber cups; the amount of royal jelly per cup
was also slightly but not significantly, higher in the rubber cups.
Chang and Hsieh (1991) in China, recorded that royal jelly
production is influenced by several factors, of which the major ones are
colony size and number of royal jelly collection bars set in the colony.
Lawrence and Cobey (1991) In USA, reported that the number
of queen cells given to a cell builder colony was important. Queen
producers routinely grafted 30 to 80 cells per cell builder. However,
fewer queen cells given to each cell builder increased the likelihood of
the cells receiving a more plentiful diet. Depending on the specific
queen rearing system used, cell builders giving more than 45 cells may
produce inferior queens.
Moreover, Chen and Chang (1993) determined the effect of
different numbers of queen cell cups on royal jelly production at 2 and
3 days after grafting. Queenright colonies of a selected A. mellifera
stock (20000 bees/7 -frame colony) were given 64, 96, 128, 160 or 192
golden-yellow plastic cups in 2-day royal jelly production trials in
March, or 68, 102, 136, 170 or 204 cups in 3-day trials in July. Mean
weight of royal jelly/cup was decreased with increasing number of cups

11
in both trials. The total production was higher at 3-day post-graft than
at 2-day (553.1±79.1 mg vs. 399.5±52.1 mg).
Abd Al-Fattah (1994) in Egypt, recorded that the tendency of
queenless colonies toward queen rearing by constructing queen cells
was gradually decreased with increasing the number of introduced
queen cells to each colony.
Ali (1994) in Egypt, studied the type of cell bar and its level on
the frame in the colony. Four experiments were conducted: wide cell
bar holding 45 queen cups and fixed on the upper position (A); wide
cell bar holding 45 queen cups and fixed on the second position from
the top (B); three normal cell bars each contained 15 queen cups were
fixed on the third following position from the top (C); and three normal
cell bars each contained 15 queen cups were fixed on the third
following position from the bottom (D). The highest numbers of
accepted queen cells were found in the D, B and C positions, while the
highest amounts of royal jelly were obtained in the C and D positions.
In addition, he concluded that grafting with 12 or 36 hrs old larvae had
greatest volume of royal jelly than obtained after 2.5 days. The best
number of cell cups was 45 cups per a colony. He obtained the
maximum yield of royal jelly from strong colonies contained ten combs
covered with bees, and followed by those contained seven covered
combs. He reported that the amount of royal jelly collected from one
queen cell was larger in strong colonies than those of weak ones.
In Albania, Dedej (1994) compared between double-grafted and
single one on acceptance of grafted larvae and number of emerged
queen. He found that all these parameters were higher in the double-

12
grafted than in the single one.
El-Sherif et al. (1994) in Egypt, detected that the highest
amounts of produced royal jelly taken from four successive batches
were 29.01 and 28.80 g/colony when 36 or 12 hrs old larvae were
grafted and the royal jelly collected after 2.5 days, followed by 24.53
g/colony when 60 hrs old larvae were grafted and the experiment
durated 1.5 days. The lowest amount was collected after 3.5 days.
Toor and Littlejohn (1994) in New Zealand, reported that
harvested royal jelly 66 hrs after grafting larvae resulted in similar
yields to that harvested at 72 hrs or 78 hrs, indicating that beekeepers
can harvest at any time on the third day without affecting production.
The chemical components of royal jelly within certain quality standards
were not affected by the treatments. Also, they noticed that mean yield
of royal jelly per harvest and cell acceptance were much lower for the
double (2nd) harvest than for the double (1st) harvest and these in turn
were significantly lower than for the single harvest.
Abd Al-Fattah (1996) in Egypt, studied some of the factors
affecting the acceptance of grafted queen cell cups and mass production
of royal jelly (RJ) in honeybee nursing colonies. Acceptance
percentage and mean amount of RJ were not affected by the presence
of restricted young or old queens under the compartment of queen
rearing. Highest rate of acceptance was attained using 48 hrs-old
grafted larvae followed by 24 hrs larvae. Similar quantities of RJ were
obtained after nursing the older larvae for 2 days and younger ones for
3 days. The lowest amount was collected after nursing 24 hrs-old larvae
for 2 days. The nurse honeybee win nursing colonies which prepared

13
by displacement or shaking gave the best results for the acceptance of
grafted larvae and mean quantity or RJ produced per queen cell cup or
colony. Empty queen cups that partially drawn by placing them in a
colony 24 hrs before using in grafting were significantly increased the
acceptance of grafted larvae and RJ production.
In India, Rana et al. (1996) found that the yield of royal jelly
was related to the weight of royal jelly per cell and the number of
queen cups in a colony.
Abo El-Kassem (1997) in Egypt, observed that the highest
amount of royal jelly was recorded from queenless treatment (2.25g /
colony), followed by that of caged queen (1.4), while requeening came
the third (1.05) and reduce combs gave the least amount of royal jelly
(0.02g / colony). However in both treatments, increase combs and
excluder, led to no royal jelly was collected at all.
Laidlaw and Page (1997) in USA, found that approximately 15
minutes to one hour period is required for the first graft of 3 or 4 bars
of cells (45 to 64 grafted cell cups) to the cell builder. Two frames with
grafted queen cell cups (90 to 128 cells) may be given at one time.
They are removed 24 hours later and distributed to the finishing
colonies.
Chuda-Mickiewiez and Prabucki (1998) found that the
effectiveness of queen rearing examined in 1996 and 1997 was more
favorable when queens were reared from 1-day-old larvae (56.03%),
than from eggs (39.99%).
Tarpy and Fletcher (1998) demonstrated that acceptance of wet-
grafted 1-day- old larvae was independent of their position in the brood

14
nest. Result of pre-emergent aspect indicated that nurse bees do not
functionally discriminate between related and unrelated larvae during
queen rearing.
In Turkey, Durmus and Guler (1999) determined the effects of
different colony population sizes, queen cup types and harvest times on
royal jelly production in A. mellifera colonies. They found that the
average amounts of royal jelly were 369.90& 402.40 mg/cup at 10& 20
frames; 401.26& 371.04 mg/cup with pure and commercial wax cups
and 343.35& 428.95 mg/cup at 48 &72 hours post larval transfer,
respectively. It appeared that colony size, cup type and harvesting
interval had significant effects on royal jelly yield (P<0.01).
Saleh (1999) in Egypt, used Doolittle and Jenter methods for
producing Royal jelly and queen cell cups. He concluded that in
general the Jenter method was better than Doolittle one under all cases.
In China, Jianke (2000) noted that the royal jelly production can
be done only if colony's population is big even when few honey plants
were in bloom.
Li (2000) used five wood bars on each royal jelly frame for a
total of 125-170 queen cells, and 69 - 100 gm of royal jelly can be
collected from each frame one cycle that lasting 72 hours.
In Egypt, Sharaf El-Din et al. (2000) detected that installing
grafted queen cups on the lower level of bars induced the highest
percentages of acceptance of grafted larvae. The least results were
obtained when grafted queen cups were fixed on the upper bar. The
differences between effects of the three levels (lower, middle & upper)
of bars were mostly significant.

15
Tarpy et al. (2000) found that the difference in larval acceptance
was highly significant between new hatching larvae and 2-day-old
ones.
Ismail (2001) in Egypt, noticed that the treated colonies with
volatile oils produced more total numbers of queen cells in comparison
with the control. Treated colonies with rosemary produced 178 queen
cups all over the year (94 cups during spring), while those treated with
camphor produced 144 queen cups (91 cups during spring), however
colonies treated with thymus produced only 74 queen cups in the same
season from the total annual number (136 queen cups).
Moreover, Ibrahim (2002) in Egypt, showed that the total
quantity of royal jelly produced by the colony was higher in queenless
colonies (7.54 gm/colony) than in queenright ones (5.67 gm/colony).
Kruk and Skowronek (2002) reared queens during the period
from 1990 to 2001 and they found that the percentage of capped queen
cells varied substantially over the years with the lowest rate of 32.9%
in 1990 to the highest rate of 58.1% in 1999.
Wilkinson and Brown (2002) reported that the mean acceptance
rate of 6666 grafted larvae in queenright colonies from 1989 to 2001
was 81%.
Zeedan (2002) in Egypt, summarized that the acceptance
percentage of grafted larvae was higher in wax cups (78.3%) than in
plastic ones (77.2%).
Abd Al-Fattah et al. (2003a) reported that the acceptance
percentage of queen cell was significantly higher when the grafted
larvae were 24 hours old, followed by 36 hours old then by 12 hours

16
old. Also, they added that double grafting technique attained the
highest percentages of acceptance.
Emsen et al. (2003) observed that the larval accepted rates were
77% and 70% in the single and double grafting, respectively.
Dodologlu et al. (2004) reported that the average acceptance rate
in queenless colonies was 95%.
Cengiz et al. (2009) found that the average acceptance rate in
queenless colonies in June, July and August were 95, 86.6 and 78.3%
and in queenright colonies were 78.3, 71.6 and 61.6%, respectively.
2. Effect of season on royal jelly production
Maurizio (1950) stated that the physiological conditions of
summer or winter bees did not depend on the time of the year, but was
a reaction to the living conditions in the colony.
Under Scandinavian conditions, Muller (1952) showed that the
hot summer limited the queen rearing period to two months. Moreover,
it made impossible to produce large number of bee queen rearing.
Hanna (1963) in Egypt, studied the queen rearing in
experimental stocks which carried out in August, October, January and
April. He found that the different seasons had only an insignificant
effect on the acceptance of queen cells, although the percentage of
acceptance was somewhat higher in the spring and summer than in the
autumn and least in the winter. On the other hand, the amount of royal
jelly was significantly affected by the season. The largest amount was
obtained in the spring, followed by the summer and the lesser amount
was collected in the winter. Taking the amount obtained in winter to be
100, the relative figures for the autumn, summer and spring would be

17
194, 219 and 253, respectively.
Abdel Rahman (1964) in Egypt, indicated that the capacity of
the colony to accept cell cups was 18 cell cups on average at
Alexandria and 34 at Gharbia. The highest records occurred from
March to September while the number accepted in February was
clearly limited.
Fotti et al. (1971) stated that the acceptance of honeybee queen
cells was highest (about 88%) in August but lowest (about 45%) in
May and was insignificant by their position in the hive.
Ewies (1974) in Egypt, stated that the Carniolian hybrid bees
that used for royal jelly production gave 11.07 gm in early summer,
8.23 gm in late summer and 5.43 gm in the autumn.
In Arizona, USA, Taber (1979) found that in hot weather
queenless colonies did not build queen cells.
In Turin, Italy, Manino (1982) studied the effect of temperature
and rainfall on queen rearing, during May to September of 1980 and
1981. He reported that successful queen mating required good weather,
but grafting and the development of queen cells were only slightly
affected by climatic conditions.
Krol (1985) observed that the best time of the year for queen
production was at the end of May and in early August. The amount of
royal jelly in the cell was the maximum on the fourth day of
development.
Nour (1992) in Egypt, noticed that the production of queen cells
in the spring and early summer may represent an index to the tendency
of the colonies to prepare for swarming. He showed a positive

18
correlation with queen cells production in the spring and early summer.
The number of queen cups/col. indicated a negative correlation with
queen cells in spring, whereas positive correlation was recorded in the
summer.
In Saudia Arabia, El-Sarrag (1993) found that the highest
percentage of accepted queen cells (92%) were in February and during
March- June (82%). The least favorable periods (48%) were in August
-September and (28%) in December- January.
Ali (1994) in Egypt, concluded that Royal jelly production was
higher in the summer (mid June to early October) than the spring (end
March) or the autumn (late October) and the winter (December).
In UK, Gue (1998) suggested that as the nurse bee population
reached its peak in mid summer in healthy, mature and populous
colonies, the pheromone level becomes an impelling force leading to an
instinctive urge to rear queens and swarm.
In Yemen, Al-Humyari et al. (1999) concluded that about 69.07
and 96.91% of annual counts of periodical newly constructed queen
cells (23.12 and 56.34 cells/colony) were in the winter nectar flow in
Yemeni and F1 Carniolian colonies, respectively.
Ahmed (2000) in Egypt, mentioned that under the
environmental conditions of Al- Kome El-Akhdar, Benha region, the
highest number of accepted queen cells was obtained during the cotton
and clover nectar flow seasons for Carniolan and Italian races,
respectively.
Sharaf El-Din et al. (2000) revealed that the production and
quality of queens were affected by the rearing season and they recorded

19
an increase in larval acceptance during the spring than summer season.
Taha (2000) observed that, under the environmental conditions
of Kafr El-Sheikh region, the highest level of building queen cups,
queen cells, and the highest amount of royal jelly collected throughout
the blooming seasons of citrus, clover and cotton was observed in
colonies moved to food sources-rich areas. The highest amount of royal
jelly was collected during April (the blooming season of citrus),
followed by May (clover season).
Zohairy (2001) proved that under the environmental conditions
of El-Mansoura region, the mean percentages of accepted queen cells
in a descending order were in May, June, July, April, September,
August and March in 1998. In 1999 they were in June, July, August,
May, September, April and March.
Under the environmental conditions of Giza region, El-Waseef
(2002) showed that the highest level of building queen cups was during
March. At the same time the highest average amount of royal jelly (6.2
gm/colony) was collected in April from a colony contained 544 inch2
of worker sealed brood. While the lowest amount (4.5 gm/colony) was
collected in August from a colony contained 490 inch2 of worker sealed
brood.
Zeedan (2002) in Giza region, Egypt found that the highest level
of building queen cups was during March (24.6 queen cups/colony),
while the lowest level was during December (0.9 queen cups/colony).
The building queen cups was higher during the spring followed by
summer, winter and autumn seasons, whereas the percentages of queen
cups were 38.4, 26.4, 20.1 and 15.1 % during the aforementioned

20
seasons, respectively. He added that there were significant differences
in the mean of accepted larvae between both the spring (84.2%) and
summer (82.3%) from one side and both the autumn (73.4%) and
winter (71.1%) from the other one.
Abd Al-Fattah et al. (2003b) in North Sinai, Egypt determined
that the highest rate of acceptance was occurred during the summer,
represented 82.8%. Significant differences were found between the
summer and other seasons. The lowest significant percentage was
appeared in the winter (36.1 %). Moderate results for the accepted
larvae were noticed during the spring (70.0%) and autumn (72.8%)
seasons with non-significant differences between them.
Also, Serag EI-Dein (2004) in Kafr El-Sheikh, Egypt mentioned
that the highest amount of royal jelly was produced during the period
from 15th May until 15th July (clover nectar flow), followed by cotton
nectar flow period (1st July to 30th August) while broad bean nectar
flow period (15th February until 15th April) was the least one.
Genc et al. (2005) observed that the maximum acceptance was
in dry grafting (98.3%) in July and grafting with addition royal jelly
was (93.3 and 91.7%) in July and August, respectively.
Guler and Alpay (2005) found that the mean acceptance rates of
carnica and ligustica grafted larvae from May to August were 76.11
and 78.89%, respectively.
Sahinler and Kaftanoglu (2005) in Turkey, studied the effects of
season on the acceptance rates of the larvae and royal jelly production.
The average acceptance rate and royal jelly production per cell
throughout the season were 88.2% and 0.263g in queenless cell

21
builders and 72.1% and 0.214g in queenright cell builders, respectively.
Taha (2005) showed that the mean percentage of accepted queen
cells was affected by nectar and pollen sources during the blooming
periods of faba bean , citrus , squash , clover and cotton in the different
sites during the years of 2002 and 2003. Also, he concluded that the
highest percentage of accepted queen cells (55.00 & 55.5%) was
noticed at the clover blooming period then 53.33 & 54.45% during the
citrus flowering period in 2002 and 2003, respectively.
Under environmental conditions of Giza region, Hammad
(2007) recorded that production of queen cells started in the 25th of
March 2006 and increased gradually till reached the maximum during
the period from the 20th of May to the 17th of June, 2006 in the tested
colonies, then the mean number gradually declined until the 9th of
November, 2006 where no queen cells were present. The mean number
of queen cells produced/colony during the spring season was higher
than in the summer one.
Shawer et al. (2007) in Kafr El-Sheikh and El-Behiera
Governorates, reported that the highest percentage of accepted queen
cells was obtained from colonies located in Rasheed region during the
citrus flowering period, followed by that located in Ammya region
throughout the clover blossoming period. On the other hand, the lowest
percentage of accepted queen was recorded during the squash blooming
period in Kafr El-Sheikh region in the two years, respectively.

22
3. Effect of genetic origin on royal jelly production
Laidlaw and Eckert (1950) noticed that the Italian and
Caucasian races tend to produce fewer queen cells than other races
such as Carniolian.
Giordani (1958) concluded that the yeast has about the same
nutritive value as pollen for feeding bees, judged by hypopharyngeal
gland development.
Pickard and Kither (1983) found that the acceptance percentage
of black larvae (with characteristics of A. m. mellifera) and yellow
larvae (with characteristics of southern European races) reared in
starter colonies intermediate between black and yellow races were 81%
and 74%, respectively.
Breed et al. (1984) found no significant differences in the
number of larvae accepted or number of queen reared from A. m.
mellifera or A.m. carnica. They concluded that the workers were
incapable of discrimination between larvae on the basis of relatedness.
Page and Erickson (1984) found that when workers rearing
queens were given a choice between which were their full sister, and
larvae from another colony to which they were related approximately
as closely as they would be to their half sisters, they preferred to rear
the more closely related larvae. This discrimination may have been
facilitated by differences in colony odor between the two groups of
larvae.
KroI (1985) in Poland, found that during the first 6 days of
queen larval development, differences were observed between
Caucasian, Carniolian, Italian and North European honey bee races.

23
Carniolians built the largest and heaviest queen cells and put most royal
jelly in each cell. In all the races, the amount of royal jelly in the cell
was maximal on the fourth day of larval development.
Under the local conditions of Alexandria region, Yakoub (1998)
stated that both the Carniolian and Egyptian bees produced large
numbers of queen cells during the year, while the Italian race built only
one cell cup during July
Saleh (1999) in Egypt, showed that the 1st hybrid of German bee
race was the best tested one for collected royal jelly, followed by the
Carniolian bees, while the third one was the Italian bee race.
Mohammedi and Le Conte (2000) showed that A. m. mellifera
workers accepted significantly more sisters (77.5%) than alien bees
(20%). Also, A. m. ligustica workers accepted 77.5% of sister workers
and only 45% of mellifera workers. This difference was significant
confirming the ability of the bees of the two races to discriminate.
Mouro and Toledo (2004) in Brazil, detected that the hybrid
Carniolan showed a higher production (P=0.0001) of royal jelly per
colony in three-day collect (3.95 ± 2.92 g) when compared to the
Africanized honeybees (2.23 ± 2.3 g). The Carniolan also had a higher
percentage of larvae acceptance (55.45 ± 36. 7%) compared to the
Africanized honeybees (35.8 ± 28.3%).
Tarpy et al. (2004) reported that when colonies rear queens,
there may exist a small amount of conflict over which individuals to
raise as queens based on genetic relatedness, but the workers mostly
cooperate in constructing queen cells so that queens which emerge
were of high reproductive quality.

24
Sahinler and Kaftanglu (2005) in Turkey, proved that Mugla
bees was found to be more suitable for royal jelly production than the
Caucasian bees under the experimental conditions.
Albarracin et al. (2006) used two genetic groups Africanized
and Italian bees and three treatments of grafting; T1 (larvae from the
same genetic group and the same colony), T2 (same genetic group and
different colonies), and T3 (different genetic groups). They found that
in the process of queen production, the genetic groups and treatments
showed no significant differences in acceptance percentage of larvae
and amount of royal jelly for queen cells.
4. Effect of feeding on the produced royal jelly
Dietz (1969) found that, the development of glands such as the
hypopharyngeal glands, depends upon adequate amounts of protein in
the honey bee’s diet. During the early adult life of worker bees, all
nitrogen was derived from pollen protein. Consequently, young bees
must consume a large quantity of pollen in the first two weeks of their
adult life. Some worker bees began consuming pollen within one to two
hours after emergence. Within twelve hours after emergence, 50% or
more of the workers have already consumed pollen in small amounts.
Knox et al. (1971) in USA, fed newly emerged adult bees with
pollen incorporated as 5% by weight in honey-sugar candy (2.5). Each
cage of bees was provided with water and kept at 30° C and about 65%
relative humidity. They indicated that the average span life of bees fed
dandelion pollen was about 32 days (about the same as the control
bees) but their hypopharyngeal glands failed to develop. Bees fed either
blueberry or cranberry pollen lived about two weeks longer and their

25
hypopharyngea1 glands developed well.
Peng and Jay (1976) in Canada, used groups of 400, five days
old nurse bees to rear queens in cages in the laboratory. Seven different
diets were fed to the nurse bees. The development of the
hypopharyngeal glands of the nurse bees, their queen- rearing ability
and the brood production of queen that they reared were used to
evaluate these diets. Diets of stored pollen fortified with amino acids or
vitamins, and of fresh pollen had similar effects on the development of
the hypopharyngeal glands. The hypopharyngeal glands of nurse bees
fed pollen degenerated with age, queen rearing by these groups was
low.
Szymas and Torgowski (1978) in Poland, tested the extracts of
groundnut oil meal, soybean, and mashes from oil seed rape flax and
lapin as pollen substitutes. The amount of each substance mixed with
sugar candy was such that the mixture contained 10% crude protein.
The value of each feed was assessed by the development of the
hypopharyngeal glands of young workers. The only feed which gave
reasonable results was that contained groundnut oil meal, but the gland
development was less than that in the control groups fed on candy
containing bee- stored pollen.
Atallah (1983) in Egypt, tested high protein food stuffs,
including torutein (torula Yeast), alfalfa protein extract (type 1), alfalfa
protein extract (type II), chickpea meal, and broad bean meal against
almond pollen as the control. Newly emerged worker bees were kept
for two weeks in a controlled laboratory condition and were fed with
diets contained 10% of each protein. The food consumption, the

26
developmental index of bees, hypopharyngeal glands, the relative
protein utilization, and the relative nitrogen excretion were used as
screening parameters. He found that worker bees fed with torutein and
alfalfa protein extract (type I) had significantly higher food
consumption, gland development index, gland protein content and
relative protein utilization. he suggested that (tortutein) and (type I) of
alfalfa protein extract were better protein supplement for feeding bees.
Shuel and Dixon (1986) found out an artificial diet which
stimulates royal jelly and can be used for the rearing of honey bee
larvae. The diet is chemically defined except for protein from royal
jelly; it also contains 10- hydroxyl-2-decenoic acid (10HDA) from
royal jelly. Larvae died early when fed on diets containing egg albumen
or enzyme-hydrolyzed casein, or when 5-6% of the protein complement
was replaced by free amino acids. Both an inorganic and organic
phosphate source was necessary. Development was normal when half
of the 10-HDA had been replaced with 9-oxo-2-decenoic acid.
D'l Albore et al. (1987) In Italy, reared groups of newly emerged
workers on test diets (in the absence of brood and the queen). A sample
of bees was taken from each group at regular intervals from three day
to three weeks, so that hypopharyngeal gland development could be
measured. The largest development was detected in bees fed on either a
spring pollen mixture or Papaver robur or (equally effective) Juglans
regil1i Cisturhoeas pollen. The other pollens tested for gland
development were Quercus spp and typha latifolia. Development was
poor in bees fed on maize gluten meal, and bees died within six days
when fed on alfalfa leaf meal. Hypopharyngeal gland size was greatest

27
in 10-20 days old bees. They also found that the right gland grew
significantly more than the left one. The maximum development of
gland was reported between 18-27 days.
Macicka (1987) in laboratory experiments stated that honeybees
fed with carbohydrate containing protein additives had larger acini in
the lobules of the hypopharyngeal glands, than bees fed on diets
without protein additives of 23 tested feeds. The most effective were
sugar candies with the addition of dried yeast, dried defatted milk, fresh
pollen pellets and bee bread.
Nour (1988) in Egypt, concluded that the enzymes activity
(gluco-oxidase, invertase and diastase) of the hypopharyngeal glands of
worker bees were markedly differed at different ages during different
times of the year; they were higher in old bees more than 14 days old.
While they were lower in young bees (lees than 14 days old). In
addition, the enzymes values in the hypopharyngeal glands of the
worker bees depended on the food consumption which is rich in pollen
or pollen and supplements during the season flow. Food rich in pollen
help the redeveloping of the hypopharyngeal glands in worker bees
especially in old bees (foraging bees).
Mohamed and Omar (1992) in Egypt, documented that the
development of the hypopharyngeal glands is a parameter reflects the
nutritive quality in honeybee feed.
Chang et al. (1993) In Taiwan, stated that the Chinese formula
and natural tea pollen showed improved results in both food
consumption and production of royal jelly in the treatment with setting
a pollen trap for Chinese formula and with setting out the pollen trap

28
for natural tea pollen, respectively.
In Egypt, Ali (1994) recommended that honeybee colonies must
be provided during dearth or the set on of the spring with yeast alone or
mixed with sugar syrup, cakes of soybean flour fortified with pollen or
any pollen supplements to increase the percentage of accepted larvae.
Atallah et al. (1995) in Egypt, concluded that feeding queen
rearing colonies with sugar syrup and pollen supplement increased
royal jelly yield and its protein content.
El-Dakhakhni and Metwally (1995) in Egypt, reported that the
development of the hypopharyngeal gland was promoted by high
protein concentration of the diet.
Szymas and Przybyl (1995) fed six pollen substitutes with an
amino acid composition similar to that of pollen to groups of test bees
in the laboratory. They found that bees on all diets showed satisfactory
development of hypopharyngeal glands, and 2 diets gave results
comparable to those of pollen. Those were; (A) a mixture containing
powdered hen's eggs and with 10% protein content, (B) a mixture
containing many substance, including hydrolyzed casein, skimmed
milk, eggs, torula yeast and ground soya beans, and with 26% protein
content. Feeding colonies with pollen substitutes in the spring
promoted early and in the autumn bees fed on pollen substitutes had
larger hypopharyngeal glands.
Abd Al-Fattah (1996) in Egypt, stated that, about twice amount
of royal Jelly was yielded from colonies provided with pollen
supplement paste than those from unfed ones. The nurse honeybee in
nursing colonies which prepared by displacement or shaking gave the

29
best results for the acceptance of grafted larvae and mean quantity of
RJ produced per queen cell cup or colony.
In Egypt, El-Sherif et al. (1996) indicated that the highest
percentage of accepted queen cells was found in colonies fed on a 50%
sucrose syrup beside sucrose yeast cake (49.31 cells/graft/colony with
82.19% acceptance) or on honey syrup beside sucrose-pollen cake
(45.06 cells/graft/colony with 5.10% acceptance). The lowest
percentage was obtained in colonies offered fructose syrup beside
fructose-yeast cake (33.31 cells/graft/colony with 55.52% acceptance)
or offered stored honey and pollen in combs (30.69 cells/graft/colony
with 51.15% acceptance).
In Poland, Szymas and Przybyl (1996) used potato protein,
soybean meal, yeast, low-fat powdered milk, powdered chicken eggs,
extruded maize and vitamin mix to feed honeybee colonies. They found
that the development of hypopharyngeal glands was be similar to those
of the bees that were fed bee bread.
El-Shemy (1997) in Egypt, concluded that building the queen
cups was higher in fed colonies than unfed ones (the control).
In Egypt, Sharaf El-Din et al. (1999) fed queen rearing colonies
on yeast, soybean, mandrin cortex jam, semi-dry date and sugar syrup.
They detected that the tested foods induced highly significant increase
in the acceptance percentages of grafted cups, sealing queen cells. The
tested materials could be arranged descendingly according to their
efficacy as follows; yeast, soybean, mandrin cortex jam, semi-dry date
and sugar syrup.
In Egypt, Ahmed (2000) summarized that the number of

30
accepted queen cells was higher in case of brewer's yeast and sugar for
both the Carniolian and Italian races.
Gencer et al. (2000) in Pakistan, used three groups of
supplemented feeding in queen rearing; (A) one liter syrup (sugar:
water, 1:1), (B) one liter syrup with 3 g vitamin mixture, (C) one liter
syrup with 10 g fresh pollen and (D) the control group. They found that
the acceptance percentage of larvae grafted were 72.9, 85.4, 83.3 and
69.8% in the group A, B, C and D, respectively and significant
difference was found in the acceptance% of grafted larvae. The
acceptance % of grafted 1-day and 2-day old larvae were 73.4% and
82.3%, respectively. They also mentioned that the acceptance rate was
not affected by larval age and the difference between them was not
statistically significant.
Pernal and Currie (2000) suggested that development of
hypopharyngeal glands was strongly correlated with the amount of
protein workers consumed from pollen diets. Also, they proved that the
development of hypopharyngeal glands varied with diet and
collectively proved to be sensitive measures of protein utilization and
pollen quality.
In Egypt, El-Waseef (2002) indicated that the fed colonies were
active in building more queen cups than unfed ones.
Mansour (2002) on his work related to the development of
hypopharyngeal glands, found that the interaction between diet C ( 200
gm. dried brewer's yeast + 200 gm. skim milk powder + 400 gm.
soybean flour + 350 gm. clover extract + 1000 gm. sugar powder) and
12 days old workers was the best on the development of the

31
hypopharyngeal glands during the first year. During the second year,
the interaction among diet C and 18 days old was the best in the spring
season. In the autumn, the interaction among diet C and 12 and 18 days
old and the interaction between diet A (200 gm. dried brewer's yeast +
300 gm. com flour + 400 gm. chick pea flour + 100 gm. bran + 1000
gm. sugar powder + 500 ml. water) and 12 days old and the interaction
between diet B (4 gm. dried brewer's yeast + 200 gm. agwa + 300 gm.
wheat bran + 10 gm com flour + 1000 gm. sugar powder + 500 ml.
water) and 18 days old, all were the best. In the winter, the interaction
between diet C and 11days old was the best.
Mohammed (2002) concluded that feeding rearing colonies on 2
parts bean flour; 1 part date paste: 1 part skimmed milk gave the best
results of acceptance during the spring, summer and autumn seasons, as
attained 88.21, 87.77 and 72.23 %, respectively. He also found that, the
maximum of 10% brewer's yeast: 3 chick pea flour: skimmed milk
induced the highest mean acinal surface area (0.03868 mm2). While the
lowest acinal surface area (0.01389 mm2) was detected in caged bees
fed on sugar syrup only.
Abd Al-Fattah et al. (2003b) reported that the highest
percentages of acceptance was obtained when the queen rearing
colonies were provided continuously with yeast cake fortified with
pollen or sucrose, while offering the stored honey combs only gave the
lowest results.
Taha (2005) studied the effect of nectar and pollen sources on
mean number of lobules/1 nun and mean acinal surface area (mm2) of
the right hypopharyngeal gland in workers collected on the middle cell

32
bar of queen rearing colonies during the blooming periods of faba bean,
citrus, squash, clover and cotton in the different sites during the years
of 2002 and 2003. He proved that the mean number of lobules/l mm
and mean acinal surface area were highly significant; higher in colonies
moved to rich nectar and pollen sources locations than non-moved ones
during all blooming periods in the two years.
Under Giza environmental conditions, Hammad (2007) found
that the mean acceptance percentage recorded higher values for
colonies fed with liquid yeast diet (Candida tropicalis) at 25%
concentration than in colonies fed with sugar syrup in both the spring
and summer seasons.
In Egypt, Shawer et al. (2007) reported that the percentage of
accepted queen cells was significantly higher in colonies moved to
sufficient nectar and pollen locations than non-moved ones during the
all flowering periods in both years.
Hammad et al. (2009) in Egypt, stated that significant
differences in royal jelly production between naturally queenless
colonies fed with liquid yeast diet (Candida tropicalis) at the
concentration of 25% and colonies fed with sugar syrup only were
detected during summer season. The mean number of queen cells/
colony was higher in the queenless colonies fed with yeast diet than
those fed with sugar syrup only in both spring and summer seasons.

5. Development of hypopharyngeal glands


Maurizio (1954) in USA, declared that the hypopharyngeal
gland and fat bodies development reached the maximum between 20 to
27 days and retrogresses in their development. If the physiological state

33
of bees is investigated too early, after 8 to 14 days, false conclusions
were easily drowned as to the effectiveness of the tested substances.
What (1954) in Germany, found that brewer's yeast was a
suitable pollen substitute in periods of dearth, as its composition and its
effect on the development of the pharyngeal gland.
Standifer et al. (1960) in USA, reported that the rate of
hypopharyngeal gland development seems to be related to protein
content of the diet.
Ricciardelli et al. (1987) found that the right hypopharyngeal
gland grew significantly more than the left one. The largest glands
development was observed in bees fed on a spring pollen mixture.
Development was poor in bees fed on maize gluten meal.
Takenaka (1988) reported that nurse worker bees 10 and 14 days
old synthesis had the highest rate of protein in their hypopharyngeal
glands. During this period, they were producing protein-rich royal jelly.
After the 14th day, protein synthesis decreased as does the synthesis of
royal jelly protein.
Yousif et al. (1988) in Egypt, observed that the hypopharyngeal
glands have two peaks of activity; the first took place between 6-9 days
and may extended to the 12 days, while the second took place at the 18
days.
Also, Mohamed and Omar (1992) in Egypt, reported that the
hypopharyngeal glands had two peaks of activity. The development of
hypopharyngeal glands a parameter reflects the nutritive quality in
honey food with best assimilation in gut was determined after feeding
bee workers on sugar syrup containing different formulations of

34
Bacillus thuringensis.
Gracioli et al. (1999) in Brazil, reported that the protein profiles
of the hypopharyngeal glands of aged A. mellifera workers (40 days
old) experimentally induced during the later phase of their adult life (to
return to the task of brood care due to the absence of younger bees
“experimental condition”) were presented. The pattern was similar to
that of 12-days old workers reared under normal and experimental
conditions, suggesting glandular reactivation. A different and specific
electrophoretical pattern was observed in 40 - days old workers from a
normal colony.
Malone et al. (2004) kept newly-emerged adult honey bees in
cages and fed it for ten days with a pollen-food to which one of the
following proteins had been added at concentrations equivalent to
pollen expressing 1.25% (of the total protein) aprotinin, 11.2 μM avidin
or 0.3% Cry1Ba protein. The control bees were fed pollen food without
additive, with casein added, or pollen-candy. None of the treatments
significantly affected bee survival, or the mean diameters of gland
acini, gland mass or protein content on days 1, 4, 8 or 10. Bees
consumed significantly more of the pollen-candy than the other foods.
Thus, effects of these transgene products on gland development are
unlikely, but field trials using transgenic plants are required for a full
assessment.
Deseyn and Billen (2005) showed that a secretory cycle within
the cells, although production of secretion is asynchronous between
different cells within an acinus. Secretory vesicles appeared in 3 day
old bees, while the peak production was around 6 days. Thereafter, the

35
volume of the acini as well as the number of secretory vesicles
decreased and no vesicles are visible after 3 weeks of age. Foragers
display degenerative structures in their cells. The hypopharyngeal
gland cells of winter bees contained large numbers of secretory vesicles
that are probably stored until the spring.
Hassan and Khater (2006) in Egypt, indicated that the maximum
development of hypopharyngeal glands was achieved between 6-12
days for all diets used as pollen substitutes.

6. Chemical analysis of royal jelly


Weaver et al. (1968) identified three new compounds isolated
from the royal jelly of honeybees (Apis mellifera, L.) as; 8-
hydroxyoctanoic acid, 3-hydroxydeeanoic acid and a dextrorotatory
isomer of 3,10-dihydroxydecanoic acid.
Ewies (1974) stated that the fresh royal jelly had higher nitrogen
content than the stored one. He added that the amino acid percentages
in the royal jelly were; Aspartic (3.95%), Serine (1.41%), Glutamic
(3.62%), Glycine (1.59%), Alanine (2.09%), Arginine (3.32%),
Isoleucine (5.22%) and Histidine (1.81%).
Standifer and Mills (1977) in USA, fed nurse bees in
experimental colonies alternately with two diets; (1) a balanced diet
containing soybean flour, wheat, pulverized meat scraps, bee-collected
pollens, sucrose and water, and (2) an all- carbohydrate diet of mixed
sugars and water. The larval food (royal jelly) produced by the bee was
analyzed full antithetic acid, pyridoxine, niacin and ascorbic acid. The
amount of larval food produced by bees decreased with age regardless
of diet, its content of antithetic acid, pyridoxine and niacin. Differences

36
in the amount of ascorbic acid may be due to seasonal or metabolic
factor.
Lercker et al. (1981) characterized the fatty acid constituents of
the lipid fraction of royal jelly. Among the organic acids found after
fractionation by thin layer chromatography of the corresponding methyl
esters, the following compounds were identified by combined GC-MS;
saturated and unsaturated linear fatty acids, saturated and unsaturated
linear and branched dicarboxylic acids and mono and dihydroxy acids.
The most common characteristic of the organic acids was that most
contained 8 or 10 carbon atoms, whether saturated or unsaturated,
linear or branched.
Chiron (1982) described a new synthesis of (E)-10-hydroxy-2-
decenoic acid (royal jelly acid), starting from 8-bromo-l-octanol via
condensation of malonic acid with 8-hydroxyoctanal or 8-
acetoxyoctanal.
Karaali et al. (1988) in Turkey,characterized the composition of
Turkish royal jelly as determined from a sample provided by a local
commercial producer. The constituents analyzed were; moisture, total
sugars, lipids, proteins, amino acids, ash, minerals (iron and calcium)
and vitamins (thiamin, riboflavin and niacin). The same royal jelly
sample was freeze-dried, both at a laboratory and an industrial scale.
The compositions of freeze-dried products were investigated
comparatively through determination of total acidity over a 6-months
period. Various formulations were developed for tablet-making and the
most favorable formulation was recommended for industrial
production.

37
Laidlaw (1992) in the book of " The hive and the honeybee"
determined the chemical composition of royal jelly as it had the
following averages: moisture, 66.05%; protein, 12.34%; total reducing
substance 12.49%; total lipids, 5.46%; ash, 0.82% and undetermined
substances 2.84%.
Jia et al. (1995) described the application of capillary
electrophoresis (CE) to the separation and determination of the active
ingredient, 10-hydroxy-2-decenoic acid, in royal jelly with direct on-
column UV detection at 214 nm. using a cathodic injection and anodic
detection scheme. 10- hydroxy-2-decenoic acid (10-HDA) was
separated and detected in less than 10 min in a fused silica capillary
column with a phosphate buffer at pH 7.3 with an applied voltage of 20
KV followed by direct UV detection. The use of
cetyltrimethylammonium bromide (CTAB) as electroosmotic flow
modifier allows the rapid separation of 10-HDA from other
constituents in royal jelly by reversing the direction of electroosmotic
flow. The influence of organic solvents in the electrolyte on separation
selectivity was also discussed.
Krell (1996) in the United Nations, stated that the analysis of
royal jelly was generally based on the quantitative determination of the
three principal categories of compounds (lipids, sugars and proteins).
Lipids were the most important compounds in determining the
authenticity or adulteration of royal jelly, since several of them were
not found in any other natural products. The qualitative and
quantitative analysis of the lipid fraction also makes it possible to
determine the amount of royal jelly in a multi-component product.

38
Among the biologically active components, the vitamin content could
give an indication of the corresponding (assumed biological) activity of
royal jelly.
Schmitzova et al. (1998) reported that characterization of
major proteins of honeybee larval jelly (49–87 kDa) was performed by
the sequencing of new complementary DNAs (cDNAs obtained from a
honeybee head cDNA library), by the determination of N-terminal
sequences of the proteins, and by analyses of the newly obtained and
known sequence data concerning the proteins. It was found that royal
jelly (RJ) and worker jelly (WJ) contain identical major proteins and
that all the proteins belong to one protein family designated MRJP
(from major royal jelly proteins) and it contained a relatively high
amount of essential amino acids. These findings support the idea that
MRJPs play an important role in honeybee nutrition.
Albert et al. (1999) isolated and sequenced cDNA encoding a
new member of the gene family of major royal jelly proteins (MRJPs)
from the honeybee, Apis mellifera. They found that the MRJPs
appeared to have evolved a novel nutritional function in the honeybee.
Genc and Aslan (1999) in their research analyzed several royal
jellies and commercial products containing royal jelly for their trans-
10-hydroxy-2-decenoic acid (10-HDA) content by using a column
liquid chromatography technique. Ten samples claimed to be pure royal
jelly, containing 10-HDA between 0.75 and 2.54%. Seven samples
claimed to contain royal jelly as an ingredient which ranged from non-
detectable to 0.054%. The technique was found to be rapid with high
recovery.

39
Marconi et al. (2002) analyzed 15 samples of royal jelly for
protein, lysine and furosine content. In addition a commercial sample
of royal jelly at the beginning of its commercial shelf life was stored at
both of 4°C and room temperature to assess the development of Millard
reaction (furosine). The samples which came from known origin had a
significant lower content of furosine when compared to those of
imported ones. The average content of furosine increased in the
samples stored at room temperature by a higher level than those stored
at 4°C for the same period.
Boselli et al. (2003) identified free amino acids (FAAs) in royal
jelly (RJ) with mass spectrometric detection (GC-MS). The total FAA
content was 7.3 mg/g RJ on average; the major FAAs were proline,
lysine, glutamate, β-alanine, phenylalanine, aspartate and serine. The
FAA fraction was monitored in RJ frozen immediately after sample
collection (the control) and in aliquots of the same sample stored at two
different temperatures (room temperature and 4 °C) for different time
intervals (3, 6 and 10 months). The FAA content was constant
throughout the storage at 4 °C. At room temperature, proline and lysine
increased after three months to 6.8 and 3 mg/g, and then decreased after
6 and 10 months to 3 and 1 mg/g, respectively.
Egyptian organization for standardization and quality (2005)
determined the specifications of fresh and powdered royal jelly. As for
the fresh royal jelly it's content percentages should be as follow:
moisture not more than 66%, protein not less than 12%, 10 hydroxy-2-
decenoic acid not less than 2%, lipids not less than 5.5%, carbohydrates
not less than 12% and ash not more than 0.8%. In the case of powdered

40
royal jelly the percentages of it's content should be as follow: moisture
not more than 5%, protein not less than 32%, 10 hydroxy-2-decenoic
acid not less than 6%, arsenic 1ppm, copper 1ppm and must be free of
pathogenic germs and pesticides.
Messia et al. (2005) determined the level of furosine in the royal
jelly and showed that the Millard reaction was already occurred at the
hives as the furosine level was increased gradually from 1, 2 and 3 days
after grafting. The freeze-dried royal jelly was more pone to early stage
of Millard reaction than fresh one.
Melliou and Chinou (2005) isolated 7 new compounds from the
royal jelly and it was the first time to isolate these compounds as
natural products. The 7 compounds were; fatty acids: 10-
acetoxydecanoic acid, trans-10-acetoxydec-2-enoic acid, 11-
oxododecanoic acid, (11S)-hydroxydodecanoic acid, (10 R, 11R)-
dihydroxydodecanoic acid, 3, 11-dihydroxydodecanoic acid and (11S),
12-dihydroxydodecanoic acid. The structures of these compounds were
determined by spectroscopic methods.
Noda et al. (2005) in Japan, characterized novel fatty acids in
the royal jelly of honeybees (Apis mellifera). TLC analysis showed that
the chloroform/methanol extract obtained from royal jelly consists
mainly of fatty acids. By means of reversed-phase HPLC with various
solvent systems, 13 compounds were obtained in pure state. Their
structures including absolute configurations were determined by
chemical, NMR, and MS spectral analysis. Six compounds were
identified as novel mono- or diesters of 10-hydroxy-2E-decenoic acid
in which the hydroxyl group was esterified by another fatty acids unit,

41
and one was hydroxy- 2E-decenoic acid 10-phosphate. In addition, they
demonstrated that 9-hydroxy-2E-decenoic acid exists as a mixture of
optical isomers.
Qu et al.(2007) analyzed in detail and compared proteins
contained in royal jelly (RJ) derived from Chinese and European
honeybees. They found remarkable differences in the heterogeneity of
major royal jelly proteins (MRJPs), MRJP2 and MRJP3, in terms of
molecular weight and isoelectric points between the two species of RJ.
MRJP2 and MRJP3 produced by the Chinese honeybee were less
polymorphic than those produced by the European honeybee.
Schönleben et al. (2007) applied different methods for the pre-
fractionation and separation of royal jelly proteins in order to
circumvent the shortcomings of the individual techniques and achieve a
high coverage of the royal jelly proteome. They were able to identify
20 different proteins in total, as well as to show a very high degree of
cleavage of different proteins of the major royal jelly protein family.
Furthermore, they investigated the protein phosphorylation of royal
jelly proteins, and identified and located two phosphorylation sites
within venom protein 2.
Zhou et al. (2007) used the ultra performance liquid
chromatography (UPLC) and high performance liquid chromatography
(HPLC) methods to detect the trans-10-hydroxy- 2-decenoic acid (10-
HDA) content in royal jelly cream and lyophilized powder. The two
methods were validated for the determination of practical royal jelly
products. The concentration of 10-HDA ranged from 1.26 to 2.25% for
pure royal jelly cream samples and 3.01–6.26% for royal jelly

42
lyophilized powder samples. For 30 royal jelly products, the 10-HDA
content varied from no detectable to 1.005%.
Ishmuratov et al. (2008) used two approaches for synthesis of
9-oxo- and 10-hydroxy-2E-decenoic acids, biologically active
components of queen substance and royal jelly of honeybees,
respectively, starting with allyl bromide and 1,7-octadiene and using
chemo- and regioselective transformations of the common intermediate
building block 7-octen-1-ylacetate.

43
MATERIAL AND METHODS
This work was carried out in the Apiary yard of the Agricultural
Experimental Station, Faculty of Agriculture, Cairo University, Giza
Governorate during two successive years of 2008 and 2009 to
investigate the following aims:
1. Evaluation of some factors affecting royal jelly production in
honeybee colonies
a. Preparing experimental colonies
Twenty four honeybee colonies were used representing the three
following hybrids
1. Egyptian race (Apis mellifera lamarkii L.).
2. Carniolian hybrid (Apis mellifera L.).
3. Italian hybrid (Apis mellifera L.).
4. Local Carniolian race (Apis mellifera carnica) which used
only as donor colonies.
Queens of Egyptian race were obtained from Manfalout region,
Assiout Governorate. While the queens of local Carniolian race were
obtained in the early spring of the year 2008. On the other hand, queens
of the Carniolian and Italian hybrids were obtained from the apiary
yard of the Agricultural Experimental Station, Faculty of Agriculture,
Cairo University in the early spring of the year 2008.
Six honeybee colonies were chosen for each race and hybrid.
These colonies were classified into two groups (3 colonies per each
group).
The first group was used as royal jelly production queenless
colonies (RJPQC). The second one was considered as donor colonies

44
(DC) which used for providing RJPQC with young larvae (24 hours
old). These colonies were also used to provide RJPQC with sealed
worker brood combs immediately before emergence in order to
increase young worker bees that secreting royal jelly with high quality
and quantity to feed grafted larvae in RJPQC.
The experimental colonies were exposed to routine work such as
feeding with sugar syrup (1 sugar:1 water) 3 times weekly and for
controlling Varroa mites using formic acid 60%.
b. Preparing royal jelly production queenless colonies (RJPQC)
Royal jelly production queenless colonies were used as starter
and finisher colonies. These colonies must be strong (very crowded
with bee workers), as shown in Fig.1. Preparation of RJPQC started 7-
10 days before the beginning of experiments in order to increase the
number of young workers and stimulate their hypopharyngeal and
mandibular glands which are responsible for producing royal jelly by
intensive feeding.
For preparing RJPQC, the following procedures were done;
Honeybee production colonies (RJPQC) were supplied with
sealed worker brood immediately before emergence to provide
production colonies with young worker bees from the donor colonies
(DC) with an average of 1-2 combs / week (Fig. 2). Colonies were
supplied with artificial feeding of sugar syrup (1 sugar:1 water) 3 times
weekly (Fig. 3). Then, the queen and young opened worker brood
combs (1-3 days old) were removed from the colony 24 hours before
grafting, subsequently bees were reserved on the least number of

45
combs, according to Abd Al-Fattah (1987). The grafting frame place
was determined (between two combs of food and old larvae).

Fig. 1. A comb covered with bees. Fig. 2. A sealed comb.

Fig. 3. Artificial feeding (sugar syrup).

c. Preparing the grafting frame


Hoffman frame with 3 wooden bars was used for grafting (Fig.
4). Each bar had 16 fixed queen wax cups (Sahinler and Sahinler,
2002).

46
Queen cup formation is a simple technique that can be
summarized as follows; bees wax (Fig. 5) was melted in a melting wax
apparatus (Fig. 6). Thereafter the queen cup forming stick (Fig. 7) was
dipped into cold water for a suitable time, then removed, shaken off,
and dipped in the melting bees wax for few seconds. This procedure
was repeated 2 times.

Fig. 4. Hoffman frame with 3 wooden Fig. 5. Bees wax.


bars, each had 16 fixed queen.
wax cups.
With hand fingers, each formed queen cup (Fig. 8) was twisted
and pulled out, and then the queen cup forming stick was dipped again
in the water before forming another cup. Hundreds of queen cups were
made during this experiment.
The following procedure used to fix the queen cups into the
wooden bar with melting bee wax;
A thin layer of bee wax was spilled on the wooden bar, and then
queen cups were fixed slightly on it, followed by spilling another thick
layer for fixing queen cups hardly on the bar. When wax layer was
hardened, bars were arranged in the grafting frame and used for

47
grafting. After harvesting royal jelly, queen cells removed from bars
using a sharp knife and melted in the wax melting apparatus to use
again.

Fig. 6. Melting wax apparatus. Fig. 7. Queen cups forming sticks.

Fig. 8. Formed queen cups.

d. Grafting technique
Grafting denotes transferring young larvae from opened brood
combs to grafting frames. In all experiments, grafting was done at 8.0
a.m. Each production cycle (experiment) lasted about 10-22 days using

48
4-7 grafting frames. This technique had several procedures that
summarized as follows:
Donor colony's queen was excluded on an empty marked wax
comb for 24 hours. After laying eggs, it removed and the comb was
kept in the colony till eggs hatch.
The empty grafting frame, which had the queen cups on its bars,
was situated in production colonies 24 hours before grafting as the
frame acquires colony's pheromone and nurse bees polish wax cups.
This procedure was responsible for high acceptance of grafted larvae.
In the next day, grafting and marked frames removed from
production and donor colonies, respectively and transferred to grafting
room using brood transferring box (Fig. 9).
A droplet of royal jelly solution (1 royal jelly:1 warm water)
was dropped in the bottom of a queen cup with a wooden tool. Then 24
hours old larvae were transferred from the marked comb to the queen
cups carefully using grafting needle (Figs 10 and 11) (Sahinler et al.,
1997; Khattab et al., 1998 and Sharaf El-Din et al., 2000).
The royal jelly production queenless colonies (RJPQC) divided
into two groups. In the first group of colonies, the genetic origin of
larvae grafted in the grafting frame varied from the genetic origin (non-
related grafted larvae) of the production colony. As for the second
group the genetic origin of larvae grafted in the grafting frame was as
same as the genetic origin (related grafted larvae) of the production
colony.

49
Fig. 9. Brood transferring box. Fig. 10. Grafting needles.

Fig. 11. Grafting needle in a queen cup.

1. Non related grafted larvae


Experiments of this study were conducted using Egyptian and
Carniolian hybrids queenless production colonies. Each hybrid was
grafted with different genetic origin larvae; viz.,
a. Egyptian race grafted with different genetic origin larvae
throughout the period extended from 22/3/2008 to 31/3/2008.

50
b. Carniolian hybrid grafted with Carniolian race throughout the
period extended from 21/4/2008 to 30/4/2008.
c. Carniolian hybrid grafted with different genetic origin larvae
throughout the period extended from 4/7/2008 to 19/7/2008.
d. Carniolian hybrid grafted with different genetic origin larvae
throughout the period extended from 6/8/2008 to 15/8/2008.
e. Egyptian race grafted with different genetic origin larvae
throughout the period extended from 22/3/2009 to 6/4/2009.
While in the second one, the genetic origin of larvae which were
grafted in the grafting frame were as same as the genetic origin (related
grafted larvae) of the production colony as follows
2. Related grafted larvae
This study was conducted using Egyptian, Italian and Carniolian
hybrids queenless production colonies which was used as production
and donor colonies as follows:
a. Egyptian race colonies used as production and donor colonies
throughout the period extended from 8/4/2009 to 20/4/2009.
b. Italian hybrid colonies used as production and donor colonies
throughout the period extended from 15/7/2009 to 2/8/2009.
c. Carniolian hybrid colonies used as production and donor
colonies throughout the period extended from 13/8/2009 to 28/8/2009.
Bars were arranged in grafting frame and then transferred
carefully from grafting room to production colony using brood
transferring box, and situated between two combs (food and old larvae)
then the colony was covered carefully. A new grafting frame was
grafted every 3 days till the end of experiment.

51
In the next day of grafting, the first grafting frame was observed
carefully for counting the number of accepted opened queen cells. The
same routine work was repeated daily.
In addition, royal jelly was harvested from the grafting frame
during the fourth day and a new frame was grafted, etc.
At the same time, RJPQC were supplied weekly with 1-2 sealed
worker brood combs immediately before emergence (from donor
colonies) to provide RJPQC with young worker nurse bees.
All experimental colonies were supplied with sugar syrup (1
sugar:1 water) 3 times weekly as a source of feeding.
e. Royal jelly harvesting
This process was done in grafting room 3 days after the grafting.
The harvesting process was done as follows;
Grafting frame was taken from the production colony and
transferred to grafting room using brood transferring box, then bars
were pulled from the grafting frame. The accepted larvae were
removed from the queen cells, and royal jelly was harvested using
wooden harvest tool (Fig. 12). Harvested royal jelly was filtered with
sterilized gauze to get rid of wax particles. Royal jelly harvested from
each bar was separated in previously weighted dark bottle which was
weighted again using electronic balance for 3 decimals numbers to
define the net weight of harvested royal jelly (Fig. 13). A dark bottle
containing harvested royal jelly was stored in a deep freezer for
chemical analysis.
The following measurements were recorded:
1. Percentage of accepted grafted larvae / bar / grafting frame.

52
2. Percentage of accepted grafted larvae / grafting frame.
3. Amount of harvested royal jelly / bar / grafting frame.
4. Amount of harvested royal jelly / grafting frame.

Fig. 12. The wooden harvest tool. Fig. 13. Different containers
for harvested royal
jelly.
f. Laying workers
Percentage of laying workers was estimated every 3 days during
the experiment period as follows;
The sample was chosen from the worker bees on the grafting
frame. Ten worker bees were obtained and examined for their ovaries
development according to (El-Enany 2006). The abdomen of the
worker was opened by making an incision along the lateral and anterior
sides of the abdomen, and then the bees were pinned on a wax dish and
submerged in buffer solution under a binocular. The development of
ovaries was determined.
The results were calculated as a mean every 3 days for each
harvesting date/colony.

53
g. Number of combs covered with bees (colony strength).
Number of combs covered with bees (C.S.) in the production
colonies were counted every 3 days and recorded.
h. Seasons
Experiments were carried out in late spring and summer seasons
of 2008 and 2009. Each experiment lasted about 8-18 days before the
appearance of laying workers. All experiments were repeated 3 times.
i. Statistical analysis
The SPSS computer program did statistical analysis.
Data obtained from the experiments were analyzed using the
analysis of variance (ANOVA) and the least significant difference test
(L.S.D.).
Simple correlation (r) and simple regression (b) were calculated
to obtain information about the direct (primary) effect of:
a. Mean percentage of accepted grafted larvae on mean amount
of harvested royal jelly / harvesting date.
b. Mean percentage of laying workers on both of mean
percentage of accepted grafted larvae and mean amount of harvested
royal jelly / harvesting date.
c. Mean number of combs covered with bees on both of mean
percentage of accepted grafted larvae and mean amount of harvested
royal jelly / harvesting date.
2. Determination of amino and fatty acid percentages in samples
of the produced royal jelly
This part of research included analysis of royal jelly samples for
determination type and percentage of amino and fatty acids.

54
This study was carried out in the Faculty of Agriculture
Research Park (FARP). Eleven samples of the produced royal jelly
represented the related colonies (grafted larvae had the same genetic
origin of the production colonies) for the Egyptian race, Italian hybrid
and Carniolian hybrid were used and analyzed to determine the
percentage of amino (essential and non essential) and fatty acids. The
tested samples gained from different harvesting numbers under
different percentages of laying workers for the production colonies.
Three replications were maintained for each sample.
a. Determination of amino acid percentages
The method of Block et al. (1958) was used to determine the
amino acids in the royal jelly samples using the amino acid analyzer
apparatus (Automatic Amino Acid Analyzer, AAA400, INGOS Ltd).
Obtained results are expressed as gm/100 gm of RJ.
b. Determination of fatty acid percentages
For the determination of the fatty acids percentages in the tested
samples of RJ, the method reported by Farag et al. (1986) and
A.O.A.C. (2000) using gas liquid chromatography (GLC) was done and
the results were expressed as gm/100 gm of RJ.

55
RESULTS AND DISCUSSION
The production of royal jelly from honeybee colonies has
currently spread in most apiaries in different governorates of Egypt.

1. Evaluation of some factors affecting royal jelly production in


honeybee colonies.

The different factors influencing royal jelly production were


investigated under field conditions in the apiary yard of the
Agricultural of Experimental Station, Faculty of Agriculture, Cairo
University, Giza Governorate in the late spring and summer of 2008
and 2009. These factors included genetic origin of honeybee,
harvesting date, bar position, laying workers and number of combs
covered with bees (colony strength).

a. Non related grafted larvae


Inspection of bars of each studied race and hybrid was done
each 3 days and continued till the appearance of laying workers, thus
number of harvesting dates varied.

1. Egyptian race grafted with different genetic origin larvae


throughout the period extended from 22/3/2008 to 31/3/2008
In this study, the mean percentages of accepted grafted larvae
and harvested royal jelly amounts (gm) in bars for Egyptian race
production colonies grafted with different genetic origin larvae were
estimated each 3 days.

a. Acceptance percentage of grafted larvae


Table (1) presents mean percentages of accepted grafted larvae
and harvested royal jelly amounts (gm) in bars for Egyptian race

56
production colonies which grafted with different genetic origin larvae
throughout the period extended from 22/3/2008 to 31/3/2008.
In the first harvesting date, the data illustrated in Table (1)
indicated that the acceptance percentages of grafted larvae in the three
bars of the first production frame were highly significant. The highest
percentage of acceptance was gained at the second bar (Carniolian
race) (87.5%) followed by the first one (Italian hybrid) (70.8%)
whereas the third bar had the lowest percentage (Egyptian race) (8.3%).
On the contrary, there were no differences between the three
bars of the second production frame during the second harvesting date.
As for the third date of harvesting, the statistical analysis (F test)
revealed significant differences between the three bars of the third
frame. The highest percentage of acceptance took place at the first bar
(Egyptian race) (91.7%) followed by the third (Carniolian race) then
the second (Italian hybrid) ones (79.2% and 62.5%, respectively).
Highly significant differences were obtained among the three
bars of the fourth production frame (Egyptian race) during the fourth
harvesting date (March, 31, 2008). The maximum percentage was
occurred at the first bar followed by the second then the third ones
(89.6%, 64.6% and 8.3%, respectively).
Results concerning mean percentages of accepted grafted larvae
(%) and harvested royal jelly amounts (gm) in harvesting dates for
Egyptian race production colonies which grafted with different genetic
origin larvae throughout the period extended from 22/3/2008 to
31/3/2008 are shown in Table (2) and Fig. (14). Mean

57
Table 1. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in bars for Egyptian race
production colonies grafted with different genetic origin larvae throughout the period from 22/3/2008 to
31/3/2008.
production frame

Genetic origin of
Harvesting date

Accepted grafted larvae (%) /race Harvested royal jelly amount

grafted larvae

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/

(hybrid)/bar (gm)/race(hybrid)/bar

Mean F value LSD 0.05 Mean F value LSD 0.05

Italian hybrid 16 70.8 b 0.731 a


1st **
3 Carniolian race 16 87.5 a 46.17 10.1 0.412 b 28.06 ** 0.202
58

22/3/2008
Egyptian race 16 8.3 c 0.030 c
Carniolian race 16 16.7 0.086
2nd
3 Egyptian race 16 16.7 1.68 ns 0.202 1.91 ns 0.15
25/3/2008
Italian hybrid 16 27.1 0.559
Egyptian race 16 91.7 a 0.865 a
3rd
3 Italian hybrid 16 62.5 c 10.65 * 3.5 0.460 c 15.74** 0.142
28/3/2008
Carniolian race 16 79.2 b 0.624 b
89.6 a 0.911 a
4th
3 Egyptian race 48 64.6 b 66.52 ** 9.1 0.340 b 22.67 ** 0.303
31/3/2008
8.3 c 0.032 c
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

58
percentages of acceptance varied between 20.1 and 77.8%. Statistically,
highly significant differences were obtained between the mean
acceptance percentages of grafted larvae at different harvesting dates.
Highly percentage of accepted larvae was gained in the third harvesting
date (77.8%), followed by the first and fourth ones in which 55.6 and
54.2%, respectively were recorded. On the other hand, the lowest value
was recorded in the second harvesting date (20.1%).
Table (3) demonstrates mean percentages of accepted grafted
larvae and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Egyptian race production colonies which grafted
with different genetic origin larvae throughout the period extended
from 22/3/2008 to 31/3/2008.
When considering the three bars, highly significant differences
attained between the four harvesting dates.
In the first bar, the highest percentages of acceptance were
91.7% and 89.6% gained at the third and fourth harvesting dates,
respectively, followed by the first harvesting date which recorded
70.8%. The second one came in the last rank by having the lowest
value (16.7%).
Considering the second bar, the chief percentage of acceptance
took place in the first harvesting date (87.5%), followed by the fourth
and third harvesting dates which recorded 64.6% and 62.5% then the
second date that ranked the last one (16.7%).
As for the third bar, the maximum percentage of acceptance was
obtained at the third harvesting date (79.2%), followed by the second

59
Table 2. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) harvesting dates for
Egyptian race production colonies grafted with different genetic origin larvae throughout the period
from 22/3/2008 to 31/3/2008.

Genetic origin of
Harvesting date

grafted larvae

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/
production

Harvested royal jelly amount


frame
Accepted grafted larvae (%) /race
(gm)/race(hybrid)/harvesting
(hybrid)/harvesting date
date

Italian hybrid 16
1st 16
3 Carniolian race 55.6 b 0.391 b
22/3/2008
Egyptian race 16
60

Carniolian race 16
2nd
3 Egyptian race 16 20.1 c 0.282 c
25/3/2008
Italian hybrid 16
Egyptian race 16
3rd 16
3 Italian hybrid 77.8 a 0.650 a
28/3/2008
Carniolian race 16
4th
3 Egyptian race 48 54.2 b 0.428 b
31/3/2008
General mean 51.9 0.438
F value 5.51 * 7.60**
LSD 0.05 2.2 0.048
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

59
80 0.8
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


70 Harvested royal jelly 0.7
larvae
larvae %

60 0.6
grafted
grafted

50 0.5
of accepted
Meanof%accepted

40 0.4

61
30 0.3
Mean

20 0.2

10 0.1

0 0
22/03/2008 25/03/2008 28/03/2008 31/03/2008
Harvesting dates

Fig. 14. Mean (%) Fig.of14. Means ofgrafted


accepted acceptedlarvae
graftedand
larvae percentages
harvested andjelly
royal harvested royal(gm)
amounts jelly amounts (gm) indates for
in harvesting
Egyptianharvesting
race queenless
dates forproduction colonies
Egyptian race grafted
queenless withcolonies
production different genetic
grafted withorigin larvae
different geneticthroughout
the period from
origin 22-3-2008
larvae to 31-3-2008.
throughout the period from 22-3-2008 to 31-3-2008.
60
Table 3. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Egyptian race production colonies grafted with different genetic origin larvae
throughout the period from 22/3/2008 to 31/3/2008.
production frame

Genetic origin of
Harvesting date

Harvested royal jelly amount

grafted larvae

No. of grafted
(hybrid) /bar
larvae / race
Accepted grafted larvae (%) /race
No. of bars/

(gm)/race(hybrid)/ bar/harvesting
(hybrid)/bar/harvesting date
date

No. of bar No. of bar


st
1 2nd 3rd
1st
2nd 3rd
Italian hybrid 16
1st 16
3 Carniolian race 70.8 b 87.5 a 8.3 c 0.731 c 0.412 a 0.030 b
22/3/2008 16
Egyptian race
62

Carniolian race 16
2nd
3 Egyptian race 16 16.7 c 16.7 c 27.1 b 0.086 d 0.202 c 0.559 a
25/3/2008
Italian hybrid 16
Egyptian race 16
3rd 16
3 Italian hybrid 91.7 a 62.5 b 79.2 a 0.865 b 0.460 a 0.624 a
28/3/2008 16
Carniolian race
4th
3 Egyptian race 48 89.6 a 64.6 b 8.3 c 0.911 a 0.340 b 0.032 b
31/3/2008
General mean 59.7 55.5 38.2 0.560 0.358 0.404
F value 51.25 ** 28.05 ** 33.23 ** 25.85 ** 6.50 * 4.68 *
LSD 0.05 8.0 7.2 3.1 0.038 0.066 0.091
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

61
one which recorded 27.1% while both the first and fourth ones had the
lowest same value (8.3%). In general, the mean percentage of
acceptance / colony was 51.9%.

b. Amount of harvested royal jelly


Results of mean harvested royal jelly amount (gm) in bars for
Egyptian race are tabulated in Table (1) which indicated that the
differences among harvested royal jelly in the three bars were
significantly or highly significant affected with the 1st , 2nd, 3rd and 4th
production frames.

Statistically in the first production frame, the highest amount of


harvested royal jelly (gm) was gained at the first bar (0.731 gm)
followed by the second one then the third bar (0.412 and 0.030 gm,
respectively).

On the other side, results of the second production frame bars


illustrated non significant differences between the three bars.

From the results of the third production frame, it could be


observed that the first bar gave the maximum amount of royal jelly
(0.865 gm) which was followed by the third one (0.624 gm) whereas
the least value was gained in the second bar which had 0.460 gm.
Results of the fourth production frame, statistically arrange the
harvested amounts of royal jelly that obtained from the three bars
descendingly as follows; the first bar, the second one and the third bar
in which 0.911, 0.340 and 0.032 gm were gained, respectively.
Based on mean amounts of harvested royal jelly for Egyptian
race as shown in Table (2) and graphically illustrated in Fig. (14), it

63
obviously appeared that there were highly significant differences
among the amounts of royal jelly throughout the four harvesting dates
which ranged between 0.282 gm and 0.650 gm. However, the highest
amount of harvested royal jelly (gm) was gained at the third harvesting
date (0.650 gm). The fourth and first harvesting dates ranked the
second order where 0.428 gm and 0.391 gm royal jelly, respectively
were harvested. The second harvesting date had the last rank in which
0.282 gm royal jelly was harvested.
Results obtained for the means of harvested royal jelly amounts
(gm) in each bar during harvesting dates for Egyptian race
demonstrated significant or highly significant variations among
harvested royal jelly from the each bar throughout the four harvesting
dates.
Results of the first bar revealed that the highest amount of
harvested royal jelly (gm) was got in the fourth harvesting date (0.911
gm), followed by the third date (0.865 gm) then the first one (0.731
gm) while the least amount (0.086 gm) was harvested in the second
harvesting date.
Results of the second bar, revealed that the highest amounts of
harvested royal jelly were attained in both the third and first harvesting
dates (0.460 gm and 0.412 gm, respectively). The fourth harvesting
date came in the second rank in which 0.340 gm was gained. While the
least amount (0.202 gm) was obtained in the second harvesting date.
In view of results of the third bar, it was obvious that the high
amounts of royal jelly were harvested in both the third and second
harvesting dates (0.624 gm and 0.559 gm, respectively). On the

64
contrary, the low amounts were recorded in both the fourth and first
ones where 0.032 gm and 0.030 gm, respectively were harvested.
While the mean amount of harvested royal jelly / colony was 0.438 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees on both acceptance percentage
and amount of harvested royal jelly
Table (4) gives the simple correlations (r) and regressions (b) of
effect of the accepted grafted larvae percentage on harvested royal jelly
amount (gm) as well as laying workers percentage and number of
combs covered with bees/ production colony on both acceptance
percentage and harvested royal jelly amount (gm) for Egyptian race
queenless production colonies which grafted with different genetic
origin larvae throughout the period extended from 22/3/2008 to
31/3/2008.
Considering the effect of accepted grafted larvae percentage on
harvested royal jelly amount (gm), the simple correlation and
regression values were insignificant (3.558).
In case of the effect of laying workers on both accepted grafted
larvae percentage and harvested royal jelly amounts, this factor proved
an insignificant “r” and “b” values.
Also, the direct effect of number of combs covered with bees/
production colony („r‟ and „b‟ value) on both accepted grafted larvae

65
Table 4. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Egyptian race queenless production colonies
grafted with different genetic origin larvae throughout the period from 22/3/2008 to 31/3/2008.

Accepted Laying workers Number of combs covered with bees


Accepted grafted larvae (%)
Harvested royal jelly amount
grafted
larvae on Accepted grafted Harvested
Accepted grafted larvae Harvested royal jelly
Harvesting date

harvested larvae royal jelly


royal jelly
(gm)

Mean percentage

Mean number
66

r b t r b t r b t r b t r b t

1st
55.6 0.391 40 3
22/3/2008
2nd
3.558 ns

0.529 ns

0.562 ns

0.504 ns

0.643 ns
20.1 0.282 45 3.2
0.929

0.006

0.350

0.464

0.369

0.003

0.336

0.005

0.414

0.938
25/3/2008
3rd
77.8 0.650 60 3.5
28/3/2008
4th
54.2 0.428 80 3.8
31/3/2008
ns
Non significant

66
percentage (%) and harvested royal jelly amounts (gm) was
insignificant.
Obtained results could be summarized as significant or highly
significant differences were calculated between acceptance means (%)
and amount means (gm) of RJ / colony among the different grafted bars
represented the tested races and hybrid of bees at the same harvesting
dates. As for the acceptance percentages, the highest one occurred at
the Carniolian race followed by the Italian hybrid then the Egyptian
race. As for, the amount of RJ production, the chief amount was
harvested from the Italian hybrid then the Carniolian one and finally
the Egyptian race.
Considering the three bars, it could be said that the first bar gave
the highest percentage of acceptance and amount of RJ. The second one
had an intermediate acceptance percentage and the lowest amount of
RJ. In the mean time, the percentage of acceptance for the third bar was
the lowest while the amount of RJ ranked the second order.
The mean percentage of acceptance for these colonies was
51.9%, whereas the mean amount of harvested royal jelly was 0.438
gm.
The influences of accepted grafted larvae percentage on amount
of harvested royal jelly as well as the effect of each laying workers
percentage and number of combs covered with bees (C.S.) on both
acceptance percentage and amount of harvested royal jelly were
insignificant.

67
2. Carniolian hybrid grafted with Carniolian race larvae
throughout the period from 21/4/2008 to 30/4/2008
The studies of accepted grafted larvae percentages and harvested
royal jelly amounts (gm) in bars for Carniolian hybrid production
colonies grafted with Carniolian race larvae were carried out once
every 3 days.

a. Acceptance percentage of grafted larvae


The results concerning mean percentages of accepted grafted
larvae and harvested royal jelly amounts (gm) in bars for Carniolian
hybrid production colonies which grafted with Carniolian race larvae
throughout the period elapsed from 21/4/2008 to 30/4/2008 are
tabulated in Table (5).
The statistical analysis revealed significant variations in
acceptance percentages of grafted larvae in the three bars of the first
production frame.
The main percentage of 81.3% took place in the first bar, while
the minimum one of 58.4% happened in the third bar. The second bar
had an intermediate rank in which 62.5% of grafted larvae were
accepted.
During the second harvesting date, the accepted grafted larvae
was higher in the second bar since the mean percentage was 70.9%,
whereas the respective percentage means were 62.5% and 56.3% in the
first and the third ones, respectively.

The maximum of accepted grafted larvae occurred in the third


bar during the third harvesting date, where the mean percentage was

68
62.5%, followed by the first and second ones in which 50.0% and
37.5%, respectively recorded.

The results of the fourth harvesting date, declared that the


highest mean percentage was 66.7% which took place in the first bar,
while the second and third bars came next in the same group recording
25.0% and 22.9%, respectively.
Results regarding mean percentages of accepted grafted larvae
and harvested royal jelly amounts (gm) in harvested dates for
Carniolian hybrid production colonies which grafted with Carniolian
race larvae throughout the period extended from 21/4/2008 to
30/4/2008 are tabulated in Table (6) and graphically illustrated in Fig.
(15). These results showed that the accepted grafted larvae means were
ranged from 38.2% in the fourth harvesting date to 67.4% in the first
one. However, it could be mentioned that the variations between means
of accepted grafted larvae gave an evidence of highly significant value.
The percentage means could be arranged as follows: the first
harvesting date gave the highest value (67.4%), which was followed
descendingly by the second one (63.2%), the third harvesting date
which recorded 50.0% and the last one was the fourth harvesting date
with 38.2%.
Mean percentages of accepted grafted larvae and harvested
royal jelly amounts (gm) in each bar during harvesting dates for
Carniolian hybrid production colonies which grafted with Carniolian
race larvae throughout the period extended from 21/4/2008 to
30/4/2008 are shown in Table (7). Results of the statistical analysis

69
Table 5. Means (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in bars for Carniolian
hybrid production colonies grafted with Carniolian race larvae throughout the period from 21/4/2008 to
30/4/2008.
production frame

Genetic origin of

larvae / race/bar
Harvesting date

Accepted grafted larvae (%) Harvested royal jelly amount

grafted larvae

No. of grafted
No. of bars/

/race/bar (gm)/race/bar

Mean F value LSD 0.05 Mean F value LSD 0.05

81.3 a 2.268 a
1st
3 Carniolian race 48 62.5 b 7.89* 3.5 1.362 b 12.16 **
0.148
21/4/2008
70

58.4 c 1.382 b
62.5 b 1.287 a
2nd
3 Carniolian race 48 70.9 a 6.95 * 5.1 0.935 c 16.22 **
0.184
24/4/2008
56.3 c 1.125 b
50.0 b 0.959 a
3rd
3 Carniolian race 48 37.5 c 12.00 ** 6.9 0.739 c 11.86 **
0.039
27/4/2008
62.5 a 0.914 b
66.7 a 1.254 a
4th
3 Carniolian race 48 25.0 b 24.77 ** 6.0 0.578 b 12.99 ** 0.505
30/4/2008
22.9 b 0.423 c
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

70
demonstrated highly significant variances between mean percentages of
accepted grafted larvae in the four harvesting dates of each bar.
In the first bar, the highest percentage of acceptance was gained
in the first harvesting date (81.3%), followed by the fourth one which
recorded 66.7%. The second harvesting date came in the third rank
(62.5%). The percentage obtained from the third harvesting date
represented the lowest value (50.0%).
Regarding the second bar, the four harvesting dates could be
arranged descendingly according to percentages of accepted grafted
larvae as follows; the second harvesting date gave the maximum
percentage of 70.9%, followed by the 1st, 3rd and 4th ones with 62.5%,
37.5% and 25.0%, respectively.
During the third bar, both the second and third harvesting dates
gave the maximum acceptance percentages (62.5% and 62.5%
respectively), followed by the first one which recorded 58.4%. The
fourth harvesting date came in the last rank by having 22.9%.
Meanwhile, the percentage mean of acceptance / colony was 54.7%.

b. Amount of harvested royal jelly


The results illustrated in Table (5) for harvested royal jelly
amounts (gm) in bars for Carniolian hybrid production colonies grafted
with Carniolian race clarified that the amounts of harvested royal jelly
were significant or highly significant affected by the number of bar in
each harvesting date.

It could be seen from results of the first production frame that


the highest amount of harvested royal jelly (gm) was gained in the first

71
Table 6. Means (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in harvesting dates for
Carniolian production colonies grafted with Carniolian race larvae throughout the period from
21/4/2008 to 30/4/2008.

Genetic origin

No. of grafted
larvae / race
No. of bars/
production
Harvesting

of grafted
larvae
frame Accepted grafted larvae (%) Harvested royal jelly amount
date

/bar
/race /harvesting date (gm)/race/ harvesting date

1st
3 Carniolian race 48 67.4 a 1.671 a
21/4/2008
72

2nd
3 Carniolian race 48 63.2 b 1.116 b
24/4/2008

3rd
3 Carniolian race 48 50.0 c 0.871 c
27/4/2008

4th
3 Carniolian race 48 38.2 d 0.752 d
30/4/2008
General mean 54.7 1.103
F value 28.66 ** 26.01 **
LSD 0.05 4.1 0.129
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

72
90 1.8
Accepted grafted larvae
80 Harvested royal jelly 1.6

Mean of harvested royal jelly amounts (gm)


% of accepted grafted larvae
Mean of accepted grafted larvae %

70 1.4

60 1.2

50 1

40 0.8

73
30 0.6
Mean

20 0.4

10 0.2

0 0
21/04/2008 24/04/2008 27/04/2008 30/04/2008
Harvesting dates

Fig. 15. Means (%) Means


Fig. 15. of accepted graftedgrafted
of accepted larvaelarvae
and harvested royal
percentages andjelly amounts
harvested (gm)jelly
royal in harvesting dates
amounts (gm) in for
Carniolian hybrid
harvesting datesqueenless production
for Carniolian hybridcolonies grafted
queenless with Carniolian
production race larvae
colonies grafted throughout the
with Carniolian
period from 21- 4-2008 to 30-4-2008.
race larvae throughout the period from 21- 4-2008 to 30-4-2008.
73
bar (2.268 gm) followed by both the third and second ones in which
1.382 gm and 1.362 gm, respectively were attained.
In the second production frame bar, the maximum amount
occurred in the first bar (1.287 gm) followed by the third then the
second ones (1.125 gm and 0.935 gm, respectively).

As for the third production frame, the trend of the three bars in
harvesting royal jelly has same of those took place in the second
harvesting date. The greatest amount was gained at the first bar (0.959
gm) followed by both the third and second ones (0.914 gm and 0.739
gm respectively).

During the fourth production frame, the highest amount of


harvested royal jelly was achieved in the first bar (1.254 gm) then the
second one (0.578 gm) and finally the third one (0.423 gm).

It appeared from data presented in Table (6) and Fig. (15) that
there were highly significant differences between harvested royal jelly
amount (gm) means in harvesting dates for Carniolian production
colonies which grafted with Carniolian race larvae.

However, the amount of harvested royal jelly could be arranged


as follows; the first harvesting date gave the highest mean amount
(1.671 gm) . This date was followed descendingly by the second one
which had 1.116 gm, the third one with 0.871 gm, and the fourth
harvesting date which gave 0.752 gm.
Analysis of variances concerning the harvested royal jelly
amounts in each bar for Carniolian hybrid production colonies which

74
Table 7. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in each bar during
harvesting dates for Carniolian hybrid production colonies grafted with Carniolian race larvae
throughout the period from 21/4/2008 to 30/4/2008.
production frame

Genetic origin of

larvae / race/bar
Harvesting date

grafted larvae

No. of grafted
Accepted grafted larvae (%) /race/ Amount of harvested royal jelly
No. of bars/

bar/harvesting date (gm)/ race/bar/harvesting date

No. of bar No. of bar


st
1 2nd 3rd
1st
2nd 3rd
st
1
3 Carniolian race 48 81.3 a 62.5 b 58.4 b 2.268 a 1.362 a 1.382 a
21/4/2008

75
2nd
3 Carniolian race 48 62.5 c 70.9 a 62.5 a 1.287 b 0.935 b 1.125 b
24/4/2008
3rd
3 Carniolian race 48 50.0 d 37.5 c 62.5 a 0.959 c 0.739 bc 0.914 b
27/4/2008
4th
3 Carniolian race 48 66.7 b 25.0 d 22.9 c 1.254 b 0.578 c 0.423 c
30/4/2008
General mean 65.1 48.9 51.5 1.400 0.903 0.961
F value 9.57 ** 19.10 ** 15.29 ** 39.10 ** 29.96 ** 43.81 **
LSD 0.05 4.0 8.2 3.3 0.135 0.231 0.228

** Highly significant at probability level 0.01


Means within a column followed by different letter are significantly or highly significant (LSD test).

75
grafted with Carniolian race larvae shown in Table (7), confirmed
highly significant differences between means of royal jelly amounts in
the four harvesting dates.

In the first bar, the highest amount of harvested royal jelly (gm)
was gained at the first harvesting date (2.268 gm), followed by both the
second and fourth ones (1.287 gm and 1.254 gm, respectively), while
the third harvesting date came last rank (0.959 gm).

As for the second bar, the first harvesting date came in the first
order where maximum mean royal jelly amount was harvested (1.362
gm), followed by the second one (0.935 gm), then the fourth one (0.578
gm). The third harvesting date has an intermediate rank between the
second and fourth ones where 0.739 gm royal jelly was obtained.

The results of the third bar, showed that the maximum royal
jelly amount of 1.382 gm was achieved in the first harvesting date,
while the minimum value was got in the fourth one (0.423 gm). Both
the second and third harvesting dates ranked in the same order
following the first harvesting date where 1.125 and 0.914 gm,
respectively were attained. At same time, the mean amount of
harvested royal jelly / colony was 1.103 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
Table (8) gives the simple correlations (r) and regressions (b) of
the effect of accepted grafted larvae percentage on harvested royal jelly
amount (gm) as well as laying workers percentage mean and mean

76
number of combs covered with bees/ production colony on both
acceptance percentage and harvested royal jelly amount (gm) for
Carniolian hybrid queenless production colonies which grafted with
Carniolian race larvae throughout the period extended from 21/4/2008
to 30/4/2008.

Results of the effect of accepted grafted larvae percentage on


harvested royal jelly (gm/production frame), pointed out that the
resulting (r) and (b) values were insignificant.

As for the effect of laying workers, "t" value clarified highly


significant negative value in the case of mean percentages of accepted
grafted larvae. On the other side, the same factor proved an
insignificant value in the mean amount of harvested royal jelly.

In case of the effect of mean numbers of combs covered with


bees, the simple correlation and regression coefficients were highly
significant negative in case of mean percentages of accepted grafted
larvae, whereas they were insignificant in the mean amount of
harvested royal jelly.
Also obtained results mentioned that there were significant or highly
significant differences between means of accepted grafted larvae
percentages and harvested royal jelly amounts (gm) in three bars
correspond to the tested race of bees in the same harvesting date.
According to the results of the three bars, it has been found that the
highest percentage of acceptance and amount of RJ was occurred in the
first bar followed by the second one. The third bar showed the
lowest percentage of acceptance and amount of RJ.

77
Table 8. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and amount of harvested royal jelly for Carniolian hybrid queenless production colonies
grafted with Carniolian race larvae throughout the period from 21/4/2008 to 30/4/2008.
Accepted Laying workers Number of combs covered with bees
Harvested royal jelly amount
Accepted grafted larvae (%)
grafted
larvae on Accepted grafted Harvested royal Accepted grafted Harvested royal
Harvesting date

harvested larvae jelly larvae jelly


royal jelly
(gm)

Mean percentage

Mean number
78

r b t r b t r b t r b t r b t

1st
67.4 1.671 8 5
21/4/2008
2nd
63.2 1.116 10 5.2
7.605**
2.551ns

1.810ns
-0.983

-0.540

-0.788

-0.013

-0.986

-0.023

-0.941

-0.717
8.37**

3.94ns
0.875
0.027

24/4//2008
3rd
50 0.871 30 5.5
27/4/2008
4th
38.2 0.752 60 5.7
30/4/2008
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01

78
The means of acceptance percentage and harvested royal jelly
amount recorded 54.7% and 1.103 gm, respectively.
As for the effect of accepted grafted larvae percentage on
amount of harvested royal jelly, the results showed that no significant
differences were observed . On the other side, the influences of each of
laying workers percentage and number of combs covered with bees
(C.S.) were highly significant for the accepted grafted larvae and
insignificant with the harvested royal jelly.

3. Carniolian hybrid grafted with different genetic origin larvae


throughout the period extended from 4/7/2008 to 19/7/2008
a. Acceptance percentage of grafted larvae
Results of analysis of variance illustrated in Table (9) for means
of accepted grafted larvae (%) in bars for Carniolian hybrid queenless
production colonies which grafted with different genetic origin showed
significant or highly significant differences between 3 bars of first,
third, fourth and sixth production frames. On the other hand, the
variations were insignificant between 3 bars of the second and fifth
harvesting dates.

In the first production frame, the highest percentages of


acceptance were gained in the second and third bars (85.5% and 83.4%,
respectively), while the lowest percentage of 58.4% was recorded in the
first bar.

During the third production frame, the maximum acceptance


percentage occurred in the second bar (36.6%), followed with by the

79
Table 9. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in bars for Carniolian
hybrid queenless production colonies grafted with different genetic origin larvae throughout the period
from 4/7/2008 to 19/7/2008.
Harvesting date

Genetic origin

No. of grafted
(hybrid) /bar
No. of bars/
Accepted grafted larvae (%) Harvested royal jelly amount

larvae / race
production

of grafted
larvae
/race (hybrid)/bar (gm)/race(hybrid)/bar
frame

Mean F value LSD 0.05 Mean F value LSD 0.05

1st 58.4 b 1.862 c


3 Carniolian hybrid 48 85.5 a 8.70 * 4.1 2.875 a 6.30 *
0.165
4/7/2008 83.4 a 2.678 b
2nd 2.1 0.041 c
2.63 ns *
80

Carniolian race 48 12.5 0.360 a 8.23 0.113


7/7/2008 3 10.5 0.158 b
3rd 10.4 c 0.044 c
3 Carniolian race 48 39.6 a 16.50 ** 2.9 0.404 a 27.34 **
0.121
10/7/2008 27.1 b 0.261 b
4th 25.0 c 1.096 c
3 Carniolian race 48 66.7 a 25.06 ** 7.5 1.473 b 42.68 **
0.205
13/7/2008 56.3 b 1.864 a
5th 89.6 1.822
3 Carniolian hybrid 48 72.9 2.90 ns 1.948 2.39 ns
16/7/2008 68.8 1.312
6th 45.9 b 1.039 a
3 Carniolian race 48 18.07 ** 7.7
79.2 a 16.87 **
0.735 b 0.125
19/7/2008 31.3 c 0.553 c
ns
Insignificant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).
80
third one (27.1%), whereas the first bar represented the minimum
percentage (10.4% ).
The acceptance percentages in the fourth production frame had
the same trend of that gained in the third production frame. The
respective percentages were 66.7, 56.3 and 25.0% that happened in the
2nd, 3rd and 1st bars, respectively.

The highest percentage of acceptance for the sixth production


frame bars was gained in the second bar (79.2%), followed by the first
one (45.9%) and finally the third one (31.3%).

Means of accepted grafted larvae in harvesting dates for


Carniolian hybrid queenless production colonies which grafted with
different genetic origin larvae throughout the period extended from
4/7/2008 to 19/7/2008 shown in Table (10) and illustrated in Fig. (16)
revealed that the acceptance percentages varied from 8.4 to 77.1%. The
acceptance percentages were highly significant affected with harvesting
dates. Significantly, high percentages were recorded in both the fifth
and first harvesting dates (77.1% and 75.7% respectively), followed by
52.1% and 49.3% (in both the six and fourth ones, respectively, then
25.7% in the third one. The second harvesting date represented the last
rank of 8.4%.
Table (11) showed means of accepted grafted larvae percentages
(%) and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Carniolian hybrid queenless production colonies
which grafted with different genetic origin larvae, throughout the
period extended from 4/7/2008 to 19/7/2008.

81
Table 10. Mean(%) of accepted grafted larvae and harvested royal jelly (gm) amounts in harvesting dates for
Carniolian hybrid queenless production colonies grafted with different genetic origin larvae
throughout the period from 4/7/2008 to 19/7/2008.

Genetic origin

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/
Harvesting

production

of grafted
Harvested royal jelly amount

larvae
frame
date

Accepted grafted larvae (%)


(gm))/ race (hybrid)/ harvesting
/race (hybrid)/harvesting date
date

1st
3 Carniolian hybrid 48 75.7 a 2.472 a
4/7/2008
2nd
82

Carniolian race 48 8.4 d 0.187 e


7/7/2008 3
3rd
Carniolian race 48 25.7 c 0.237 e
10/7/2008 3
4th
3 Carniolian race 48 49.3 b 1.478 c
13/7/2008
5th
3 Carniolian hybrid 48 77.1 a 1.694 b
16/7/2008
6th
3 Carniolian race 48 52.1 b 0.776 d
19/7/2008
General mean 48.1 1.141
F value 35.43 ** 42.37 **
LSD 0.05 11.5 0.205
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

82
90 2.7
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


80 Harvested royal jelly 2.4
%
larvae
larvae

70 2.1
grafted

60 1.8
grafted

50 1.5
accepted
accepted

40 1.2

83
of of

30 0.9
Mean%
Mean

20 0.6

10 0.3

0 0
04/07/2008 07/07/2008 10/07/2008 13/07/2008 16/07/2008 19/07/2008
Harvesting dates

Fig. 16 Means of accepted grafted larvae percentages and harvested royal jelly amounts (gm) in
Fig. 16 Meanharvesting
(%) of accepted
dates forgrafted larvae
Carniolian andqueenless
hybrid harvestedproduction
royal jelly amounts
colonies (gm)with
grafted in harvesting
different dates for
Carniolian hybrid queenless production colonies grafted with different genetic
genetic origin larvae throughout the period from 7- 4-2008 to 19-4-2008. origin larvae throughout
the period from 4-7-2008 to 19-7-2008.
83
Table 11. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in each bar during
harvesting dates for Carniolian hybrid queenless production colonies grafted with different genetic
origin larvae, throughout the period from 4/7/2008 to 19/7/2008.

Genetic origin

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/

Accepted grafted larvae (%) /race Harvested royal jelly amount (gm)/
production
Harvesting

of grafted
larvae
frame

(hybrid)/bar/harvesting date race (hybrid)/ bar/harvesting date


date

No. of bar No. of bar


st
1 2nd 3 rd
1st
2nd 3rd
1st
3 Carniolian hybrid 48 58.4 b 85.5 a 83.4 a 1.862 a 2.875 a 2.678 a
4/7/2008
2nd
Carniolian race 48 2.1 f 12.5 g 10.5 e 0.041 c 0.360 e 0.158 e
84

7/7/2008 3
3rd
Carniolian race 48 10.4 e 39.6 f 27.1 d 0.044 c 0.404 e 0.261 e
10/7/2008 3
4th
3 Carniolian race 48 25.0 d 66.7 de 56.3 c 1.096 b 1.473 c 1.864 b
13/7/2008
5th
3 Carniolian hybrid 48 89.6 a 72.9 bce 68.8 b 1.822 a 1.948 b 1.312 c
16/7/2008
6th
3 Carniolian race 48 45.9 c 79.2 ac 31.3 d 1.039 b 0.735 d 0.553 d
19/7/2008
General mean 38.6 59.4 46.2 0.984 1.299 1.004
** ** ** ** **
F value 64.22 33.69 23.70 32.35 26.35 25.49**
LSD 0.05 7.3 8.2 7.2 0.094 0.243 0.124
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

84
Results of the third bar revealed that the highest percentage of
acceptance was gained in the first harvesting date (83.4%), followed by
the fifth one (68.8%) then the fourth date (56.3%). The sixth and third
harvesting dates came in the fourth rank (31.3 and 27.1%,
respectively), whereas the second harvesting date was ranked the last
order in which the lowest value was obtained (10.5%).

b. Amount of harvested royal jelly


Data illustrated in Table (9) revealed that the mean harvested
royal jelly amounts (gm/ bar) were significant or highly significant
affected with three bars of Carniolian hybrid colonies in all harvesting
dates with the exception of the fifth date that gave an insignificant
value. In those cases of significant or highly significant values in the
first, second and third dates, the highest harvested royal jelly was
gained in the second bar (2.875, 0.360 and 0.404 gm, respectively)
followed by the third one (2.678, 0.158 and 0.261 gm, respectively)
then the first bar (1.862, 0.041 and 0.044 gm, respectively).

As for the fourth date, the first bar recorded the lowest amount
of harvested royal jelly (1.096 gm), while the third bar reached the
maximum value (1.864 gm) followed by the second one (1.473 gm).

The last harvesting date (the sixth) had the trend of first, second
and third bars, respectively where they reached the amount of 1.039,
0.7335 and 0.553 gm, respectively.

Regarding the harvesting dates, the analysis of variance proved


highly significant differences between royal jelly amounts in the
different harvesting dates. The six dates could be arranged

85
descendingly according to their obtained amounts as follows; the first
harvesting date gave the highest amount (2.472 gm) when compared
with those obtained in other dates. The fourth harvesting date came in
the second rank (1.694 gm) followed by the fourth date in which 1.478
gm of royal jelly was harvested. The sixth date ranked the fourth order
(0.776 gm). The third and second harvesting dates ranked the fifth
order (0.237 gm and 0.187 gm, respectively).
Data concerning with the harvested royal jelly amounts in each
bar (gm/bar) during harvesting dates - shown in Table (15) - indicated
that there were highly significant differences between harvested royal
jelly amounts in the different harvesting dates for each bar.

In the first bar, the maximum amount of harvested royal jelly


(gm) gained in both the first and fifth dates (1.862 and 1.822 gm,
respectively), followed by both the fourth and sixth dates (1.096 and
1.039 gm, respectively. The third and second gained harvesting dates
ranked the last order, where relatively low amounts were gained (0.044
and 0.041 gm, respectively).

Harvesting dates of the second bar could be arranged in


descending orders according to their harvesting royal jelly amounts as
follows; the 1st, 5th, 4th, 6th and both the 3rd and 2nd dates in which
2.875, 1.948, 1.473, 0.735, 0.404 and 0.360 gm of royal jelly were
attained, respectively

Obtained results of the third bar indicated that the highest


amount of harvested royal jelly (gm) was gained in the first harvesting
date (2.678 gm), followed descendingly by the fourth, fifth and sixth

86
ones (1.864, 1.312 and 0.553 gm, respectively). Both the third and
second dates gave the least amounts, where 0.261gm and 0.158 gm
were harvested, respectively. Meanwhile, the mean amount of
harvested royal jelly / colony was 1.141 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
The results in Table (12) showed the simple correlations (r) and
regressions (b) of the effect of accepted grafted larvae percentage on
harvested royal jelly amount (gm) as well as laying workers percentage
mean and mean number of combs covered with bees/ production
colony on both acceptance percentage and harvested royal jelly amount
(gm) for Carniolian hybrid queenless production colonies grafted with
different genetic origin larvae throughout the period extended from
4/7/2008 to 19/7/2008.
The calculated effect of accepted grafted larvae mean
percentages on harvested royal jelly mean amounts (gm) indicated that
the effect of mean grafted larvae (%) was significantly positive (t =
4.067) on the mean amount of royal jelly.
As for the effect of laying workers, the direct relation between
this factor and both accepted grafted larvae percentages and harvested
royal jelly amounts - as calculated by simple regression values - were
insignificant.
Also, when concerning the calculated "t" values for the effect of
number of combs covered with bees, the results indicated that this
factor proved insignificant effects on both accepted grafted larvae

87
Table 12. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Carniolian hybrid queenless production colonies
grafted with different genetic origin larvae throughout the period extended from 4/7/2008 to 19/7/2008.
Accepted Laying workers Number of combs covered with bees
Harvested royal jelly
Accepted grafted

grafted larvae
Harvesting date

on harvested Accepted grafted Harvested royal Accepted grafted Harvested royal


amount (gm)
larvae (%)

royal jelly larvae jelly larvae jelly

percentage

number
Mean

Mean
r b t r b t r b t r b t r b t

1st
88

4/7/2008 75.7 2.472 20 5


2nd
7/7/2008 8.4 0.187 25 5.3
3rd
10/7/2008 25.7 0.237 45 5.6

0.519ns

0.280ns

0.450ns

0.327ns
-0.161

-0.101
4.067*
0.897

0.030

0.251

0.374

0.220

0.005
-0.14

-0.01
4th
13/7/2008 49.3 1.478 50 5.9
5th
16/7/2008 77.1 1.694 60 6.2
6th
19/7/2008 52.1 0.776 65 6.5
ns
Insignificant
* Significant at probability level of 0.05

88
percentages and harvested royal jelly amounts.

Also obtained results cleared that the variations among each of


the acceptance means (%) and the collected amount means of RJ (gm) /
colony were significant or highly significant among the different
grafted bars characterized the tested race and hybrid of bees at the same
harvesting date. Considering the three bars, it can be generally stated
that the first bar gave the minimum percentage of acceptance and
amount of RJ. In the meantime, the second bar had the maximum
percentage and amount. The third one recorded an in-between
percentage of acceptance and the amount of RJ.
The general mean of acceptance percentage for these colonies
was 48.1%, while general mean amount of harvested royal jelly was
1.141 gm.

Analysis of variance showed significant effect of the accepted


grafted larvae percentage on the amount of harvested royal jelly.
Reversely, the effect of laying workers percentage and number of
combs covered with bees (C.S.) on both acceptance percentage and
amount of harvested royal jelly were insignificant.

4. Carniolian hybrid grafted with different genetic origin


larvae throughout the period extended from 6/8/2008 to 15/8/2008
a. Acceptance percentage of grafted larvae
Table (13) presents the mean acceptance percentages of grafted
larvae in bars for Carniolian hybrid queenless production colonies
grafted with different genetic origin larvae throughout the period
extended from 6/8/2008 to 15/8/2008. the data proved that there were

89
Table 13. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in bars for Carniolian
hybrid queenless production colonies -grafted with different genetic origin larvae, throughout the
period extended from 6/8/2008 to 15/8/2008.
production frame

Genetic origin of
Harvesting date

grafted larvae

No. of grafted
Accepted grafted larvae (%) /race Harvested royal jelly amount (gm)

(hybrid) /bar
larvae / race
No. of bars/

(hybrid)/bar /race(hybrid)/bar

Mean F value LSD 0.05 Mean F value LSD 0.05

31.3 c 0.745 c
1st
3 Carniolian race 48 54.2 b 42.67 ** 10.5 1.222 b 21.36 ** 0.134
90

6/8/2008
87.5 a 1.396 a
39.6 c 0.671 c
2nd
3 Carniolian race 48 66.7 a 6.08 * 6.4 1.444 a 8.42 * 0.121
9/8/2008
52.1 b 1.179 b
66.7 b 0.515 b
3rd Carniolian
3 48 83.4 a 7.06 * 9.5 1.073 a 23.82 ** 0.069
12/8/2008 hybrid
54.2 c 0.419 c
45.9 b 0.497 b
4th
3 Carniolian race 48 77.1 a 23.41 ** 8.8 0.133 c 20.80 ** 0.039
15/8/2008
33.4 c 0.544 a

* Significant at probability level of 0.05


** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

90
significant or highly significant differences among mean acceptance
percentages of the three bars for each production frame.

As for the first production frame, the 3rd bar presented maximum
acceptance percentage (87.5%) then the second one (54.2%), while the
first bar represented the last category (31.3%). As regarding the second
frame, higher acceptance percentage took place in the second bar
(66.7%) followed by the third one (52.1%) then the first bar (39.6%).

The trend of acceptance grafted larvae percentages in three bars


of both the third and fourth production frames were the same. The
highest percentage was reached in the second bar (83.4 and 77.1%,
respectively) which followed by the first one (66.7 and 45.9%,
respectively) then the third bar (54.2 and 33.4%, respectively).

As regard of the results related to accepted grafted larvae


percentages throughout harvested dates (Table (14) and Fig. (17)), the
statistical analysis revealed significant variations between means of
larvae acceptance percentages. In this respect, the highest percentage of
acceptance was gained in the third harvesting date (68.1%), which
followed by the first date (57.7%) and both the second and fourth ones
(52.8 and 52.1%, respectively).

Data illustrated in Table (15) demonstrated the acceptance


percentages of grafted larvae (%) in each bar during harvesting dates
for Carniolian hybrid queenless production colonies grafted with
different genetic origin larvae. In each bar, significant or highly
significant differences were found between percentages of

91
Table 14. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in harvesting dates for
Carniolian hybrid queenless production colonies grafted with different genetic origin larvae,
throughout the period extended from 6/8/2008 to 15/8/2008.
production frame

Genetic origin of
Harvesting date

grafted larvae

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/

Harvested royal jelly amount


Accepted grafted larvae (%)
(gm) / race (hybrid)/ harvesting
/race (hybrid)/harvesting date
date

1st
3 Carniolian race 48 57.7 b 1.121 a
6/8/2008
92

2nd
3 Carniolian race 48 52.8 c 1.098 a
9/8/2008
3rd
3 Carniolian hybrid 48 68.1 a 0.669 b
12/8/2008
4th
3 Carniolian race 48 52.1 c 0.391 c
15/8/2008
General mean 57.7 0.820
*
F value 5.20 33.14 **
LSD 0.05 4.2 0.203
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

92
70 1.4
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


Harvested royal jelly
60 1.2
larvae
larvae %

50 1
grafted
grafted

40 0.8
of accepted
Meanof%accepted

93
30 0.6

20 0.4
Mean

10 0.2

0 0
06/08/2008 09/08/2008 12/08/2008 15/08/2008
Harvesting dates

Fig. 17. Fig.(%)


Mean 17. of
Means of accepted
accepted graftedgrafted
larvae larvae percentages
and harvested andjelly
royal harvested royal
amounts jellyinamounts
(gm) (gm)dates
harvesting in for
Carniolian hybrid queenless production colonies grafted with different genetic origin larvae throughout
harvesting dates for Carniolian hybrid queenless production colonies grafted with differegenetic the
period fromlarvae
origin 6-8-2008 to 15-8-2008.
throughout the period from 22-3-2008 to 31-3-2008
93
acceptance larvae throughout the four harvesting dates.

The sequences of acceptance percentages were the same in both


the first and second bars. However, the highest percentages occurred in
the third harvesting date (66.7 and 83.4%, respectively). The fourth
date ranked the second order (45.9 and 77.1%, respectively). The
second date came in the third level (39.6 and 66.7%, respectively). The
lowest acceptance took place in the first harvesting date in which the
respective percentages were 31.3 and 54.2%.

Considering the third bar, it could be observed that the greatest


percentage happened in the first harvesting date (87.5%), followed by
both the third and second dates in which 54.2 and 52.1%, respectively
were recorded. The fourth date came in the third level having 33.4%.
The estimated mean percentage of acceptance / colony was 57.7%.

b. Amount of harvested royal jelly


According to the results of harvested royal jelly amounts presented in
Table (13), the variations among 3 bars of each production frame were
significant or highly significant. On the other hand, it could be noticed
that the trend of harvested royal jelly amounts in the three bars of each
production frame were nearly similar to those observed in cases of
accepted grafted larvae percentages except that was occurred in the
fourth production frame.

In the first production frame, the highest amount of harvested


royal jelly was gained in the third bar (1.396 gm), while the lowest
amount of harvested royal jelly (0.745 gm) was recorded in the first
bar. The second one had an intermediate rank reaching 1.222 gm.

94
Table 15. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in each bar during
harvesting dates for Carniolian hybrid queenless production colonies grafted with different genetic
origin larvae, throughout the period extended from 6/8/2008 to 15/8/2008.
production frame

Genetic origin of
Harvesting date

Accepted grafted larvae (%) /race Harvested royal jelly amount (gm)/
grafted larvae

No. of grafted
(hybrid) /bar
larvae / race
No. of bars/

(hybrid)/bar/harvesting date race (hybrid)/ bar/harvesting date

No. of bar No. of bar

1st 2nd 3rd 1st 2nd 3rd

1st
3 Carniolian race 48 31.3 d 54.2 d 87.5 a 0.745 a 1.222 b 1.396 a

95
6/8/2008
2nd
3 Carniolian race 48 39.6 c 66.7 c 52.1 b 0.671 b 1.444 a 1.179 b
9/8/2008
3rd Carniolian
3 48 66.7 a 83.4 a 54.2 b 0.515 c 1.073 c 0.419 d
12/8/2008 hybrid
4th
3 Carniolian race 48 45.9 b 77.1 b 33.4 c 0.497 c 0.133 d 0.544 c
15/8/2008
General mean 45.9 70.4 56.8 0.607 0.968 0.885
** * ** ** **
F value 11.77 5.38 19.51 7.62 24.79 15.12**
LSD 0.05 6.1 9.8 3.6 0.038 0.129 0.113
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

95
During the second production frame, the maximum amount of
harvested royal jelly occurred in the second bar (1.444 gm), followed
with by the third one (1.179 gm), whereas the first bar represented the
minimum amount (0.671 gm).
The bars in the third production frame could be arranged
descendingly according to their amount of harvested royal jelly as
follows; the 2nd, 1st and 3rd bars by having 1.073, 0.515 gm and 0.419
gm, respectively.

The highest amount of harvested royal jelly for the fourth


production frame bars was gained in the third bar (0.544 gm), followed
by the first one (0.497 gm) and finally the second one (0.133 gm).

Obtained results illustrated in Table (14) and Fig. (17), revealed


high significant variation among harvested royal jelly amounts
throughout the harvesting dates. In this respect, the highest royal jelly
amounts were gained in both the first and second harvesting dates
(1.121 and 1.098 gm, respectively) followed by the third and fourth
dates (0.669 and 0.391 gm, respectively).
Highly significant values were statistically proved when
concerning the harvested royal jelly amounts throughout harvesting
dates for each bar (Table 15).

In the first and third bars, the highest amounts of royal jelly
were got in the first harvesting date (0.745 and 1.396 gm, respectively).
The second date came in the second category, where the respective
values were 0.671 and 1.179 gm. The third category represented in the
third and fourth harvesting dates for the first bar (0.515 and 0.497 gm,

96
respectively), as for the third bar it represented in the fourth harvesting
date (0.544 gm), while the fourth rank achieved by the third one (0.419
gm).

As for the second bar, more amount of royal jelly (gm) was
gained in the second date (1.444 gm), followed descendingly by the
first, third and fourth dates in which 1.222, 1.073 and 0.133 gm royal
jelly, respectively were obtained. In general, the mean amount of
harvested royal jelly / colony was 0.820 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
The results of statistical analysis shown in Table (16) demonstrate
simple correlations (r) and simple regressions (b) of effect of accepted
grafted larvae percentage on amount of harvested royal jelly and effect
of laying workers percentage and number of combs covered with bees
(C.S.) on both acceptance percentage and amount of harvested
royal jelly for Carniolian hybrid queenless production colonies that
grafted with different genetic origin larvae, throughout the period
extended from 6/8/2008 to 15/8/2008.
Those data showed that the influence of mean percentages of
grafted larvae on mean amounts of royal jelly (gm) was insignificant.
As for the effect of laying workers, results of "r' and "b" values
indicated that this factor has an insignificant effect in case of accepted
grafted larvae but it has significantly negative influence in case of
harvested royal jelly ("t" = 4.647). .As regards of the calculated “t”
values of the effects of number of combs covered with bees on both

97
Table 16. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Carniolian hybrid queenless production colonies
which grafted with different genetic origin larvae, throughout the period extended from 6/8/2008 to
15/8/2008. Harvested royal jelly amount

Accepted Laying workers Number of combs covered with bees


Accepted grafted larvae

grafted larvae
Accepted grafted Harvested royal Accepted grafted Harvested royal
on harvested
Harvesting date

larvae jelly larvae jelly


royal jelly

Mean percentage
(gm)
(%)

Mean number
98

r b t r b t r b t r b t r b t

1st
57.7 1.121 30 7
6/8/2008
2nd
52.8 1.098 50 7.2

4.647 *
0.096ns

0.040ns

0.308ns
-0.068

-0.003

-0.957

-0.013

-0.213

-0.011

-0.954

-1.042

4.483*
-0.03

-0.01

9/8/2008
3rd
68.1 0.669 70 7.4
12/8/2008
th
4
52.1 0.391 90 7.9
15/8/2008
ns
Non significant
* Significant at probability level of 0.05
98
accepted grafted larvae and harvested royal jelly, it could be noticed
that the simple correlation and regression values were insignificant in
the former case but they were negative and significant in the later case
("t" = 4.483).
Generally, it could be seen from results of Carniolian hybrid
queenless production colonies which grafted with different genetic
origin larvae that there were significant and highly significant
differences in the mean acceptance (%) and mean amount (gm) of RJ /
colony among the different grafted bars represented the tested race and
hybrid of bees at the same harvesting date.
From the data concerning the three bars, it could be observed
that the first bar had the minimum percentage of acceptance and
amount of RJ. The second one gave the highest percentage and amount.
The third bar recorded an intermediate percentage of acceptance and
the amount of RJ.
The means of acceptance percentage and amount of harvested
royal jelly for those colonies were 57.7% and 0.820 gm, respectively.

The effect of accepted grafted larvae percentage on the amount


of harvested royal jelly was insignificant, while the effects of each of
laying workers percentage and number of combs covered with bees
(C.S.) on acceptance percentage were insignificant while their effects
on the amount of harvested royal jelly were significant.

99
5. Egyptian race grafted with different genetic origin larvae
throughout the period extended from 22/3/2009 to 6/4/2009
a. Acceptance percentage of grafted larvae
Results of statistical analysis for the means of accepted grafted
larvae % in bars for Egyptian race queenless production colonies
grafted with different genetic origin larvae (Table, 17) showed
significant variations among the 3 bars of the 1st and 3rd production
frames, while the differences among bars were highly significant in
cases of the 4th, 5th and 6th production frames. Conversely, no
significant differences were obtained relating the 3 bars of the 2nd
production frame.

In the first production frame, comparatively high acceptance


percentages were recorded in both the second (Carniolian race) and
first (Italian hybrid) bars (33.3 and 31.3%, respectively) as compared
with that occurred in the third (Egyptian race) bar (2.1%).

In the third production frame, the accepted grafted larvae


percentages reached its maximum in the second bar (47.9%), followed
by the first bar in which 33.4% was happened then the third one
(27.1%).
The percentage of the fourth production frame was higher in the
first bar (68.8%) when compared with those of both the second and
third ones which recorded (29.2% and 27.1%, respectively.
Results shown in Table (17) also revealed that the utmost
percentage of acceptance for the 5th production frame was gained in the
first bar (70.9%), while the least percentage occurred in the third bar

100
Table 17. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in bars for Egyptian race
queenless production colonies grafted with different genetic origin larvae throughout the period
extended from 22/3/2009 to 6/4/2009.

Genetic origin

(hybrid) /bar
larvae / race
No. of bars/
production
Harvesting

of grafted
Accepted grafted larvae (%) Harvested royal jelly amount (gm)

grafted
larvae

No. of
frame
date

/race (hybrid)/bar /race(hybrid)/bar

Mean F value LSD 0.05 Mean F value LSD 0.05


Italian hybrid 16 31.3 a 0.090 b
1st *
3 Carniolian race 16 33.3 a 6.59 5.5 0.160 a 6.77 * 0.052
22/3/2009
Egyptian race 16 2.1 b 0.007 c
Carniolian race 16 10.4 0.073 a
2nd
0.38 ns 5.99 *

101
3 Egyptian race 16 6.3 0.047 b 0.022
25/3/2009
Italian hybrid 16 8.3 0.024 c
33.4 b 0.455
3rd *
3 Egyptian race 48 47.9 a 8.74 4.9 0.646 0.66 ns
28/3/2009
27.1 c 0.380
68.8 a 1.192 a
4th
3 Carniolian race 48 29.2 b 75.68 ** 4.2 0.582 b 18.08 **
0.027
31/3/2009
27.1 b 0.294 c
70.9 a 1.359 a
5th
3 Italian hybrid 48 58.3 b 57.81 ** 8.6 1.182 b 40.34 **
0.084
3/4/2009
6.3 c 0.064 c
41.7 c 0.762 b
6th
3 Italian hybrid 48 77.1 a 36.20 ** 6.4 1.292 a 19.70 ** 0.032
6/4/2009
58.3 b 0.725 c
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

101
(6.3%), whereas the second bar gave significantly an intermediate
value (58.3%).

As for the sixth production frame, the maximum percentage of


acceptance was achieved in the second bar (77.1%), followed by the
third one (58.3%) and finally the first bar (41.7%).

Results illustrated in Table (18) and Fig. (18) revealed that the
harvesting dates had high significant effects on the acceptance of
grafted larvae percentages for Egyptian race queenless production
colonies grafted with different genetic origin larvae. Also, it was
observed that the mean percentages of larvae acceptance increased
significantly with the progress of harvesting from the third to the sixth
dates.
However, the highest mean percentage of 59.0% occurred in the last
harvesting date. The fifth date ranked the second order with 45.2% on
average. In the fourth, third and first dates, the average acceptance
were 41.7, 36.1 and 22.2% representing the 3rd , 4th and 5th ranks,
respectively. The minimum acceptance was recorded in the second
harvesting date (8.4%).
The mean percentages of accepted grafted larvae in each bar
throughout the six harvesting dates for Egyptian race queenless
production colonies grafted with different genetic origin larvae are
demonstrated in Table (19). Analysis of variance indicated that
differences between acceptance percentages for each bar in the
different harvesting dates were highly significant, where the resulting
"F" values calculated 22.91, 20.75 and 75.97 in the three bars,

102
Table 18. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in harvesting dates for
Egyptian race queenless production colonies grafted with different genetic origin larvae, throughout
the period extended from 22/3/2009 to 6/4/2009.

larvae / race
No. of bars/
production
Harvesting

origin of

(hybrid)
Genetic
grafted

grafted
Accepted grafted larvae Harvested royal jelly amount

larvae

No. of
frame
date

/bar
(%) /race (gm)/ race (hybrid)/ harvesting
(hybrid)/harvesting date date

Italian hybrid 16
1st
3 Carniolian race 16 22.2 e 0.086 e
22/3/2009
Egyptian race 16
Carniolian race 16
2nd

103
3 Egyptian race 16 8.4 f 0.048 f
25/3/2009
Italian hybrid 16
3rd
3 Egyptian race 48 36.1 d 0.494 d
28/3/2009
4th
3 Carniolian race 48 41.7 c 0.690 c
31/3/2009
5th
3 Italian hybrid 48 45.2 b 0.868 b
3/4/2009
6th
3 Italian hybrid 48 59.0 a 0.926 a
6/4/2009
General mean 35.4 0.519
F value 35.82 ** 36.25 **
LSD 0.05 3.4 0.035
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

103
60 1.8
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


Harvested royal jelly
50 1.5
larvae
grafted %
larvae

40 1.2
grafted
of accepted

30 0.9
%accepted
104

20 0.6
Meanof
Mean

10 0.3

0 0
22/03/2009 25/03/2009 28/03/2009 31/03/2009 03/04/2009 06/04/2009
Harvesting dates

Fig. 18. MeanFig.


(%)18.
of accepted
Means ofgrafted larvae
accepted and harvested
grafted royal jellyand
larvae percentages amounts (gm) royal
harvested in harvesting dates for
jelly amounts Egyptian
(gm)
race in
queenless production colonies grafted with different genetic origin larvae throughout the
harvesting dates for Egyptian race queenless production colonies grafted with different period from
22- 3-2009
genetictoorigin
6-4-2009.
larvae throughout the period from 22- 3-2009 to 6-4-2009.
104
respectively. It could be stated that the acceptance percentages changed
according to genetic origin used for grafting as shown in the same table
which elucidate the following;

In the first bar, the highest acceptance was gained in both the
fifth and fourth harvesting dates (70.8 and 68.8%, respectively)
followed by the sixth date which recorded 41.7%. Both the third and
first dates came in the third level (33.4% and 31.3%, respectively. The
lowest acceptance was gained in the second date (10.4%).

Considering the second bar, the sixth harvesting date gave the
highest acceptance (77.1%), followed downwardly with the fifth, third,
first, fourth and second dates which recorded 58.3, 47.9, 33.3, 29.2 and
6.3%, respectively.

As for the third bar, the resulting acceptance percentage values


clarified that the maximum percentage of acceptance was gained at the
sixth harvesting date (58.3%), followed by both the third and fourth
ones (27.1 and 27.1%, respectively). The second and fifth dates had the
third rank, where 8.3 and 6.3%, respectively were attained. The first
date had the last level (2.1%). As for the mean percentage of
acceptance / colony it recorded 35.4%.

b. Amount of harvested royal jelly


Results of harvested royal jelly amounts (gm) in bars for
Egyptian race queenless production colonies grafted with different
genetic origin larvae that illustrated in Table (17), demonstrated that
there were significant or highly significant differences among the three
bars of harvesting dates excluded that calculated in the 3rd harvesting

105
Table 19. Mean (%) of accepted grafted larvae and harvested royal jelly (gm) amounts in each bar during
harvesting dates for Egyptian race queenless production colonies grafted with different genetic origin
larvae, throughout the period extended from 22/3/2009 to 6/4/2009.

grafted larvae

No. of grafted
(hybrid) /bar
Harvested royal jelly amount

larvae / race
No. of bars/
production
Harvesting

Accepted grafted larvae (%) /race

origin of
Genetic
(gm) / race (hybrid)/

frame
(hybrid)/bar/harvesting date
date

bar/harvesting date
No. of bar No. of bar
1st 2nd 3rd 1st 2nd 3rd
Italian hybrid 16
1st
3 Carniolian race 16 31.3 c 33.3 d 2.1 d 0.090 e 0.160 d 0.007 d
22/3/2009
Egyptian race 16
106

Carniolian race 16
2nd
25/3/2009
3 Egyptian race 16 10.4 d 6.3 f 8.3 c 0.073 e 0.047 e 0.024 d
Italian hybrid 16
3rd
28/3/2009 3 Egyptian race 48 33.4 c 47.9 c 27.1 b 0.455 d 0.646 c 0.380 b
4th
31/3/2009 3 Carniolian race 48 68.8 a 29.2 e 27.1 b 1.192 b 0.582 c 0.294 c
5th
3/4/2009 3 Italian hybrid 48 70.8 a 58.3 b 6.3 c 1.359 a 1.182 b 0.064 d
6th
6/4/2009 3 Italian hybrid 48 41.7 b 77.1 a 58.3 a 0.762 c 1.292 a 0.725 a
General mean 42.7 42.01 21.5 0.655 0.651 0.249
** ** ** ** ** **
F value 22.91 20.75 75.97 36.83 44.41 5.48
LSD 0.05 4.3 3.7 4.3 0.035 0.092 0.079
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

106
date in which variations among 3 bars were insignificant.
Obtained results showed similar tendency of royal jelly yielding
in the three bars of both the first and sixth production frames, where the
royal jelly was collected in relatively high amounts in the 2nd bar (0.160
and 1.292 gm, respectively. A decline in royal jelly gathering was
recorded in the first bar (0.090 and 0.762 gm, respectively). The lowest
harvested royal jelly was attained in the third bar (0.007 and 0.725 gm,
respectively).
Also, the harvesting royal jelly had the same trend in the three
bars of the 2nd, 4th and 5th harvesting dates. Collect on of royal jelly in
the first bar was the highest as compared with those gathered in both
the second and third bars, Thus, the highest amounts were got in the
first bar (0.073, 1.192 and 1.359 gm, respectively) followed by those
obtained in the second bar (0.047, 0.582 and 1.182 gm, respectively).
The third bar ranked the third group, where the corresponding royal
jelly amounts were 0.0 24, 0.294 and 0.064 gm.

With regarding "F" value for harvested royal jelly amounts


shown in Table (18), the results proved highly significant variations
among mean harvested royal jelly amounts throughout the six
harvesting dates. The same table also clarified that collected royal jelly
had a similar trend to that occurred in the case of accepted grafted
larvae percentages. Significantly, the highest amount of royal jelly
(0.926 gm) was gathered in the sixth date. This date was followed in
descending orders as follows; the 5th, 4th, 3rd, 1st and 2nd dates in which
0.868, 0.690, 0.494, 0.086 and 0.048 gm, respectively of royal jelly
were gathered.

107
Considering royal jelly amount averages in each bar (Table 19),
highly significant differences were calculated among harvested royal
jelly in the six harvesting dates for each bar.

In case of the first bar, the highest amount of harvested royal


jelly was gained in the fifth date (1.359 gm), followed descendingly
with the fourth, sixth and third harvesting dates that recorded 1.192,
0.762 and 0.455 gm, respectively. Both the first and second dates came
in the last order, where 0.090 and 0.073 gm of royal jelly were gained,
respectively).

As for the second bar, the sixth harvesting date represented the
first rank, where the maximum amount (1.292 gm) were collected. The
fifth date came in the second rank (1.182 gm). Both the third and
fourth dates had the third rank where 0.646 and 0.582 gm were gained,
respectively. The fourth rank was represented by the first harvesting
date (0.160 gm), that followed by the second date (0.047 gm).

In the third bar, the harvested royal jelly could be arranged


downward as follows; the sixth date gave the highest mean amount of
royal jelly (0.725 gm). This date was followed descendingly by the
third harvesting date with 0.380 gm, the fourth date which gave 0.294
gm. The fifth, second and first harvesting dates came in the last group
where comparatively low amounts of royal jelly were collected (0.064,
0.024 and 0.007 gm, respectively). On the other hand, the mean amount
of harvested royal jelly / colony was 0.519 gm.

108
c. Effect of accepted grafted larvae percentage on amount of
harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
Table (20) gives simple correlations (r) and simple regressions
(b) of effect of accepted grafted larvae percentage on amount of
harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly for Egyptian race
queenless production colonies which grafted with different genetic
origin larvae, throughout the period extended from 22/3/2009 to
6/4/2009.

The data in this table show that the (r) and (b) values for the
effect of the accepted grafted larvae percentages on harvested royal
jelly amounts (gm) were highly significant.

As regards of the calculated “t” values for the effect of laying


workers, the results indicated that this factor had highly significant and
positive effects on both accepted grafted larvae percentages and harvested
royal jelly amounts ("t" values were 5.385 and 7.077, respectively).

When considering effect of number of combs covered with bees,


the resulting "t" values denoted that the influence of the mentioned
factor was highly significant in the case of accepted grafted larvae
percentages and harvested royal jelly.

It noticeably to summarize the previous results for Egyptian race


queenless production colonies grafted with different genetic origin
larvae as significant or highly significant differences were recorded for

109
Table 20. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Egyptian race queenless production colonies
grafted with different genetic origin larvae, throughout the period extended from 22/3/2009 to 6/4/2009.
Accepted grafted Laying workers Number of combs covered with bees

Harvested royal jelly


Accepted grafted
larvae on harvested Accepted grafted Harvested royal Accepted grafted Harvested royal
Harvesting dates

amount (gm)
royal jelly
larvae (%)
larvae jelly larvae jelly

percentage

number
Mean

Mean
r b t r b t r b t r b t r b t
110

1st 22.2 0.086 50 4


22/3/2009
nd
2 8.4 0.048 50 4.2
25/3/2009
rd
3
6.420**

5.385**

7.077**

4.796**

8.833**
36.1 0.494 55 4.6
0.955

0.020

0.937
2.052

0.962

0.045

0.923
0.030

0.975

1.480
28/3/2009
th
4 41.7 0.69 60 4.9
31/3/2009
th
5 45.2 0.868 65 5.2
3/4/2009
th
6 59.0 0.926 70 5.5
6/4/2009
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01

110
the mean acceptance percentage and mean amount (gm) of RJ / colony
among the different grafted bars represented the tested race and hybrid
of bees at the same harvesting date.
As for the acceptance percentage and the amount of harvested RJ,
the highest one occurred for the Italian hybrid followed successively
with the Carniolian race then the Egyptian one.
It could be seen from results of the three bars that the maximum
percentage of acceptance and amount of RJ occurred in the first bar
followed by the second one, whereas the lowest percentage of
acceptance and the amount of RJ happened in the third bar.
The mean percentage of acceptance and mean amount of
harvested royal jelly for those colonies were 35.4% and 0.519 gm,
respectively.

The effect of accepted grafted larvae percentage on the amount


of harvested royal jelly was highly significant. On the other hand, the
effects of each of laying workers percentage and number of combs
covered with bees (C.S.) on both acceptance percentage and amount of
harvested RJ were highly significant.

Generally, the following points could be concluded from


previous results concerning the colonies grafted with non related
larvae;

The results of the three bars of grafting frame, showed that the
highest mean of acceptance percentage occurred in the second bar
(55.2%), while the lowest mean percentage detected in the third one
(42.8%). The first bar had an intermediate mean percentage of 50.4%.

111
As for the results of amount of harvested royal jelly, the greatest mean
amount was harvested from the first bar followed successively by the
second then the third ones (0.841, 0.836 and 0.701%, respectively).
These results are in harmony with the findings of Sharaf El-Din
et al. (2000) in Egypt. They detected that when grafted queen cups
were installed on the lower level of bars, the highest percentages of
acceptance of grafted larvae were induced. The least results were
obtained when grafted queen cups were fixed on the upper bar. The
differences between effects of the three levels (lower, middle or upper)
of bars were mostly significant.
On the other hand, different results had been demonstrated by
Visscher (1986) who studied the kinship discrimination using larvae for
queen rearing. In his experiments, he transferred larvae into royal jelly
in queen cups, and had relatively high acceptance (57%) and no
evidence of nepotism was found. The bees reared nearly equal number
of unrelated larvae (77) and related nestmate larvae (76). These
differences in the results could be referred to the differences in the
environmental conditions and genetic origin of the reared bees.

The general mean of acceptance percentage for the above


colonies was 49.6%, whereas the general mean amount of harvested
royal jelly was 0.804%.

The majority of tested colonies indicated insignificant influence


of accepted grafted larvae percentage on amount of harvested royal
jelly. In addition, the effects of laying workers percentage on number
of combs covered with bees (C.S.) on both acceptance percentage and

112
amount of harvested royal jelly were insignificant except for the fifth
colony (Egyptian race which carried out during the period extended
from 22/3/2009 to 6/4/2009) as it showed highly significant relations.
The highly significant influence could be happened in this colony due
to the excess number of harvesting dates that may be resulted in high
percentage of laying workers and affected the means of acceptance
percentage and amount of harvested royal jelly.
These results are not in agreement with those reported by Rana
et al. (1996) in India who recorded that the yield of royal jelly is related
to the weight of royal jelly. Also, Durmus and Guler (1999) in Turkey,
who found that colony size has significant effects on royal jelly yield.
As a final point, it could be stated that the sharp reduction of
area cultivated with plants and the ecological factors have partially
effects on the production colonies as well as the genetic origin of
production colonies may be affect the tested colonies.

Furthermore, the appearance of laying workers of the Egyptian


race colonies is often happened prior to other colonies, thus the degree
of developed ovaries is always senior on the degrees of the other
colonies (El-Enany, 2006).

b. Related grafted larvae


Those experiments carried out utilizing Egyptian race, Italian
and Carniolian hybrids queenless production colonies. Each race or
hybrid was used as donor and production colony. Three bars were used
in each production frame. After evaluating larvae acceptance % and
royal jelly production (gm), the used bar was replaced by another one.

113
1. Egyptian race used as donor and production colonies
throughout the period extended from 8/4/2009 to 20/4/2009
a. Acceptance percentage of grafted larvae
Results in Table (21) presented means of accepted grafted larvae
percentages and harvested royal jelly amounts (gm) in bars for
Egyptian race production colonies.
According to the calculated "F" values, there were significant or
highly significant differences among the three bars of the 1st, 2nd, 3rd,
and 4th production frames. Reversely, there wasn‟t significant
difference among the 3 bars of the 5th production frame.
In the case of the first production frame, the incidence of
maximum larvae acceptance percentage took place in the 3rd bar
(77.1%) followed by the 2nd one then the 1st bar (41.7 and 12.5%,
respectively).
The trend of acceptance larvae percentages in the second
production frame were similar to that happened in the first one. The
respective percentages of larvae acceptance were 77.1, 60.4 and 29.2%.
The occurrence of highest percentage of larvae acceptance in the
third production frame was achieved in both 3rd and 2nd bars where 22.9
and 18.8%, respectively were obtained . Comparatively low percentage
was took place in the 1st bar (4.2%).
Relatively, maximum larvae acceptance percentage of the fourth
production frame occurred in the 3rd bar (54.2%) followed by both 1st
and 2nd bars that gave the same acceptance percentage (35.4%).
Statistical analysis results shown in Table (22) and

114
Table 21. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in bars for Egyptian race
queenless production colonies used as donor and production colonies throughout the period extended
from 8/4/2009 to 20/4/2009.
Harvesting date

Genetic origin

No. of grafted
No. of bars/
production
Accepted grafted larvae (%) Harvested royal jelly amount (gm)

of grafted

/race/bar
larvae
larvae
frame
/race/bar /race/bar

Mean F value LSD 0.05 Mean F value LSD 0.05

1st 12.5 c 0.481 c


3 Egyptian race 48 41.7 b 35.79 ** 5.25 1.507 b 29.73 **
0.095
8/4/2009
77.1 a 2.763 a

115
2nd 29.2 c 0.880 c
3 Egyptian race 48 60.4 b 22.73 ** 7.17 1.969 b 16.30 **
0.083
11/4/2009
77.1 a 2.512 a
3rd 4.2 b 0.015 c
3 Egyptian race 48 18.8 a 8.41 * 5.78 0.636 b 25.05 **
1.060
14/4/2009
22.9 a 0.875 a
4th 35.4 b 0.806 b
3 Egyptian race 48 35.4 b 5.42 * 5.80 0.617 c 31.70 **
1.082
17/4/2009
54.2 a 1.189 a
5th 4.2 0.152
3 Egyptian race 48 6.3 0.27 ns 0.141 0.02 ns
20/4/2009
8.4 0.130
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).
115
graphically illustrated in Fig. (19) revealed that harvesting dates high
significant affected means of accepted larvae percentages of Egyptian
race. The accepted grafted larvae percentages ranged between 6.3 %
and 55.6 % according to harvesting dates.
According to which are shown in Table (22) and Fig (19) it
could be stated that the five harvesting dates could be arranged
descendingly according to their acceptance percentages as follows: the
second harvesting date gave the highest acceptance (55.6%) as
compared with acceptance percentages obtained in other dates. Both
the first and fourth dates came in the second group with 43.8 and
41.7%, respectively. Then the third date came after with 15.3%
followed by the fifth date that represented the last rank with 6.3%
acceptance.
Consideration of results of statistical analysis listed in Table
(23), it may be interest to point out that there were highly significant
variations among of accepted grafted larvae percentages in each bar
during the five harvesting dates.

In the first bar, the maximum percentage of acceptance was


recorded in the fourth harvesting date (35.4%), followed downwardly
by the second and first dates which recorded 29.2 and 12.5%,
respectively. The third and fifth dates representing the same group and
came in the last level (4.2 and 4.2%, respectively).

With minor differences in acceptance percentage values, the


occurrence of accepted grafted larvae in the 3rd, 4th and 5th harvesting
dates in the second and third bars had the same trend.

116
Table 22. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in harvesting dates for
Egyptian race queenless production colonies used as donor and production colonies throughout the
period extended from 8/4/2009 to 20/4/2009.

larvae /race
No. of bars/
production
Harvesting

origin of
Genetic
grafted

grafted
larvae

No. of
frame
date

Accepted grafted larvae (%) Harvested royal jelly amount (gm)

/bar
/race/harvesting date /race/ harvesting date

1st
3 Egyptian race 48 43.8 b 1.583 b
8/4/2009
2nd
3 Egyptian race 48 55.6 a 1.787 a
11/4/2009

117
3rd
3 Egyptian race 48 15.3 c 0.509 d
14/4/2009
4th
3 Egyptian race 48 41.7 b 0.871 c
17/4/2009
5th
3 Egyptian race 48 6.3 d 0.141 e
20/4/2009
General mean 32.5 0.978
**
F value 49.21 46.96 **
LSD 0.05 4.5 0.201
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

117
60 1.8
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


Harvested royal jelly
larvae
50 1.5
larvae
grafted %

40 1.2
grafted
of accepted

30 0.9
accepted
118

Meanof%

20 0.6
Mean

10 0.3

0 0
08/04/2009 11/04/2009 14/04/2009 17/04/2009 20/04/2009
Harvesting dates

Fig. 19. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in harvesting dates for
Fig. 19. Means of accepted grafted larvae percentages and harvested royal jelly amounts (gm)
Egyptian race queenless production colonies used as donor and production colonies throughout the
in harvesting dates for Egyptian hybrid queenless production colonies used as donor and
period from 8-4-2009 to 20-4-2009.
production colonies throughout the period from 8-4-2009 to 20-4-2009.

118
Table 23. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Egyptian race queenless production colonies used as donor and production colonies
throughout the period extended from 8/4/2009 to 20/4/2009.
Harvesting dates

Genetic origin of

larvae /race /bar


grafted larvae

No. of grafted
No. of bars/
production Accepted grafted larvae (%) Harvested royal jelly amount (gm)
frame /race/bar/harvesting date /race/ bar/harvesting date

No. of bar No. of bar


st
1 2nd 3rd
1st
2nd 3rd
1st
3 Egyptian race 48 12.5 c 41.7 b 77.1 a 0.481 c 1.507 b 2.763 a
8/4/2009

119
2nd
3 Egyptian race 48 29.2 b 60.4 a 77.1 a 0.880 a 1.969 a 2.512 a
11/4/2009
3rd
3 Egyptian race 48 4.2 d 18.8 c 22.9 c 0.015 e 0.636 c 0.875 c
14/4/2009
4th
3 Egyptian race 48 35.4 a 35.4 b 54.2 b 0.806 b 0.617 c 1.189 b
17/4/2009
5th
3 Egyptian race 48 4.2 d 6.3 d 8.4 d 0.152 d 0.141 d 0.130 d
20/4/2009
General mean 17.1 32.5 47.9 0.466 0.974 1.494
F value 15.10 ** 23.91 ** 43.65 ** 48.305 ** 21.065 ** 228.51 **
LSD 0.05 5.3 6.7 7.0 0.065 0.094 0.258
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different(LSD test).

119
In case of the second bar, the maximum percentage occurred in
the second date where 60.4% of acceptance was recorded followed
with the first date which had 41.7% of acceptance. Considering the
third bar, it was noticed that the first and second dates had the same
values of acceptance (77.1%) and considered as the first group in which
highest acceptance percentages were recorded .
The corresponding acceptance means for the 4th, 3rd and 5th
harvesting dates in both the second and third bars were 35.4 and 54.2%,
18.8 and 22.9%, as well as 6.3 and 8.4%, respictively. In another point
of view mean percentage of acceptance / colony was 32.5%.

b. Amount of harvested royal jelly


The results shown in Table (21) on the 3 bars for Egyptian race
queenless production colonies, showed that there were highly
significant differences between mean amounts of gathered royal jelly
in the three bars of the first four harvesting dates. On the other hand,
the calculated "F" value was insignificant in the fifth date.
In this regard, it was noticed that the occurrence of collected
royal jelly was similar in the 3 bars of the first, second and third dates.
The 3rd bar showed high amounts of harvest with 2.763, 2.512 and
0.875 gm in the previous three dates, respectively. The 2nd bar came in
the second rank, whereas the respective amounts recorded 1.507, 1.969
and 0.636 gm. The lowest amounts were gained in the 1st bar in which
0.481, 0.880 and 0.015 gm of royal jelly were attained, respectively.
In the fourth production frame (date), the bars could be arranged
descendingly as the 3rd bar which got the highest amount of harvested

120
royal jelly (1.189 gm) followed by the 1st one then the 2nd bar (0.806
and 0.617 gm, respectively).
Results presented in Table (22) and illustrated graphically in
Fig. (19) revealed the occurrence of highly significant variations among
collected royal jelly throughout the five studied harvesting dates. These
dates could be arrange as follows:
The second date gave the maximum mean amount of royal jelly
of 1.787 gm. The first date represented the second rank with 1.583 gm.
The fourth, third and fifth dates came in the 3rd, 4th and 5th ranks, where
0.871, 0.509 and 0.141gm of royal jelly were gathered, respectively.
Data illustrated in Table (23) further indicated that the amounts
of harvested royal jelly (gm) in each bar throughout harvesting dates
for Egyptian race queenless production colonies were highly
significant.
The highest amount of collected royal jelly (gm) in the first bar
was gained at the second harvesting date (0.880 gm), followed in
descending orders with the fourth, first, fifth and third dates which
recorded 0.806, 0.481, 0.152 and 0.015 gm, respectively.
In the second bar, the maximum royal jelly amount was
recorded in the second date (1.969 gm), followed by the first one which
recorded 1.507 gm . Both the third and fourth ones denoted the third
rank (0.636 and 0.617 gm, respectively). The last level was the fifth
harvesting date with 0.141 gm.
In case of the third bar, both the first and second dates gave the
highest amounts of royal jelly (2.763 and 2.512 gm, respectively). The
fourth, third and fifth harvesting dates represented the 2nd, 3rd and 5th

121
ranks in which 1.189, 0.875 and 0.130 gm of royal jelly were collected,
respectively. Meanwhile the mean amount of harvested royal jelly /
colony was 0.978 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
Results in Table (24) representing the simple correlations (r) and
simple regressions (b) of effect of accepted grafted larvae percentage
on amount of harvested royal jelly and effect of laying workers
percentage and number of combs covered with bees (C.S.) on both
acceptance percentage and amount of harvested royal jelly for
Carniolian hybrid queenless production colonies which grafted with
different genetic origin larvae, throughout the period extended from
6/8/2008 to 15/8/2008.
Obtained data indicated that primary influence of accepted
grafted larvae % on harvested royal jelly amount throughout the five
harvesting dates was positive and significant ("t" value was 4.722).
On the other hand, it was interesting to note that the primary
effects of laying workers were insignificant in both cases of accepted
grafted larvae and harvested royal jelly amounts.
As for the influence of number of combs covered with bees, the
resulting "t" values indicated that this factor also had insignificant
effects on both cases of accepted grafted larvae and harvested royal
jelly amounts.
It could be summarize the above results as the different grafted
bars were significantly or high significantly affected the mean

122
Table 24. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Egyptian race queenless production colonies used
as donor and production colonies, throughout the period extended from 8/4/2009 to 20/4/2009.

Harvested royal jelly amount


Accepted Laying workers Number of combs covered with bees
Accepted grafted larvae
grafted larvae
Accepted grafted Harvested royal Accepted grafted Harvested royal
on harvested
Harvesting date

larvae jelly larvae jelly


royal jelly

Mean percentage
(gm)
(%)

Mean number
r b t r b t r b t r b t r b t

123
1st 43.8 1.583 60 4
8/4/2009
nd
2
55.6 1.787 80 4.2
11/4/200

1.110 ns

1.985 ns

1.559 ns

2.732 ns
-0.540

-0.720

-0.754

-0.034

-0.669

-0.013

-0.845

-0.480
4.722*

rd
0.939

0.032

39
15.3 0.509 85 4.4
14/4/200
th
49
41.7 0.871 95 4.7
17/4/200
59th
6.3 0.141 100 5
20/4/200
ns
9Insignificant
* Significant at probability level of 0.05

123
acceptance (%) and the mean amount (gm) of RJ / colony in the same
harvesting date.
It is apparent from the results of the three bars that the first bar
gave the smallest percentage of acceptance and amount of RJ, reversely
to the third one which recorded the highest percentage and amount.
The second bar had an intermediate percentage of acceptance the
amount of harvested RJ.
Results considering the general means of acceptance percentage
and amount of harvested royal jelly for these colonies were 32.5% and
0.978 gm, respectively.

The effect of accepted grafted larvae percentage on harvested


royal jelly amount was significant. On the reverse, the effects of each
laying workers percentage and number of combs covered with bees
(C.S.) on both acceptance percentage and amount of harvested royal
jelly were insignificant.

2. Italian hybrid used as donor and production colonies


throughout the period extended from 15/7/2009 to 2/8/2009
a. Acceptance percentage of grafted larvae
Results in Table (25) present means of accepted grafted larvae
% and harvested royal jelly amounts (gm) in bars for Egyptian race
production colonies.

According to those data, significant or highly significant


differences were detected between means of accepted grafted larvae %
throughout the harvesting dates except for the second date which
denoted non-significant variations.

124
In the first production frame (harvesting date), great acceptance
grafted larvae percentage took place in the 3rd bar (85.4%) then the first
one (58.4% ) and thirdly the second bar (35.4%).

In the third production frame, the highest percentage of


acceptance was gained in the 1st bar (64.6%), followed with both the
second and third ones which recorded same percentages (41.7 and
41.7%, respectively).

In the three bars of the fourth and seventh production frames, the
incidences of acceptance percentages were coincident. In this regard,
considerably large acceptance percentages took place in the 2nd bar
(95.8 and 85.4%, respectively). The second rank of acceptance
happened in the 3rd bar in which the corresponding percentages were
85.5 and 58.3%. The 1st bar represented the last order, where 72.9 and
41.7% of acceptance occurred.

In addition, there was a synchronization in the acceptance


occurrences in the three bars of the fifth and sixth production frames.
The maximum percentage was recorded in the 3rd bar with 89.6% in the
fifth frame and 83.3% in the sixth one. This percentage was followed
by those recorded in the 2nd bar (56.3 and 77.1%, respectively). In the
1st bar, acceptance got its minimum percentages (8.3 and 58.3%,
respectively).

Results for Italian hybrid queenless production colonies used as


production and donor colonies showed highly significant differences
among acceptance percentages of grafted larvae in harvesting dates
(Table 26 and Fig. 20).

125
Table 25. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in bars for Italian hybrid
queenless production colonies used as donor and production colonies throughout the period extended
from 15/7/2009 to 2/8/2009.

No. of bars/
production
Harvesting

hybrid/bar
origin of
Harvested royal jelly amount

Genetic

larvae /
grafted

grafted
larvae
frame

No. of
Accepted grafted larvae (%) /hybrid/bar
date

(gm))/hybrid/bar

Mean F value LSD 0.05 Mean F value LSD 0.05


58.4 b 1.393 b
1st
3 Italian hybrid 48 35.4 c 46.67 ** 8.6 0.925 c 32.87 **
0.442
15/7/2009
85.4 a 2.063 a
66.7 0.689 c
2nd
3 Italian hybrid 48 70.9 1.88 ns 1.036 b 12.83 **
0.054
18/7/2009
79.2 1.099 a
64.6 a 1.086 a
3rd
5.75 * **
126

3 Italian hybrid 48 41.7 b 8.5 0.856 b 21.65 0.219


21/7/2009
41.7 b 0.617 c
72.9 c 1.433 b
4th
3 Italian hybrid 48 95.8 a 10.12 ** 6.9 1.481 a 22.05 **
0.047
24/7/2009
85.5 b 0.760 c
8.3 c 0.071 c 0.227
5th
3 Italian hybrid 48 56.3 b 30.49 ** 9.2 0.956 b 41.26 **
27/7/2009
89.6 a 1.230 a
58.3 c 0.653 c
6th
3 Italian hybrid 48 77.1 b 19.61 ** 5.4 1.227 a 44.98 **
0.102
30/7/2009
83.3 a 1.121 b
41.7 c 0.532
7th
3 Italian hybrid 48 85.4 a 16.01 ** 8.5 0.474 1.34 ns
2/8/2009
58.3 b 0.586
ns
Non significant
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different(LSD test).

126
The fourth harvesting date came in the first order (84.7%). Both
the sixth and second dates ranked the second group (72.9 and 72.2%,
respectively) followed by the third order which represented by both the
seventh and first harvesting dates in which 61.8 and 59.7% were
recorded, respectively. Finally, the fifth and third dates represented the
same group in which the corresponding percentages were 51.4% and
49.3% respectively.

The results of analysis of variance for means of accepted grafted


larvae percentages in each bar during the seven harvesting dates are
shown in Table (27). This table referred that the variations among
acceptance percentages in each bar - as measured by F test - were
highly significant in all studied harvesting dates.
In the first bar, the maximum of acceptance percentage took
plane in the 4th harvesting date (72.9%). The second group represented
with the 2nd and 3rd dates, which recorded 66.7 and 64.6%, respectively.
The third group occurred in both the first and sixth ones, where they
had the same acceptance value (58.4 and 58.4%, respectively). The
fourth rank occurred in the 7th date with 41.7%.The last order of 8.4%
acceptance occurred in the 5th date.
Regarding the second bar, considerably highest percentage of
acceptance (95.8%) was determined in the 4th harvesting date with.
This percentage was followed in descending orders by the 7th, 6th, 2nd
and 5th dates in which 85.4, 77.1, 70.9 and 56.3% of acceptance were
recorded, respectively. The 3rd and 1st dates had the least means of
acceptance (41.7 and 35.6%, respectively).

127
Table 26. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in harvesting dates for
Italian hybrid queenless production colonies used as donor and production colonies throughout the
period extended from 15/7/2009 to 2/8/2009.

No. of bars/

/hybrid/bar
Harvesting

production

origin of
Genetic
grafted

grafted
Accepted grafted larvae

larvae
larvae

No. of
frame
date

Harvested royal jelly amount


(%)/hybrid/harvesting
(gm) /hybrid/ harvesting date
date

1st 3 Italian hybrid 48 59.7 c 1.460 a


15/7/2009
2nd 3 Italian hybrid 48 72.2 b 0.941 d
128

18/7/2009
3rd 3 Italian hybrid 48 49.3 d 0.852 e
21/7/2009
4th 3 Italian hybrid 48 84.7 a 1.225 b
24/7/2009
5th 3 Italian hybrid 48 51.4 d 0.752 f
27/7/2009
6th 3 Italian hybrid 48 72.9 b 1.000 c
30/7/2009
7th 3 Italian hybrid 48 61.8 c 0.531 g
2/8/2009
General mean 64.6 0.959
**
F value 32.71 42.82 **
LSD 0.05 5.3 0.052
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

128
90 2.7
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


80 Harvested royal jelly 2.4
%
larvae
larvae

70 2.1
grafted

60 1.8
grafted
of accepted

50 1.5
accepted

40 1.2

129
of%

30 0.9
Mean
Mean

20 0.6

10 0.3

0 0
15/07/2009 18/07/2009 21/07/2009 24/07/2009 27/07/2009 30/07/2009 02/08/2009
Harvesting dates

Fig. 20. Mean (%)


Fig. 20.of Means
accepted
of grafted
accepted larvae andlarvae
grafted harvested royal jelly
percentages andamounts (gm)
harvested in harvesting
royal dates(gm)
jelly amounts for Italian
hybrid queenless dates
in harvesting production colonies
for Italian usedqueenless
hybrid was donor and production
production colonies
colonies throughout
used was the period from
donor and
15- 7-2009 to 2-8-2009.
production colonies throughout the period from 15- 7-2009 to 2-8-2009.
129
Table 27. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Italian hybrid queenless production colonies used as donor and production
colonies, throughout the period extended from 15/7/2009 to 2/8/2009.

No. of grafted
No. of bars/

/hybrid/bar
Accepted grafted larvae (%) Harvested royal jelly amount (gm)
production
Harvesting

origin of
Genetic
grafted
larvae

larvae
frame
/hybrid/bar/harvesting date /hybrid/ bar/harvesting date
date

No. of bar No. of bar


st
1 2nd 3rd
1st
2nd 3rd
1st 3 Italian hybrid 48 58.4 c 35.6 f 85.5 b 1.393 a 0.925 d 2.063 a
15/7/2009
2nd
3 Italian hybrid 48 66.7 b 70.9 d 79.2 c 0.689 c 1.036 c 1.099 c
130

18/7/2009
3rd
3 Italian hybrid 48 64.6 b 41.7 f 41.7 e 1.086 b 0.856 e 0.617 e
21/7/2009
4th
3 Italian hybrid 48 72.9 a 95.8 a 85.5 b 1.433 a 1.481 a 0.760 d
24/7/2009
5th
3 Italian hybrid 48 8.4 e 56.3 e 89.6 a 0.071 e 0.956 d 1.230 b
27/7/2009
6th
3 Italian hybrid 48 58.4 c 77.1 c 83.4 b 0.653 c 1.227 b 1.121 c
30/7/2009
7th
3 Italian hybrid 48 41.7 d 85.4 b 58.4 d 0.532 d 0.474 f 0.586 f
2/8/2009
General mean 53.0 66.1 74.7 0.743 0.993 1.068
** ** ** **
F value 21.54 26.94 18.31 48.23 44.14 ** 36.37 **
LSD 0.05 4.0 4.7 4.1 0.103 0.062 0.034
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

130
The highly acceptance mean of the third bar occurred in the fifth
harvesting date (89.6%). The 1st, 4th and 5th dates represented the same
group with 85.5, 85.5 and 83.4% acceptance, respectively. The second
harvesting date ranked in the third order (79.2%). The fourth level was
reserved for the 7th harvesting date as it recorded 58.4%. The last level
took place in the 3rd date with 41.7%. For overall view, the mean
percentage of acceptance / colony was 64.6%.

b. Amount of harvested royal jelly


Data of harvesting royal jelly amounts in bars for Egyptian race
production colonies presented in Table (25) denoted that there were
highly significant variations between amounts means (gm) in the three
bars of the earliest six harvesting dates (production frames). On the
reverse, no significant differences were calculated among the three bars
of the 7th date.
The highest amount of gathered royal jelly (2.063 gm) was
gained in the 3rd bar of the first harvesting date followed by the 1st and
2nd bars, respectively in which the particular percentages were 1.393
and 0.925 gm.
The trend of harvested royal jelly (gm) gathered in the three bars
of the second and fifth frames (dates) were nearly in accordance. Those
percentages can be arranged in downward levels as the 3rd, 2nd and 1st
bars, respectively. In those bars, the corresponding values of royal jelly
amounts recorded 1.099 and 1.230 gm, 1.036 and 0.956 gm and 1.036
and 0.071 gm.
As for the third production frame, the royal jelly gained its
maximum in the 1st bar, where it averaged 1.086 gm followed with the

131
2nd bar with 0.856 gm then the 3rd bar which came in the last rank with
0.617 gm.
In the fourth and sixth production frames, the three bars can be
arranged in descending orders according to their harvested royal jelly
as follows; the 2nd bar had greatest amounts (1.481 and 1.227 gm,
respectively) as compared with those gathered from the other two bars.
The second level represented by the 1st in case of fourth frame (1.443
gm) or the 3rd bar (1.121 gm) or that of the sixth frame. The third order
distinguished by the 3rd bar or 1st one in case of the fourth or sixth
frame in which 0.760 gm or 0.653 gm of royal jelly was gained,
respectively.
An examination of results concerning harvested royal jelly
amounts in harvesting dates illustrated in Table (26) and Fig. (20), the
statistical analysis denoted highly significant between collected royal
jelly amounts in harvesting dates.
The seven cardinal directions could be arranged descendingly
according to their harvested amounts of royal jelly as follows;
The 1st harvesting date attained the highest mean amount of
royal jelly (1.460 gm). This date was followed descendingly by the 4th
one which gave 1.225 gm, the 6th date with 1.000 gm, the 2nd date with
0.941 gm, the 3rd one with 0.852 gm, the 5th date with 0.752 gm. The
least amount was gained in the 7th date in which 0.531 gm of royal jelly
was collected.
As shown in obtained results in Table (27), it can be stated that
the harvesting dates were high significantly affected the amounts of
harvested royal jelly in each bar.

132
In the first bar, the highest amounts of royal jelly were gained in
the 4th and 1st harvesting dates (1.433 and 1.393 gm, respectively), The
3rd date came in the next level with 1.086 gm. The third rank was
reserved for both the 2nd and 6th dates with 0.689 and 0.653 gm,
respectively, followed by the 7th one (0.532 gm) and finally the 5th
harvesting date with 0.071 gm.

Results of "F" value for the second bar indicated that relatively
high quantity of royal jelly was gathered in the 4th harvesting date
(1.481 gm). In the remaining harvesting dates, the 6th date came in the
next level with 1.227 gm, followed by the 2nd date which recorded
1.036 gm. The fourth group was reserved for both the 5th and 1st dates
that got 0.956 and 0.925 gm of royal jelly, respectively. The 3rd date
came in the fifth level (0.856 gm) followed by the 7th date in which
0.474 gm was recorded.

Regarding the third bar, it is worth noticing that maximum royal


jelly was collected at the first harvesting date (2.063 gm), followed by
the 5th date that recorded 1.230 gm. Both the 6th and 2nd dates came in
the third rank with 1.121 and 1.099 gm, respectively. The fourth rank
was hold for the 4th date with 0.760 gm, followed downwardly by the
3rd and 7th dates that recorded 0.617 and 0.586 gm, respectively. On the
contrary, the mean amount of harvested royal jelly / colony was 0.959
gm.

133
c. Effect of accepted grafted larvae percentage on amount of
harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
Results of statistical analysis of "t' values in Table (28) revealed
the simple correlations (r) and simple regressions (b) of effect of
accepted grafted larvae percentage on amount of harvested royal jelly
and effect of laying workers percentage and number of combs covered
with bees (C.S.) on both acceptance percentage and amount of
harvested royal jelly for Italian hybrid queenless production colonies
used as donor and production colonies throughout the seven harvesting
dates.

The simple correlations and regressions coefficients were


insignificant - when the effect of grafted larvae percentages on amounts
of harvested royal jelly - was studied (Table 28).

The same table further indicated that the effect of laying workers
throughout different dates was insignificant on both cases of accepted
grafted larvae and harvesting royal jelly amounts.

Moreover, the resulting "t" values for the influence of number of


combs covered with bees on acceptance percentages were also
insignificant in both cases of accepted grafted larvae and harvesting
royal jelly amounts.

Previous results revealed that there were significant and highly


significant differences for the mean acceptance (%) and mean amount
(gm) of RJ / colony among the different grafted bars represented the
tested hybrid of bees in the same harvesting date.

134
Table 28. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Italian hybrid queenless production colonies used
as donor and production colonies, throughout the period extended from 15/7/2009 to 5/8/2009.
Accepted grafted Laying workers Number of combs covered with bees

Harvested royal jelly


Accepted grafted
larvae on harvested Accepted grafted Harvested royal Accepted grafted Harvested royal
Harvesting date

amount (gm)
royal jelly
larvae (%)
larvae jelly larvae jelly

Mean number
percentage
Mean
r b t r b t r b t r b t r b t

1st

135
59.7 1.460 60 3
15/7/2009
2nd
72.2 0.941 60 3.3
18/7/2009
rd
3
49.3 0.852 70 3.5
21/7/2009
0.800ns

0.115ns

1.909ns

0.122ns

2.333ns
-0.649

-0.013

-0.722

-1.305
4th
0.337

0.008

0.051
0.043

0.054

0.002
84.7 1.225 80 3.7
27/7/2009
5th
51.4 0.752 85 4
30/7/2009
6th
72.9 1.000 90 4.3
2/8/2009
th
7
61.8 0.531 100 4.5
5/8/2009
ns
Non significant

135
As for the three bars, it could be noticed that the chief
percentage of acceptance and amount of RJ occurred in the third bar,
whereas the first bar had the lowest percentage and amount. The second
bar recorded middle values of acceptance percentage and amount of
harvested RJ.
Results for Italian hybrid queenless production colonies used as
production and donor colonies showed highly significant differences
among acceptance percentages of grafted larvae in harvesting dates.
The mean percentage of acceptance for this colony was 64.6%
while the mean amount of harvested royal jelly was 0.959 gm.

The influence of accepted grafted larvae percentage on amount


of harvested royal jelly was insignificant. Considering the effect of
each laying workers percentage and number of combs covered with
bees (C.S.) on both acceptance percentage and amount of harvested
royal jelly, obtained results were also insignificant.

3. Carniolian hybrid used as donor and production colonies


throughout the period extended from 15/7/2009 to 2/8/2009
a. Acceptance percentage of grafted larvae
This study was conducted throughout the period extended from
13-8-2008 until 28-8-2008. The means of accepted grafted larvae
percentages and harvested royal jelly amounts (gm) in bars that
calculated each 3 days are listed in Table (29).
According to the results of "F values" for acceptance
percentages throughout the six harvesting dates, it is evident that the

136
differences among three bars were highly significant in the 2nd, 3rd, 4th
5th and 6th production frames.
In the 1st one, no significant variations were determined among
its 3 bars.
In the second production frame, greatest percentage of
acceptance was gained in the 2nd bar (79.2%), followed by the 3rd then
the 1st ones with 73.0 and 47.9%, respectively.
The acceptance percentages in the third frame, pointed out that
the utmost of acceptance occurred in the 3rd bar (89.6%) that
descending with the 2nd bar (85.4%) and subsequent the 1st one
(45.8%).
As for means of accepted grafted larvae percentages that gained
in the fourth production frame, it has been found that both the 2nd and
3rd bars represented the first group with maximum percentages of
77.1and 73.0%, respectively. The 1st bar came in the second rank in
which 45.9% of acceptance was recorded.
In case of the fifth production frame, the highest percentages of
acceptance were 79.2 and 72.9% took place in the 3rd and 1st bars,
respectively, followed by the 2nd bar with 45.8% of acceptance.
The three bars of the sixth production could be arranged
downwardly according to their acceptance percentages as the 1st, 2nd
and 3rd bars in which 85.4, 54.2 and 39.6% were gained, respectively.
When variations between means of accepted grafted larvae
percentages and harvested royal jelly amounts (gm) in the different
harvesting dates studied, data shown in Table (30) and illustrated
graphically in Fig. (21) clarified highly significant differences.

137
Table 29. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in bars for Carniolian
hybrid queenless production colonies used as donor and production colonies throughout the period
extended from 13/8/2009 to 28/8/2009.

Genetic origin of
Harvesting date

larvae / hybrid
grafted larvae

No. of grafted
No. of bars/
production
Accepted grafted larvae Harvested royal jelly amount

frame
(%)/hybrid/bar (gm)/hybrid/bar

/bar
Mean F value LSD 0.05 Mean F value LSD 0.05

1st 89.6 2.513 a


3 Carniolian hybrid 48 89.6 0.13 ns 2.089 c 14.63 ** 0.105
13/8/2009 91.7 2.341 b
2nd 47.9 c 1.029 c
138

3 Carniolian hybrid 48 79.2 a 12.57 ** 5.8 1.646 a 24.17 ** 0.235


16/8/2009 73.0 b 1.394 b
3rd 45.8 c 0.926 c
3 Carniolian hybrid 48 85.4 b 22.41 ** 4.1 2.260 a 26.69 **
0.104
19/8/2009 89.6 a 2.069 b
4th 45.9 b 1.052 c
3 Carniolian hybrid 48 77.1 a 22.18 ** 4.9 2.034 a 17.16 **
0.142
22/8/2009 73.0 a 1.218 b
5th 72.9 a 1.617 a
3 Carniolian hybrid 48 45.8 b 12.08 ** 6.4 1.118 b 11.81 **
0.071
25/8/2009 79.2 a 1.133 b
6th 85.4 a 1.909 a
3 Carniolian hybrid 48 54.2 b 25.24 ** 7.8 0.808 b 29.45 **
0.204
28/8/2009 39.6 c 0.473 c
ns
Non significant
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

138
The acceptance percentages varied in the different harvesting
dates. The maximum acceptance mean took place in the 1st date
(89.8%) followed by the 3rd date in which 73.6% of acceptance was
attained. The three harvesting dates of the 2nd, 5th and 4th ones
represented the same group that came in the third rank with 66.7, 66.0
and 65.3%, respectively. The 6th date recorded the least acceptance
percentage (59.7%).
Results tabulated in Table (31) showed means of acceptance
percentages and amounts of collected royal jelly (gm) in each bar
during harvesting dates. The statistical analysis (F. values) showed
highly significant difference between means of accepted grafted larvae
in each bar during the six harvesting dates.
In each bar, the harvesting dates could be arranged in an
ascending order according to its acceptance percentage means as
follows:
In case of the first bar, The 1st group contained the 1st and 6th
dates with 89.4 and 85.4 %, respectively followed by the 5th date which
recorded 72.9%. The 2nd, 3rd and 4th dates represented the same level
and reserved for the third group where 47.9, 45.9 and 45.9% of
acceptance were occurred, respectively.
As for the second bar, both the 1st and 3rd dates came in the first
group where the maximum acceptance took place (89.4 and 85.4%,
respectively) followed by the 2nd and 4th dates together with
corresponding acceptances of 79.2 and 77.1%, then the 6th date
(54.2%). In the 5th harvesting date, relatively low acceptance was
attained (45,8%).

139
Table 30. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in harvesting dates for
Carniolian hybrid queenless production colonies used as donor and production colonies throughout
the period extended from 13/8/2009 to 28/8/2009.

No. of grafted
/hybrid /bar
No. of bars/
Harvesting

production

origin of
Genetic
grafted

larvae
larvae
frame
dates

Accepted grafted larvae Harvested royal jelly amount


(%)/hybrid/harvesting date (gm) /hybrid/ harvesting date

1st 3 Carniolian hybrid 48 89.8 a 2.314 a


13/8/2009
2nd 3 Carniolian hybrid 48 66.7 c 1.356 ce
16/8/2009
140

3rd 3 Carniolian hybrid 48 73.6 b 1.751 b


19/8/2009
4th 3 Carniolian hybrid 48 65.3 c 1.434 c
22/8/2009
5th 3 Carniolian hybrid 48 66.0 c 1.289 de
25/8/2009
6th 3 Carniolian hybrid 48 59.7 d 1.063 f
28/8/2009
General mean 70.2 1.535
F value 13.22 ** 11.77 *
LSD 0.05 2.6 0.088
* Significant at probability level of 0.05
** Highly significant at probability level of 0.01
Means within a column followed by different letter are significant or highly significant different (LSD test).

140
100 3
Accepted grafted larvae

Mean of harvested royal jelly amounts (gm)


90 2.7
Harvested royal jelly
Mean of accepted grafted larvae %
Mean % of accepted grafted larvae

80 2.4

70 2.1

60 1.8

50 1.5

40 1.2

141
30 0.9

20 0.6

10 0.3

0 0
13/08/2009 16/08/2009 19/08/2009 22/08/2009 25/08/2009 28/08/2009
Harvesting dates
Fig. 21. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in harvesting dates for
Fig. 21.hybrid
Carniolian Meansqueenless
of accepted grafted larvae
production percentages
colonies used was and harvested
donor royal jellycolonies
and production amountsthroughout
(gm) the
in harvesting
period datestofor28-8-2009.
from 13- 8-2009 Carniolian hybrid queenless production colonies used was donor and
production colonies throughout the period from 13- 8-2009 to 28-8-2009.
141
Table 31. Mean (%) of accepted grafted larvae and harvested royal jelly amounts (gm) in each bar during
harvesting dates for Carniolian hybrid queenless production colonies used as donor and production
colonies, throughout the period extended from 13/8/2009 to 28/8/2009.

/hybrid /bar
No. of bars/
production
Harvesting

Accepted grafted larvae (%) Harvested royal jelly amount (gm)

origin of
Genetic
grafted

grafted
larvae

larvae
No. of
frame
/hybrid/bar/harvesting date /hybrid/ bar/harvesting date
date

No. of bar No. of bar


st
1 2nd 3rd
1st
2nd 3rd
1st
3 Carniolian hybrid 48 89.4 a 89.4 a 91.9 a 2.513 a 2.089 b 2.341 a
13/8/2009
2nd
3 Carniolian hybrid 48 47.9 c 79.2 bc 73.0 b 1.029 d 1.646 d 1.394 c
142

16/8/2009
3rd
3 Carniolian hybrid 48 45.9 c 85.4 a 89.4 a 0.926 e 2.260 a 2.069 b
19/8/2009
4th
3 Carniolian hybrid 48 45.9 c 77.1 c 73.0 b 1.052 d 2.034 c 1.218 d
22/8/2009
5th
3 Carniolian hybrid 48 72.9 b 45.8 e 79.2 b 1.617 c 1.118 e 1.133 e
25/8/2009
6th
3 Carniolian hybrid 48 85.4 a 54.2 d 39.6 c 1.909 b 0.808 f 0.473 f
28/8/2009
General mean 64.5 71.9 74.4 1.507 1.659 1.438
F value 21.52 ** 16.06 ** 13.63 ** 29.15 ** 23.66 ** 15.89 **
LSD 0.05 4.86 5.55 5.94 0.102 0.045 0.077
** Highly significant at probability level of 0.01.
Means within a column followed by different letter are significant or highly significant different (LSD test).

142
It is observed from the results of the third bar that the 1st and 3rd
dates gave the most acceptances of grafted larvae (91.9 and 89.4%,
respectively) followed by the 5th, 2nd and 3rd harvesting date, where
the consequent acceptance recorded 79.2, 73.0 and 73.0% then the 6th
date that recorded the lowest acceptance of 39.6%.
According to previously presented data it could be stated that
mean percentage of acceptance / colony was 70.2%.

b. Amount of harvested royal jelly


From the results presented in Table (29) concerning the amounts
of harvested royal jelly, it appeared that the three bars of each
harvesting date (production frame) for Carniolian hybrid queenless
colonies varied with high significantly level in their collected amounts
of royal jelly.
In the first production frame, harvested royal jelly amount
reached its maximum in the 1st bar (2.513 gm), whereas the 3rd and 2nd
bars ranked the second and third orders with 2.341 and 2.089 gm,
respectively.
In cases of the second, third and fourth production frames, the
trend of royal jelly amounts in the three bars were in accordance. In this
regard, the 2nd bar gave the highest amount of royal jelly where the
related means recorded 1.646, 2.260 and 2.034 gm of royal jelly in the
three frames, respectively. This bar was followed with the 3rd one in
which gave 1.394, 2.096 and 1.218 gm, respectively. Relatively low
amounts of collected royal jelly were obtained in the 1st bar (1.029,
0.926 and 1.052 gm, respectively).
The maximum amount of harvested royal jelly with 1.617 gm

143
took place in the 1st bar of the fifth frame, followed by those collected
in the 3rd and 2nd bars together (1.133 and 1.118 gm, respectively).
As for the sixth production frame, it was observed that the
highest amount of harvested royal jelly was gained in the 1st bar (1.909
gm) as compared with that attained in the 2nd one (0.808 gm) that
followed by the 3rd bar (0.473 gm).
Harvested mean amounts of royal jelly were calculated each
three days throughout the period extended from August 13 to August
28 of 2008. Results of collected royal jelly in the six harvesting dates
are listed in Table (30) and illustrated in Fig (21). Significant
differences were found between the mean amounts of gathered royal
jelly throughout the six harvesting dates.
The 1st harvesting date gave the largest amount of royal jelly
where 2.314 gm was collected. The 3rd date came in the second level
where 1.751 gm was gained followed by the 4th date (1.434 gm). The
2nd and 5th dates were in the fourth order (1.356 and 1.289 gm,
respectively). In the 6th date, the lowest amount of royal jelly (1.063
gm) was attained.
Results of royal jelly amounts that gained in each bar during the
six harvesting dates are illustrated in Table (31). The resultant "F"
values proved highly significant variations between the collected royal
jelly in harvesting dates for each bar.
In the first bar, the highest amount of royal jelly (gm) was
gained in the 1st harvesting date (2.513), followed by the 6th then 5th
dates which recorded 1.909 and 1.617 gm, respectively. The fourth
rank was kept for both the 4th and 2nd dates (1.052 and 1.029 gm,

144
respectively). The 3rd harvesting date gave the lowest royal jelly
amount (0.926 gm).
In case of the second bar, harvested royal jelly attained its
maximum amount in the 3rd harvesting date (2.260 gm) that followed in
descending orders with the 1st, 4th, 2nd, 5th and 6th dates in which 2.089,
2.034, 1.646, 1.118 and 0.808 gm, of royal jelly were gained,
respectively.

The most amount of royal jelly in the third bar took place in the
1st date (2.341 gm), followed with the 3rd one then the 2nd, 4th, 5th and
6th dates with , 2.069, 1.394, 1.218, 1.133 and 0.473 gm, respectively.

Among the data showed previously, the mean amount of


harvested royal jelly / colony reached 1.535 gm.

c. Effect of accepted grafted larvae percentage on amount of


harvested royal jelly and effect of laying workers percentage and
number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly
The simple correlations (r) and regressions (b) of effect of
accepted grafted larvae percentage on amount of harvested royal jelly
and effect of laying workers percentage and number of combs covered
with bees (C.S.) on both acceptance percentage and amount of
harvested royal jelly for Carniolian hybrid queenless production
colonies used as donor and production colonies, throughout the period
extended from 13/8/2009 to 28/8/2009.

The simple correlation and regression values of mean percentages


of accepted grafted larvae on mean amounts of harvested royal jelly

145
(Table 32) were positive, where 't" value was highly significant
(12.397).

The simple correlation and regression values of mean percentages


of laying workers on mean percentages of accepted grafted larvae were
negative, where 't" value was insignificant (1.791), While it was
negative in the case of amount of harvested royal jelly, where 't" value
was insignificant (1.824).

The simple correlation and regression values of mean numbers of


combs covered with bees on mean percentages of accepted grafted
larvae were negative, where 't" value was insignificant (2.421). At the
same time, it was negative for mean amounts of harvested royal jelly,
where 't" value was insignificant (2.602).

According to results of Carniolian hybrid queenless production


colonies used as donor and production colonies, it could be detected
that there were highly significant differences between the mean
acceptance (%) and mean amount (gm) of RJ / colony in the different
grafted bars represented the tested hybrid of bees at the same
harvesting date
As for the three bars, it could be observed that the first bar gave
the minimum percentage of acceptance, whereas the third one had the
highest percentage of acceptance but the second bar recorded an in-
between percentage. As for amount of harvested RJ, the second bar had
the maximum value, followed with the first one while the third bar
came in the last rank.

146
Table 32. Simple correlations (r) and regressions (b) of effect of accepted grafted larvae (%) on harvested royal
jelly amount as well as effect of laying workers (%) and number of combs covered with bees on both
acceptance (%) and harvested royal jelly amount for Carniolian hybrid queenless production colonies
which used as donor and production colonies, throughout the period extended from 13/8/2009 to
28/8/2009.
Accepted grafted Laying workers Number of combs covered with bees
Accepted grafted larvae

larvae on harvested
Harvested royal jelly royal jelly Accepted grafted larvae
Harvested royal
Accepted grafted larvae Harvested royal jelly
Harvesting date

amount (gm) jelly

Mean percentage

Mean number
(%)

r b t r b t r b t r b t r b t

147
1st
89.8 2.314 60 3
13/8/2009
2nd
66.7 1.356 60 3.2
16/8/2009
3rd
12.397**

73.6 1.751 80 3.4

1.791 ns

1.824 ns

2.421 ns

2.602 ns
-0.667

-0.419

-0.674

-0.018

-0.771

-0.043

-0.793

-1.053
0.987

0.041

19/8/2009
4th
65.3 1.434 80 3.8
22/8/2009
5th
66 1.289 95 4.2
25/8/2009
6th
59.7 1.063 100 4.5
28/8/2009
ns
Non significant
** Highly significant at probability level of 0.01.

147
Regarding the mean percentage of acceptance and mean amount
of harvested royal jelly, they recorded 70.2% and 1.535 gm,
respectively.

Highly significant value was achieved for the effect of accepted


grafted larvae percentage on amount of harvested royal jelly. On the
other side, insignificant influences were calculated when studied the
effect of laying workers percentage and number of combs covered with
bees (C.S.) on acceptance percentage and amount of harvested royal
jelly which were insignificant.

Based on the above results concerning the colonies grafted with


related larvae, it could be generally stated the followings;
As for the three bars of grafting frame, the highest means of
acceptance percentage and amount of harvested royal jelly were gained
in the third bar (65.6% and 1.333 gm, respectively) while the lowest
mean values were detected in the first one (44.8% and 0.905 gm,
respectively). The second bar had an intermediate means 56.8% and
1.209 gm, respectively. These results are in accordance with the
findings of Sharaf El-Din et al. (2000) in Egypt, who reported that
when grafted queen cups were put on the lower level of bars, the
highest percentages of acceptance grafted larvae were recorded. The
least values were obtained when grafted queen cups were fixed on the
upper bar. The differences among effects of the three levels (lower,
middle or upper) of bars were mostly significant.

The general mean percentage of acceptance for these colonies


was 55.8%, while the mean amount of harvested royal jelly was
1.157%.

148
All tested colonies proved insignificant influences of accepted
grafted larvae percentage on amount of harvested royal jelly. As for the
effects of each laying workers percentage and number of combs
covered with bees (C.S.) on both acceptance percentage and amount of
harvested royal jelly, the resulted values were also insignificant. These
results are not agreed with those reported by Rana et al. (1996) in
India, who found that the yield of royal jelly was related to the weight
of royal jelly per cell and the number of queen cups in a colony. Also,
Durmus and Guler (1999) In Turkey who determined that colony size
had significant effects on royal jelly yield.
The general means and ranges of accepted grafted larvae
percentages and harvested royal jelly amounts (gm) for colonies grafted
with non related and related larvae were illustrated in Table (33) which
summarized and denoted the followings;
In case of accepted percentage for colonies grafted with non
related larvae it was observed that the highest mean percentage of
acceptance (57.7%) took place in the fourth colony (Carniolian hybrid)
which grafted with Carniolian race and Carniolian hybrid larvae. This
colony was followed by the second one (Carniolian hybrid) which
grafted with Carniolian race larvae (54.7%).
The first colony (Egyptian race) which grafted with different
genetic origin larvae (Italian hybrid, Carniolian and Egyptian races)
came in the third rank where it recorded 51.9% acceptance. The fourth
rank reserved for the third colony (Carniolian hybrid) which grafted
with Carniolian race and hybrid larvae where it had the mean
acceptance percentage of 48.1%. Finally, the Egyptian race (fifth

149
Table 33. General averages of accepted grafted larvae percentages and
harvested royal jelly amounts (gm) for colonies grafted with
non-related and related larvae.

larvae (%)

Harvested
royal jelly
Accepted

amount
grafted

(gm)
Genetic origin of Average Average
production colonies (Range) (Range)

Non Related grafted larvae


1- Egyptian race 51.9 0.438

2- Carniolian hybrid 54.7 1.103

3- Carniolian hybrid 48.1 49.6 1.141 0.804


(35.4 - 57.7) (0.438 - 1.141)
4- Carniolian hybrid 57.7 0.820

5- Egyptian race 35.4 0.519

Related grafted larvae


1- Egyptian race 32.5 0.978
55.8 1.157
2- Italian hybrid 64.6 0.959
(32.5 - 70.2) (0.959 - 1.535)
3- Carniolian hybrid 70.2 1.535

colony) which grafted with Italian hybrid, Carniolian and Egyptian race
came in the last order where it reached the least mean of acceptance
percentage (35.4%).
As for colonies grafted with related larvae that used as donor
and production ones, it was noticed that the highest mean of acceptance
percentage was detected in the Carniolian hybrid (third) colony
(70.2%) followed by the second (Italian hybrid) one in which 64.6%

150
was recorded. The first colony (Egyptian race) showed the least mean
percentage of acceptance (32.5%).
Regarding the amount of harvested royal jelly for colonies
grafted with non related larvae, it could be classified in a descending
order as follows; the third colony(Carniolian hybrid) had the chief
amount of 1.141 gm then the second one (Carniolian hybrid) came in
the second rank with 1.103 gm. The third grade represented by the
fourth colony (Carniolian hybrid) which gave 0.820 gm. The Egyptian
race (fifth colony) came in the fourth rank (0.519 gm). Finally, the first
colony (Egyptian race) reversed the last position (0.438 gm).
Colonies grafted with related larvae could be arranged
successively according to their amount of harvested royal jelly as
follows; the Carniolian hybrid colony ranked in the first order where it
had the highest amount of harvested royal jelly (1.535 gm), followed
by the Egyptian race in which 0.978 gm was gathered. The Italian
hybrid colony proved the least amount of harvested royal jelly (0.959
gm).
These results are in agreement with the findings of Laidlaw and
Eckert (1950) who noticed that the Italian races tend to produce fewer
queen cells than other races such as Carniolian. Also, KroI (1985) in
Poland who illustrated that during the first 6 days of queen larval
development, there were differences between Caucasian, Carniolian,
Italian and North European honey bee races, in addition Carniolians
built the largest and heaviest queen cells and put most royal jelly in
each cell. In all the races, the amount of royal jelly in the cell was
maximal on the fourth day of larval development.

151
In the same respect, Saleh (1999) showed that Craniolian bees
produced more royal jelly than the Italian bee race.
On the other hand, Albarracin et al. (2006) disagreed with this
results. They used two genetic groups (Africanized and Italian bees)
and three treatments of grafting T1 (larvae from the same genetic group
and the same colony), T2 (same genetic group and different colony),
and T3 (different genetic groups). They demonstrated that during the
process of queen production, the genetic groups and treatments showed
no significant differences in acceptance percentage of larvae and
weight of royal jelly for queen cells. Differences in results may be
attributed to the differences in the environmental conditions and
genetic origins of tested colonies.
As a general view of these results it could be said that the
colonies grafted with related larvae had a higher mean percentage of
acceptance (55.8%) which ranged from 32.5% to 70.2% than the
colonies grafted with non related larvae in which 49.6% and ranged
from 35.4% to 57.7%. From another point of view the mean amount of
harvested royal jelly was observed in the colonies grafted with related
larvae had a higher mean amount (1.157 gm) which ranged from 0.959
gm to 1.535 gm than the colonies grafted with non related larvae
reaching 0.804 gm ranging between 0.438 gm - 1.141 gm.
This means that the acceptance percentage and amount of
harvested royal jelly means was senior in the colonies grafted with
related larvae in the tested race and hybrids and similar to the results
stated by Page and Erickson (1984) who found that when workers
rearing queens were given a choice between which were their full

152
sister, and larvae from another colony to which they were related
approximately as closely as they would be to their half sisters, they
preferred to rear the more closely related larvae. However, this
discrimination may have been facilitated by differences in colony
odour between the two groups of larvae, while Mohammedi and Le
Conte (2000) showed that A. m. mellifera workers accepted
significantly more sisters (77.5%) than alien bees (20%), While A. m.
ligustica workers accepted 77.5% of sister workers and only 45% of A.
m. mellifera workers. This difference was significant confirming the
ability of the bees of the two races to discriminate.
Focusing the results obtained in this study, it concluded that
the amount of harvested royal jelly produced from honeybee colonies
of different race and hybrids of bees were generally low as compared
with previous studies that carried out in the apiary yard in Giza region
since 35 years. In this investigation, the mean amounts of harvested
royal jelly ranged between 0.048 and 2.472 gm / colony. While, in the
study of Eweis (1974), these amounts ranged between 2.990 - 14.230
gm / colony. Also, Abd Al-Fattah (1987) recorded 2.700 to 7.100 gm /
colony.
Minimum low amounts of royal jelly in this investigation may
be attributed to many reasons such as the sharp reduction of area
cultivated with plants; bees keeping that was mainly depended on the
artificial feeding. Also, the ecological factors have a partial effect on
the production colonies, as well as the genetic origin of production
colonies that may be affect clearly on the production of royal jelly.

153
In conclusion of this study, Carniolian hybrid colonies were
the favorable hybrid for royal jelly production, where it gave an
average of 1.141 gm royal jelly/colony as compared with Italian hybrid
and Egyptian race that gave 0.966 and 0.519 gm royal jelly/colony
through the same period.

2. Determination of amino and fatty acid percentages in samples of


the produced royal jelly
The analysis of royal jelly was carried out using samples taken
from Egyptian race, Carniolian and Italian hybrids queenless
production colonies. The purpose of this study was to determine
changes in amino and fatty acid percentages in the different race and
hybrids of honeybee. Each race or hybrid was used as production and
donor colonies (related colonies).
Harvested royal jelly samples from the production colonies were
stored in a deep freezer then analyzed to determine the type, and
percentages of different amino, and fatty acids in royal jelly.
a. Determination of amino acid percentages
In this study, the determined amino acids separated into two
main types; Essential amino acids and non-essential amino acids.
This study revealed the occurrence of sixteen amino acids (8
essential acids and 8 non-essential acids) in all tested samples. In
addition, analysis of royal jelly samples for essential and non essential
amino acids showed variations between the percentages of the same
amino acid in different samples at different harvesting numbers and
percentages of laying workers.

154
1. Egyptian race used as production and donor colonies.
a. Essential amino acids
Table (34) showed mean percentages of essential amino acids in
royal jelly produced from different harvesting numbers and percentages
of laying workers.
In the first harvesting date (1st sample) in which 60% of laying
workers happened, the essential amino acid means were as follows:
Threonine 0.571%, Valine 0.742%, Methionine 0.132%, Isoleucine
0.501%, Leucine 1.086%, Phenylalanine 0.406%, Histidine 0.375%
and Lysine 1.105%.
As for the 2nd sample, in which the laying workers percentage
was 80%, the essential amino acids (Threonine, Valine, Methionine,
Isoleucine, Leucine, Phenylalanine, Histidine and Lysine) mean
percentages reached 0.529, 0.672, 0.143, 0.470, 0.908, 0.342, 0.320 and
0.876% respectively.
The third sample was taken from the 3rd harvesting date when
the percentage of laying workers was 85%. The mean percentages of
Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine,
Histidine and Lysine acids were 0.500, 0.611, 0.061, 0.430, 0.862,
0.360, 0.349 and 0.971%, respectively.
According to essential amino acids in the fourth sample where
laying workers percentage was 95% , the afore-mentioned different
means of essential amino acid percentages reached 0.620, 0.830, 0.159,
0.584, 1.127, 0.456, 0.402 and 1.074%, respectively.

155
Table 34. Mean percentages of essential amino acids in royal jelly
samples produced from different harvesting numbers and
different percentages of laying workers for Egyptian race
colonies used as production and donor colonies.
Sample No. 1 2 3 4 5
Mean
Harvesting No. 1st 2nd 3rd 4th 5th
Percentage of laying
60 80 85 95 100 84
workers
Threonine 0.571 0.529 0.500 0.620 0.625 0.569
Type and percentage of essential

Valine 0.742 0.672 0.611 0.830 0.847 0.740

Methionine 0.132 0.143 0.061 0.159 0.085 0.116


amino acids

Isoleucine 0.501 0.470 0.430 0.584 0.551 0.507

Leucine 1.086 0.908 0.862 1.127 1.132 1.023

Phenylalanine 0.406 0.342 0.360 0.456 0.444 0.402

Histidine 0.375 0.320 0.349 0.402 0.463 0.382

Lysine 1.105 0.876 0.971 1.074 1.439 0.569

In case of the fifth sample the laying workers percentage was


100%, and the essential amino acids means recorded 625, 0.847, 0.085,
0.551, 1.132, 0.444, 0.463 and 1.439% for Threonine, Valine,
Methionine, Isoleucine, Leucine, Phenylalanine, Histidine and Lysine,
respectively.
When calculating the general means of essential amino acids, it
recorded 0.569% for Threonine, 0.740% for Valine, 0.116% for
Methionine, 0.507% for Isoleucine, 1.023% for Leucine, 0.402% for
Phenylalanine, 0.382% for Histidine and 0.569% for Lysine, while the
mean percentage of laying workers was 71%.

156
b. Non essential amino acids
The following points observed for mean percentages of non-
essential amino acids as shown in Table (35):
As for the first sample (1st harvesting date) where 60% of laying
workers took place, the non essential amino acids reached 2.582 ,
0.979, 1.144, 0.014, 0.920, 0.671, 0.448 and 0.490% for Aspartic,
Serine, Glutamic, Proline, Glycine, Alanine, Tyrosine and Arginine
acids, respictively.
In the 2nd sample when the laying workers percentage was 80%,
the respective means of non essential amino acids were 2.293, 0.930,
0.901, 0.012, 0.808, 0.624, 0.364 and 0.427%.
Considering the third sample that taken when the percentage of
laying workers was 85%, the mean percentages of Aspartic, Serine,
Glutamic, Proline, Glycine, Alanine, Tyrosine and Arginine acids were
2.240, 0.871, 1.032, 0.008, 0.808, 0.588, 0.380 and 0.408%,
respectively.
In the fourth sample with 95% of laying workers percentage,
2.733, 1.149, 1.289, 0.017, 1.007, 0.711, 0.501 and 0.512% were
attained for different non-essential amino acids successively.
At the fifth harvesting date in which the laying workers
percentage recorded 100%, the mean percentages of non essential
amino acids (Aspartic, Serine, Glutamic, Proline, Glycine, Alanine,
Tyrosine and Arginine) were 1.661, 1.106, 0.966, 0.022, 1.203, 0.784,
0.511 and 0.562%, respectively.

157
Table 35. Mean percentages of non-essential amino acids in royal jelly
produced from different harvesting numbers and different
percentages of laying workers by Egyptian race colonies when
used as production and donor colonies.

Sample No. 1 2 3 4 5
Mean
st nd rd th th
Harvesting No. 1 2 3 4 5
Percentage of laying
60 80 85 95 100 84
workers
Aspartic 2.582 2.293 2.240 2.733 1.661 2.302
Type and percentage of non essential

Serine 0.979 0.930 0.871 1.149 1.106 1.007

Glutamic 1.144 0.901 1.032 1.298 0.966 1.068


amino acids

Proline 0.014 0.012 0.008 0.017 0.022 0.015

Glycine 0.920 0.808 0.808 1.007 1.203 0.949

Alanine 0.671 0.624 0.588 0.711 0.784 0.676

Tyrosine 0.448 0.364 0.380 0.501 0.511 0.441

Arginine 0.490 0.427 0.408 0.512 0.562 0.480

The calculated general means of non essential amino acids were


as follows; Aspartic (2.230%), Serine (1.007%), Glutamic (1.068%),
Proline (0.015%), Glycine (0.949%), Alanine (0.676%), Tyrosine
(0.441%) and Arginine (0.480%). At the same time, the mean
percentage of laying workers was 84%.

158
2. Italian hybrid used as production and donor colonies.
a. Essential amino acids
The results of this study are demonstrated in Table (36), it
showed that mean percentages of essential amino acids varied
according to harvesting dates and laying workers percentages.
The mean percentages of Threonine, Valine, Methionine,
Isoleucine, Leucine, Phenylalanine, Histidine and Lysine acids in the
first sample when the percentage of laying workers recorded 60% were
0.562, 0.722, 0.127, 0.514, 1.000, 0.378, 0.337 and 0.844%,
respectively.
Results of the second sample with 80% laying workers recorded
0.541, 0.733, 0.116, 0.490, 0.956, 0.369, 0.346 and 0.801% for the
prior essential amino acids consecutively.
As for the seventh harvesting date (with 100% laying workers)
in which the third sample was taken, the mean percentages of
abovementioned eight essential amino acids recorded 0.424, 0.562,
0.091, 0.389, 0.746, 0.267, 0.255 and 0.644%, respectively.
In addition, Table (35) clarified that when laying workers
percentage averaged 80%, the means of essential amino acids were as
follows; 0.509, 0.672, 0.111, 0.464, 0.901, 0.338, 0.313 and 0.763% for
Aspartic, Serine, Glutamic, Proline, Glycine, Alanine, Tyrosine and
Arginine acids. respictivel.

159
Table 36. Mean percentages of essential amino acids in royal jelly
produced from different harvesting numbers and different
percentages of laying workers by Italian hybrid colonies when
used as production and donor colonies.

Sample No. 1 2 3
Mean
st rd th
Harvesting No. 1 3 7

Percentage of laying workers 60 80 100 80


Type and percentage of essential amino

Threonine 0.562 0.541 0.424 0.509

Valine 0.722 0.733 0.562 0.672

Methionine 0.127 0.116 0.091 0.111

Isoleucine 0.514 0.490 0.389 0.464


acids

Leucine 1.000 0.956 0.746 0.901

Phenylalanine 0.378 0.369 0.267 0.338

Histidine 0.337 0.346 0.255 0.313

Lysine 0.844 0.801 0.644 0.763

b. Non essential amino acids


Results concerning the mean percentages of non-essential amino
presented in Table (37) indicated that once percentage of laying
workers denoted 60% as presented in the first harvesting date, the non-
essential Aspartic, Serine, Glutamic, Proline, Glycine, Alanine,
Tyrosine and Arginine amino acids reached 2.516, 0.957, 0.971, 0.014,
0.901, 0.655, 0.394 and 0.468%, respectively.

160
Table 37. Mean percentages of non-essential amino acids in royal jelly
produced from different harvesting numbers and different
percentages of laying workers by Italian hybrid colonies when
used as production and donor colonies.

Sample No. 1 2 3
Mean
Harvesting No. 1st 3rd 7th

Percentage of laying workers 60 80 100 80

Aspartic 2.516 2.439 1.900 2.285


Type and percentage of non essential

Serine 0.957 1.005 0.705 0.889

Glutamic 0.971 1.022 0.776 0.923


amino acids

Proline 0.014 0.012 0.011 0.012

Glycine 0.901 0.880 0.708 0.83

Alanine 0.655 0.633 0.470 0.586

Tyrosine 0.394 0.406 0.276 0.359

Arginine 0.468 0.429 0.342 0.413

According to the second sample, that acquired the third date of


harvesting when laying workers percentage had 80%, the previously
mentioned non essential amino acid means were 2.439, 1.005, 1.022,
0.012, 0.880, 0.633, 0.406 and 0.429%, respectively,
In case of the third sample that harvested from the seventh
harvesting date where the laying workers percentage was 100%, the
prior non-essential amino acids obtained 1.900, 0.705, 0.776, 0.011,
0.708, 0.470, 0.276 and 0.342%, respectively.

161
For the time being laying workers mean percentage recorded
80%, the calculated general mean numbers of non-essential amino
acids were 2.285%, 0.889%, 0.923%, 0.012%, 0.830%, 0.586%,
0.359% and 0.413% for Aspartic, Serine, Glutamic, Proline, Glycine,
Alanine, Tyrosine and Arginine, respectively.
3. Carniolian hybrid used as production and donor colonies.
a. Essential amino acids
Results given in Table (38) showed that the first sample has
been taken in the first harvesting date when the percentage of laying
workers achieved 60%. In this case, the mean percentages of
Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine,
Histidine and Lysine acids were 0.529, 0.734, 0.107, 0.494, 0.965,
0.360, 0.343 and 0.986%, respectively.
Concerning the results of the second sample, which harvested in
the fourth harvesting date with 80% laying workers, the consecutive
means of essential amino acids were 0.481, 0.628, 0.051, 0.427, 0.809,
0.300, 0.270 and 0.895%, respectively.
Results of the third sample that attained in the sixth harvesting
date with 100% laying workers indicated that essential amino acids
recorded successively 0.496, 0.617, 0.044, 0.431, 0.834, 0.314, 0.291
and 0.804%, respectively.
The calculated general means of essential amino acids as laying
workers mean percentage had 80% were 0.502%, 0.660%, 0.067%,
0.451%, 0.869%, 0.325%, 0.301% and 0.895% for Threonine, Valine,
Methionine, Isoleucine, Leucine, Phenylalanine, Histidine and Lysine,
respectively.

162
Table 38. Mean percentages of essential amino acids in royal jelly
produced from different harvesting numbers and different
percentages of laying workers by Carniolian hybrid colonies
when used as production and donor colonies.

Sample No. 1 2 3
Mean
Harvesting No. 1st 4th 6th

Percentage of laying workers 60 80 100 80


Type and percentage of essential amino

Threonine 0.529 0.481 0.496 0.502

Valine 0.734 0.628 0.617 0.660

Methionine 0.107 0.051 0.044 0.067

Isoleucine 0.494 0.427 0.431 0.451


acids

Leucine 0.965 0.809 0.834 0.869

Phenylalanine 0.360 0.300 0.314 0.325

Histidine 0.343 0.270 0.291 0.301

Lysine 0.986 0.895 0.804 0.895

b. Non essential amino acids


Table (39) indicated that the first sample was taken in the 1st
harvesting date when the percentage of laying workers attained 60%. In
this sample, the percentages of Aspartic, Serine, Glutamic, Proline,
Glycine, Alanine, Tyrosine and Arginine acids were 2.412, 0.943,
1.025, 0.017, 0.842, 0.636, 0.389 and 0.432%, respectively.

163
Table 39. Mean percentages of essential amino acids in royal jelly
produced from different harvesting numbers and different
percentages of laying workers by Carniolian hybrid colonies
when used as production and donor colonies.
Sample No. 1 2 3
Mean
st th th
Harvesting No. 1 4 6

Percentage of laying workers 60 80 100 80

Aspartic 2.412 2.210 2.154 2.259


Type and percentage of non essential

Serine 0.943 0.914 0.896 0.918

Glutamic 1.025 0.972 0.935 0.977


amino acids

Proline 0.017 0.011 0.010 0.013

Glycine 0.842 0.824 0.790 0.819

Alanine 0.636 0.586 0.583 0.602

Tyrosine 0.389 0.339 0.341 0.356

Arginine 0.432 0.383 0.364 0.393

Regarding the second sample that obtained from the fourth


harvesting date with 80%laying workers, 2.210, 0.914, 0.972, 0.011,
0.824, 0.586, 0.339 and 0.383% were recorded for the respective
previously mentioned non-essential amino acids.
The third sample was obtained in the sixth harvesting date as
laying workers percentage was 100%. In this case, the percentages of
preceding amino acids reached 2.154, 0.896, 0.935, 0.010, 0.790,
0.583, 0.341 and 0.364%, respectively.

164
Furthermore, the same table denoted that the non-essential
amino acid averages were – in general – 2.259%, 0.918%, 0.977%,
0.013%, 0.819%, 0.602%, 0.356% and 0.393% for Aspartic, Serine,
Glutamic, Proline, Glycine, Alanine, Tyrosine and Arginine,
respectively when laying workers mean reached 80%.
In general view of results concerning amino acids, it can be
stated that there were no close relationship between amino acids values
(essential and non-essential) in different samples throughout different
harvesting dates and percentages of laying workers.
Examining results of essential amino acids illustrated in Table
(40), concerning the mean percentages and ranges of essential amino
acids in different samples which produced by Egyptian race, Italian and
Carniolian hybrids colonies from different harvesting numbers and
different percentages of laying workers when used as production and
donor colonies, the following points could be viewed;
In case of Threonine, the mean percentages varied in different
types of colonies where it reached 0.569, 0.509 and 0.502%, and
ranged between 0.500-0.625%, 0.424-0.562% and 0.481-0.529% for
Egyptian race, Italian hybrid and Carniolian hybrid, respectively.
Valine acid recorded 0.740, 672 and 0.660 %, and ranged
between 0.611-0.847%, 0.562-0.733% and 0.628-0.734% for
successive previous colonies.
Methionine had lower mean percentages (0.116, 0.111 and
0.067%) and ranged between 0.061-0.159%, 0.091-0.127% and 0.044-
0.107% for the above-mentioned colonies, respectively.

165
Table 40. General mean percentages of essential amino acids in royal
jelly produced by Egyptian race, Italian and Carniolian
hybrids colonies which represented different harvesting
numbers and different percentages of laying workers when
used as production and donor colonies.

Type of race and Carniolian


Egyptian raceItalian hybrid Average
hybrid hybrid
Percentage of laying 84 80 80
81
workers (60-100) (60-100) (60-100)
0.569 0.509 0.502
Type and percentage of essential amino

Threonine 0.527
(0.500-0.625) (0.424-0.562) (0.481-0.529)
0.740 0.672 0.660
Valine 0.691
(0.611-0.847) (0.562-0.733) (0.628-0.734)
0.116 0.111 0.067
Methionine 0.098
(0.061-0.159) (0.091-0.127) (0.044-0.107)
0.507 0.464 0.451
Isoleucine 0.474
(0.430-0.584) (0.389-0.514) (0.427-0.494)
acids

1.023 0.901 0.869


Leucine 0.931
(0.908-1.132) (0.746-1.000) (0.809-0.965)
0.402 0.338 0.325
Phenylalanine 0.355
(0.342-0.456) (0.267-0.378) (0.300-0.360)
0.382 0.313 0.301
Histidine 0.332
(0.320-0.463) (0.255-0.346) (0.270-0.343)
0.569 0.763 0.895
Lysine 0.742
(0.876-1.439) (0.644-0.844) (0.804-0.986)

Isoleucine had 0.507% for the Egyptian race colonies and


ranged from 0.430-0.584%, while it calculated 0.464% (0.389-0.514%)
in case of Italian hybrid colonies and in Carniolian hybrid bees
recorded 0.451% (0.427-0.494%).
Considering Leucine, it was averaged 1.023% for Egyptian race,
0.901% for Italian hybrid and 0.869% for Carniolian hybrid. These
percentages ranged 0.908-1.132%, 0.746-1.000% and 0.809-0.965%,
respectively.

166
Phenylalanine means varied between different types of colonies
were it reached 0.402, 0.338 and 0.325%, and ranged 0.342-0.456%,
0.267-0.378% and 0.300-0.360% for Egyptian race, Italian hybrid and
Carniolian hybrid colonies, respectively
Histadin had different percentages of 0.382, 0.313 and 0.301%
for Egyptian race, Italian hybrid and Carniolian hybrid colonies as it
ranged 0.320-0.4635%, 0.255-0.346% and 0.270-0.343%, respectively.
Mean percentage values of Egyptian race, Italian and Carniolian
hybrids colonies for Lysine were 0.569, 0.763 and 0.895% and ranged
between 0.876-1.439%, 0.644-0.844% and 0.804-0.986%, respectively.
When calculating the average mean percentage of essential
amino acids in royal jelly collected for different colonies as also shown
in Table (40), the resultant values were 0.527%, 0.691%, 0.098%,
0.474%, 0.931%, 0.355%, 0.332% and 0.742%, in support of
Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine,
Histidine and Lysine, respectively.
Obtained results for non-essential amino acids in royal jelly
produced in various harvesting dates from different colonies in which
different percentages of laying workers were used, are tabulated in
Table (41).
Aspartic acid recorded 2.302% for royal jelly produced from
Egyptian race colonies (ranged 1.331-2.733%), while it gave 2.285%
(1.00-2.516%) for Italian hybrid. It reached a percentage of 2.285%
which was in between of 1.00-2.516%. The same amino acid detected
in 2.259% of royal jelly produced from Carniolian hybrid colonies
where it varied from 2.154 and 2.412%.

167
Table 41. General mean percentages of non essential amino acids in
royal jelly produced by Egyptian race, Italian and Carniolian
hybrids colonies which represented different harvesting
numbers and different percentages of laying workers when
used as production and donor colonies.

Type of race and Carniolian


Egyptian race Italian hybrid Mean
hybrid hybrid
Percentage of laying 84 80 80
81
workers (60-100) (60-100) (60-100)
2.302 2.285 2.259
Aspartic 2.282
Type and percentage of non essential

(1.661-2.733) (1.900-2.516) (2.154-2.412)


1.007 0.889 0.918
Serine 0.938
(0.871-1.149) (0.705-1.005) (0.896-0.943)
1.068 0.923 0.977
Glutamic 0.989
(0.901-1.298) (0.776-1.022) (0.935-1.025)
amino acids

0.015 0.012 0.013


Proline 0.013
(0.008-0.022) (0.011-0.014) (0.010-0.017)
0.949 0.830 0.819
Glycine 0.866
(0.808-1.203) (0.708-0.901) 0.790-0.842
0.676 0.586 )(
0.602
Alanine 0.621
(0.588-0.784) (0.470-0.655) (0.583-0.636)
0.441 0.359 0.356
Tyrosine 0.385
(0.364-0.511) (0.276-0.406) (0.339-0.389)
0.480 0.413 0.393
Arginine 0.429
(0.408-0.562) (0.342-0.468) (0.364-0.432)

In the case of the produced Serine acid in royal jelly for


Egyptian race, Italian hybrid and Carniolian hybrid, it achieved 1.007,
0.889 and 0.918% with percentages ranged 0.871-1.149%, 0.705-
1.005% and 0.896-0.943%, respectively.
Results of Glutamic acid showed different percentages in
different colonies. Egyptian race colonies had 1.068% that ranged

168
between 0.901 and 1.298%. Italian and Carniolian hybrids gave
0.923% (0.776-1.022%) and 0.977% (0.935-1.025%), respectively.
Proline acid reached 0.015, 0.012 and 0.013% in the Egyptian
race, Italian hybrid and Carniolian hybrid colonies in which the same
amino acid ranged 0.008-0.022%, 0.011-0.014% and 0.010-0.017%,
consecutively.
In addition, Glycine percentages varied according to the type of
colonies. It achieved 0.949% in Egyptian race colonies while it reached
0.830% in Italian hybrid ones. As for Carniolian hybrid, the royal jelly
had a percentage of 0.819. In these colonies, the produced royal jelly
ranged 0.808-1.203%, 0.708-0.901% and 0.790-0.842%, respectively.
Alanine percentages were 0.676, 0.586 and 0.602% where they
ranged 0.588-0.784%, 0.470-0.655% and 0.583-0.636% for Egyptian
race, Italian and Carniolian hybrids colonies, respectively.
Percentages of Tyrosine reached 0.441, 0.359 and 0.356% for
Egyptian race, Italian and Carniolian hybrids colonies, respectively.
These percentages varied with the type of colony as it varied
successively from 0.364 to 0.511%, 0.276 to 0.406% and 0.339 to
0389%.
In case of Arginine, 0.480% was recorded for Egyptian race,
0.413% for Italian hybrid and 0.393% for Carniolian hybrid. These
percentages ranged 0.408-0.562%, 0.342-0.468% and 0.364-0.432%,
respectively.
The general mean percentages of non essential amino acid rates
in royal jelly in different colonies when 0.385% were as follows;
2.282% for Aspartic, 0.938% for Serine, 0.989% for Glutamic, 0.013%

169
for Proline, 0.866% for Glycine, 0.621% for Alanine, 0.385% for
Tyrosine and 0.385% for Tyrosine.
It could generally concluded from results of essential amino
acids that the maximum values were attained in Egyptian race samples
followed consecutively in Italian hybrid and Carniolian hybrid samples
with exception of Lysine acid. This essential amino acid gave the
highest value in Carniolian hybrid then Italian hybrid and finally
Egyptian race.
Results of non-essential amino acids revealed the occurrence
of maximum values in the Egyptian race samples. The second and third
ranks oscillated between Italian hybrid and Carniolian hybrid samples.
The resulting amino acid percentages that produced from royal
jelly samples were generally low as compared with those collected in
the same apiary yard in Giza region since 35 years ago. In this respect,
Eweis (1974) stated that the fresh royal jelly had higher nitrogen
content whereas the percentages of amino acids were 3.95, 1.41, 3.62,
1.59, 2.09, 3.32, 5.22 and 1.81% for Threonine, Valine, Methionine,
Isoleucine, Leucine, Phenylalanine, Histidine and Lysine, respectively.
Those differences in the percentages of amino acids could be
referred to many reasons such as the sharp reduction of area cultivated
with plants, bees keeping that mainly depended on artificial feeding. In
addition, the ecological factors which play an important role on the
production colonies. Furthermore, the genetic origin of production
colonies may be clearly affect the quality of royal jelly.

170
b. Determination of fatty acid percentages
Royal jelly analysis for fatty acids showed variations between
the mean percentages for the same fatty acid in different samples
during different harvesting dates and laying workers percentages.
1. Egyptian race used as production and donor colonies
Analysis of these colonies revealed the incidence of five fatty
acids in all tested samples with exception of the first and second ones
in which five and seven fatty acids were detected (Table 42).
In the first sample (collected in the first harvesting date), where
60% of laying workers happened, the detected five fatty acids were as
follows; 10-hydroxy-2-decenoic acid (10HDA) which found in
relatively higher percentage (3.158%). This fatty acid was followed in
descending orders with Cis,cis-9,12-Octadecadienoic (0.333%),
Tetracosanoic (0.248%), Eicosanoic (0.167%) and Hexadecanoic
(0.094%).
As for the second sample that obtained from the second
harvesting date when the laying workers percentage was 80%, the
results showed the occurrence of seven fatty acids. The highest
percentage noticed in 10-hydroxy-2-decenoic acid (10HDA) where
2.731% detected. The other six fatty acids were Hexadecanoic,
Octadecanoic, Cis,cis-9,12-Octadecadienoic, Eicosanoic,
Tetracosanoic, and Cis-9-Octadecenoic acids with 0.097, 0.122, 0.040,
0.394, 0.053 and 0.647%, respectively.
The third sample was taken from the 3rd harvesting date when
85% of laying workers took place. The mean percentages of
Hexadecanoic, Octadecanoic, Cis,cis-9,12-Octadecadienoic,

171
Eicosanoic, Tetracosanoic, and Cis-9-Octadecenoic acids were 0.170,
0.000, 0.729, 0.361, 0.169 and 0.063% respectively. The mean
percentage of 10-hydroxy-2-decenoic acid (10HDA) was the highest
(2.392%).

Table 42. Mean percentages of fatty acids in royal jelly produced from
different harvesting numbers and percentages of laying
workers by Egyptian race colonies when used as production
and donor colonies.

Sample No. 1 2 3 4 5
Mean
Harvesting No. 1st 2nd 3rd 4th 5th
Percentage of laying
60 80 85 95 100 84
workers
10-hydroxy-2-
3.158 2.482 2.392 2.857 2.765 2.731
Type and percentage of fatty acids

decenoic acid
(10HDA)
Hexadecanoic acid 0.094 0.097 0.170 0.114 0.091 0.113

Octadecanoic acid 0.000 0.122 0.000 0.000 0.000 0.024


Cis,cis-9,12-
0.333 0.040 0.729 0.546 0.481 0.426
Octadecadienoic
acid
Eicosanoic acid 0.167 0.394 0.361 0.324 0.260 0.301
Tetracosanoic
0.248 0.053 0.169 0.028 0.298 0.159
acid
Cis-9-
0.000 0.647 0.063 0.052 0.047 0.162
Octadecenoic acid

Concerning the fourth sample which gained from the fourth date
with 95% laying workers, the mean percentages of Hexadecanoic,
Octadecanoic, Cis,cis-9,12-Octadecadienoic, Eicosanoic,
Tetracosanoic, and Cis-9-Octadecenoic acids were 0.114, 0.000, 0.546,

172
0.324, 0.028 and 0.052%, respectively. The 10-hydroxy-2-decenoic
acid (10HDA) demonstrated highest percentage (2.857%).
In the case of the fifth sample, where 100% of laying workers
took place, Hexadecanoic, Cis,cis-9,12-Octadecadienoic, Eicosanoic,
Tetracosanoic, and Cis-9-Octadecenoic acids gave 0.091, 0.481, 0.260,
0.298 and 0.047%, respectively. The percentage of 10-hydroxy-2-
decenoic acid (10HDA) reached 2.765%.
The general means of fatty acids - in which the mean percentage
of laying workers was 84% - represented the following; 0.113%,
0.024%, 0.426%, 0.301%, 0.159%, and 0.162% for Hexadecanoic,
Cis,cis-9,12-Octadecadienoic, Eicosanoic, Tetracosanoic, and Cis-9-
Octadecenoic acids. The mean of 10-hydroxy-2-decenoic acid
(10HDA) was the maximum with 2.731%.
2. Italian hybrid used as production and donor colonies.
Table (43) revealed that number of fatty acids varied in the
different tested samples. The first sample had four acids, while the
second one recorded six acids. At the same time, the third sample had
seven acids.
In the of first sample which obtained from the first harvesting
date as laying workers was 60%, the mean percentages of
Hexadecanoic, Cis,cis-9,12-Octadecadienoic and Eicosanoic were
0.068, 0.290 and 0.117%, respectively. The mean percentage of 10-
hydroxy-2-decenoic acid (10HDA) was 3.347%.
According to presented data of the second sample which
harvested from the third date of harvesting, the detected six fatty acids
were 10-hydroxy-2-decenoic acid (10HDA), Hexadecanoic,

173
Octadecanoic, Eicosanoic, Tetracosanoic and Cis-9-Octadecenoic. The
percentages of preceding acids were 2.817, 0.128, 0.049, 0.347, 0.033
and 0.572%, respectively.
Table 43. Mean percentages of fatty acids in royal jelly produced from
different harvesting numbers and percentages of laying
workers by Italian hybrid colonies when used as production
and donor colonies.

Sample No. 1 2 3
Mean
st th th
Harvesting No. 1 4 7

Percentage of laying workers 60 80 100 80


10-hydroxy-2-
3.347 2.817 2.156 2.773
Type and percentage of fatty acids

decenoic acid
(10HDA)
Hexadecanoic acid 0.068 0.128 0.143 0.113

Octadecanoic acid 0.000 0.049 0.099 0.049


Cis,cis-9,12-
0.290 0.000 0.068 0.119
Octadecadienoic
acid
Eicosanoic acid 0.117 0.347 0.298 0.254

Tetracosanoic acid 0.000 0.033 0.444 0.159


Cis-9-Octadecenoic
0.000 0.572 0.546 0.373
acid

In the third sample which harvested from the seventh date with
100% of laying workers, the seven fatty acids were 10-hydroxy-2-
decenoic acid (10HDA), Hexadecanoic, Octadecanoic, Cis,cis-9,12-
Octadecadienoic, Eicosanoic, Tetracosanoic, and Cis-9-Octadecenoic.
The consecutive mean percentages of afore acids recorded 2.156,
0.143, 0.099, 0.068, 0.298, 0.444 and 0.546%.

174
The general means of fatty acids were calculated from the
previous percentages which representing the following;
10-hydroxy-2-decenoic acid (10HDA) with 2.773%,
Hexadecanoic with 0.113%, Octadecanoic with 0.049%, Cis,cis-9,12-
Octadecadienoic with 0.119%, Eicosanoic with 0.254%, Tetracosanoic
with 0.159%, and Cis-9-Octadecenoic with 0.373%.
3. Carniolian hybrid used as production and donor colonies.
Table (44) illustrated that the detected fatty acids varied in their
numbers in the tested three samples where the first sample had five
acids, while both the second and third ones recorded seven acids.
The first sample taken from the first harvesting date, as the
percentage of laying workers was 60%. The mean percentages of
Hexadecanoic, Eicosanoic, Tetracosanoic, and Cis-9-Octadecenoic
acids were 0.079, 0.087, 0.310 and 0.204%, respectively. The 10-
hydroxy-2-decenoic acid (10HDA) gave the highest percentage with
2.392%.
According to presented data, the second sample was harvested
from the fourth date of harvesting with laying workers percentage of
80%. The seven fatty acids were 10-hydroxy-2-decenoic acid
(10HDA), Hexadecanoic, Octadecanoic, Cis,cis-9,12-Octadecadienoic,
Eicosanoic, Tetracosanoic and Cis-9-Octadecenoic. The respective
means of earlier fatty acids were 2.575, 0.216, 0.026, 0.567, 0.336,
0.090 and 0.116%, respectively.
In case of the third sample that attained from the sixth
harvesting date when laying workers percentage recorded 100%, the
previous seven fatty acids detected in the second sample were occurred.

175
The successive mean percentages recorded 1.132, 0.225, 0.241,
0.101, 0.588, 0.086 and 0.894%.

Table 44. Mean percentages of fatty acids in royal jelly produced from
different harvesting numbers and percentages of laying
workers by Carniolian hybrid colonies when used as
production and donor colonies.

Sample No. 1 2 3
Mean
st rd th
Harvesting No. 1 3 6

Percentage of laying workers 60 80 100 80


10-hydroxy-2-decenoic
2.392 2.575 1.132 2.033
Type and percentage of fatty acids

acid (10HDA)
Hexadecanoic acid 0.079 0.216 0.225 0.173

Octadecanoic acid 0.000 0.026 0.241 0.089


Cis,cis-9,12-
0.000 0.567 0.101 0.223
Octadecadienoic acid
Eicosanoic acid 0.087 0.336 0.588 0.337

Tetracosanoic acid 0.310 0.090 0.086 0.162


Cis-9-Octadecenoic
0.204 0.116 0.894 0.405
acid

Considering the general means of fatty acids, they calculated


2.033%, 0.173%, 0.089%, 0.223%, 0.337%, 0.162%, and 0.405% for
10-hydroxy-2-decenoic acid (10HDA), Hexadecanoic, Octadecanoic,
Cis,cis-9,12-Octadecadienoic, Eicosanoic, Tetracosanoic and Cis-9-
Octadecenoic, respectively.
According to the results obtained, it is evident that there were no
differences among the tested samples represented different harvesting

176
dates. In addition, the percentages of laying workers had no effect on
fatty acid values.
The calculated general percentage means of fatty acids identified
in royal jelly yielded from different harvesting dates for Egyptian race,
Italian and Carniolian hybrids colonies that represented different
percentages of laying workers when used as production and donor
colonies shown in Table (45).
Consideration of results obtained for royal jelly produced from
Egyptian race colonies, it may be of interest to mention that 10-
hydroxy-2-decenoic acid (10HDA) gave the highest percentage where
it averaged 2.731% and was in between of 2.392-3.158%.
This fatty acid was followed successively with Cis,cis-9,12-
Octadecadienoic, Eicosanoic, Cis-9- Octadecenoic, Tetracosanoic,
Hexadecanoic and Octadecanoic. The calculated mean percentages and
ranges of these acids were 0.426 (0.040-0.729%), 0.301 (0.167-
0.394%), 0.162 (0.000-0.647%), 0.159 (0.028-0.298%), 0.113 (0.091-
0.170%) and 0.024 (0.000-0.122%), respectively.
Fatty acids distinguished in royal jelly obtained from Italian
hybrid colonies reached mean percentages of 0.113, 0.049, 0.119,
0.254, 0.159 and 0.373% for Hexadecanoic, Octadecanoic, Cis,cis-
9,12- Octadecadienoic, Eicosanoic, Tetracosanoic and Cis-9-
Octadecenoic acids, respectively. The corresponding percentages
ranged between 0.068-0.143%, 0.000-0.099%, 0.000-0.290%, 0.117-
0.347%, 0.000-0.444% and 0.000-0.572%. As for 10-hydroxy-2-
decenoic acid (10HDA), it gave 2.773% and ranged 2.156-3.347%.

177
Carniolian hybrid's royal jelly had mean percentages of 0.173,
0.089, 0.223, 0.337, 0.162 and 0.405% for Hexadecanoic,
Octadecanoic, Cis,cis-9,12- Octadecadienoic, Eicosanoic,
Tetracosanoic and Cis-9-Octadecenoic, respectively. Previous means
had an intermediate percentages ranged 0.079-0.225, 0.000-0.241,
0.000-0.567, 0.087-0.588, 0.087-0.310 and 0.116-0.894%, respictively.
The 10-hydroxy-2-decenoic acid (10HDA) percentage was 2.033% that
ranged from 2.575 to 1.132%.

Table 45. General mean percentages of fatty acids in royal jelly


produced by Egyptian race, Italian and Carniolian hybrids
colonies which represented different harvesting numbers and
different percentages of laying workers when used as
production and donor colonies.
Genetic origin of Carniolian
Egyptian race Italian hybrid Mean
colony hybrid
Percentage of 84 80 80
81
laying workers (60-100) (60-100 ) (60-100)
10-hydroxy-2-
2.731 2.773 2.033
decenoic acid 2.512
(2.392 - 3.158) (2.156 - 3.347) (1.132-2.575)
Type and percentage of fatty acids

(10HDA)
Hexadecanoic 0.113 0.113 0.173
0.133
acid (0.091 - 0.170) (0.068 - 0.143) (0.079 - 0.225)
Octadecanoic 0.024 0.049 0.089
0.054
acid (0.000 - 0.122) (0.000 - 0.099) (0.000 - 0.241)
Cis,cis-9,12-
0.426 0.119 0.223
Octadecadienoic 0.256
(0.040 - 0.729) (0.000 – 0.290) (0.000 - 0.567)
acid
0.301 0.254 0.337
Eicosanoic acid 0.297
(0.167 - 0.394) (0.117 - 0.347) (0.087 - 0.588)
Tetracosanoic 0.159 0.159 0.162
0.160
acid (0.028 - 0.298) (0.000 – 0.444) (0.086 -0.310)
Cis-9-
0.162 0.373 0.405
Octadecenoic 0.313
(0.000 - 0.647) (0.000 – 0.572) (0.116 -0.894)
acid

178
The seven fatty acids could be arranged in descending orders
according to their general percentage means as follows; 10-hydroxy-2-
decenoic (10HDA), Cis-9-Octadecenoic, Eicosanoic, Cis,cis-9,12-
Octadecadienoic, Tetracosanoic, Hexadecanoic and Octadecanoic. The
general averages of the previous fatty acids were 2.512% (1.132-
2.575), 0.313% (0.116 -0.894), 0.297% (0.087 - 0.588), 0.256% (0.000
- 0.567), 0.160% (0.086 -0.310), 0.133% (0.079 - 0.225) and .054%
(0.000 - 0.241), respectively.
These results are in accordance with the results of Genc and
Aslan (1999). They analyzed several royal jellies and commercial
products containing royal jelly for their 10-hydroxy-2-decenoic acid
(10-HDA) content using a column liquid chromatography technique
and found that 10-HDA mean percentage was in between 0.75 and
2.54%.
In the same respect, Zhou et al. (2007) used an ultra
performance liquid chromatography (UPLC) and high performance
liquid chromatography (HPLC) method to detect the 10-hydroxy- 2-
decenoic acid (10-HDA) content in royal jelly cream and lyophilized
powder. The concentration of 10-HDA ranges from 1.26 to 2.25% for
pure royal jelly cream samples and 3.01–6.26% for royal jelly
lyophilized powder samples. For 30 royal jelly products, the 10-HDA
content varied from no detectable to 1.005%.
According to obtained results, it could be conclud that although
fatty acids gave different percentages, they reached their maximum
values in most cases in the Carniolian hybrid samples, whereas the
second and third ranks fluctuated between Italian hybrid and Egyptian

179
race samples with the exception of both 10-hydroxy-2-decenoic acid
(10HDA) and Cis,cis-9,12- Octadecadienoic. As for the first fatty acid,
the Italian hybrid samples had the highest amount followed by
Egyptian race then the Carniolian hybrid. In case of the second fatty
acid, the maximum value was detected in the Egyptian race samples
then Carniolian and Italian hybrids, respectively.

180
SUMMARY
Royal jelly is one of the most valued products of honeybee
colonies, and produced from hypopharyngeal and mandibular glands of
6-12 days old workers, which called nurse bees.
This work was carried out in the apiary yard of the Agricultural
Experimental Station, Faculty of Agriculture, Cairo University, Giza
Governorate in late spring and summer of 2008 and 2009.
Obtained results can be summarized as follows:
1. Evaluation of some factors affecting royal jelly production in
honeybee colonies
Experiments of this study are separated into colonies grafted
with non-related larvae and colonies grafted with related grafted larvae.
Royal jelly could be harvested from each studied hybrid or race each 3
days and continued until appearance of laying workers, thus number of
harvesting dates varied.
a. None related grafted larvae
This research was conducted using the Egyptian race and
Carniolian hybrid queenless production colonies that grafted with
different genetic origin larvae.
Concerning Egyptian race colonies grafted with different genetic
origin larvae throughout the period extended from 22/3/2008 to
31/3/2008, the results showed that the first bar gave the highest
percentage of acceptance and amount of RJ, whereas the second one
had an intermediate acceptance percentage and lowest amount of RJ,
but the third bar showed the lowest percentage of acceptance with an
intermediate RJ amount. The mean percentage of acceptance was

181
51.9%, whereas the mean amount of harvested royal jelly was 0.438
gm. The effect of accepted grafted larvae percentage on the amount of
harvested royal jelly as well as the influence of each of laying workers
percentage and number of combs covered with bees (C.S.) on both
acceptance percentage and amount of harvested royal jelly were
insignificant.
In the Carniolian hybrid colonies grafted with Carniolian race
throughout the period extended from 21/4/2008 to 30/4/2008, the
results clarified that the highest percentage of acceptance and amount
of RJ occurred in the first bar followed by the second one, whereas the
third bar recorded the lowest values. These colonies averaged 54.7%
and 1.103 gm for percentage of acceptance and harvested royal jelly,
respectively. An insignificant effect was recorded for accepted grafted
larvae percentage on amount of harvested royal jelly. In each of the
relation between laying workers percentage and number of combs
covered with bees (C.S.), the influence was highly significant for the
accepted grafted larvae while it was insignificant for the harvested
royal jelly.
Results of Carniolian hybrid colonies grafted with different
genetic origin larvae throughout the period extended from 4/7/2008 to
19/7/2008 denoted that the minimum percentage of acceptance and
amount of RJ took place in the first bar, but the highest values occurred
in the second one, while intermediary values were recorded for the
third bar. In these colonies, the means of acceptance percentage and
amount of harvested royal jelly were 48.1% and 1.141 gm,
respectively. The relation between accepted grafted larvae percentage

182
and amount of harvested royal jelly was insignificant. In addition,
insignificant values were recorded for the effect of laying workers
percentage and number of combs covered with bees (C.S.) on both
acceptance percentage and amount of harvested royal jelly.
With reference to results of Carniolian hybrid colonies grafted
with different genetic origin larvae throughout the period extended
from 6/8/2008 to 15/8/2008, the minimum percentage of acceptance
and amount of RJ were proved for the first bar, while the second one
had the highest values though the third bar gave medial values. These
colonies calculated 57.7% for percentage of acceptance and 0.820 gm
for amount of harvested royal jelly. Relationship between accepted
grafted larvae percentage and amount of harvested royal jelly was
insignificant. In addition, the influences of laying workers percentage
on both acceptance percentage and amount of harvested royal jelly
were insignificant. As for the effects of number of combs covered with
bees (C.S.) on both acceptance percentage and amount of harvested
royal jelly, they were significant.
Regarding the Egyptian race colonies grafted with different
genetic origin larvae throughout the period extended from 22/3/2009 to
6/4/2009, obtained results revealed that the first bar gave the maximum
percentage of acceptance and amount of RJ, followed by the second
one whereas the third bar recorded the lowest values. The means of
these colonies were 35.4% for acceptance percentage and 0.519 gm for
amount of harvested royal jelly. The effect of accepted grafted larvae
percentage on amount of harvested royal jelly was highly significant. In
addition, the effect of each laying workers percentage and number of

183
combs covered with bees (C.S.) on both acceptance percentage and
amount of harvested RJ were also highly significant.
As general conclusions of non-related grafted larvae, the highest
mean percentage of acceptance occurred in the second bar (55.2%),
while the lowest rate detected in the third one (42.8%). The first bar
had an intermediate mean percentage of 50.4%. In case of amount of
harvested royal jelly, the greatest mean was amount collected from the
first bar, followed by the second one, then the third bar (0.841, 0.836
and 0.701%, respectively).
The average of acceptance percentage for these colonies was
49.6% while the average amount of harvested royal jelly was 0.804 gm.
The majority of previous colonies indicated insignificant
relationships between the accepted grafted larvae percentage and
amount of harvested royal jelly. Each laying workers percentage and
number of combs covered with bees (C.S.) indicated insignificant
values on both acceptance percentage and amount of harvested royal
jelly, with the exception of the fifth one (Egyptian race) which was
occurred in the period of 22/3/2009 to 6/4/2009 as it showed highly
significant relations.
b. Related grafted larvae
This study was conducted using the Egyptian race, Italian and
Carniolian hybrids queenless production colonies, which used as
production and donor colonies as follows:
In respect of Egyptian race colonies used as production and
donor colonies throughout the period extended from 8/4/2009 to
20/4/2009, the first bar gave the minimum percentage of acceptance

184
and amount of RJ but the third one had the highest percentage and
amount, whereas the second bar recorded intermediate values. The
mean percentage of acceptance recorded 32.5%, whilst the mean
amount of harvested royal jelly was 0.978 gm. In case of the effect of
accepted grafted larvae percentages on amount of harvested royal jelly,
the resultant value was significant. Furthermore, insignificant effects
were recorded for each of laying workers percentage and number of
combs covered with bees (C.S.) on both acceptance percentage and
amount of harvested royal jelly.
In relation to Italian hybrid colonies used as production and
donor colonies throughout the period extended from 15/7/2009 to
2/8/2009, the highest percentage of acceptance and amount of RJ were
occurred in the third bar, but the first bar had the lowest values,
whereas the second bar recorded intermediate acceptance and the
amount of RJ. These colonies gave 64.6% for acceptance percentage
mean and 0.959 gm for harvested royal jelly mean. The accepted
grafted larvae percentage had an insignificant influence on the amount
of harvested royal jelly. In the same trend, the effects of each laying
workers percentage and number of combs covered with bees (C.S.) on
both acceptance percentage and amount of harvested royal jelly were
also insignificant.
As for Carniolian hybrid colonies used as production and donor
colonies throughout the period extended from 13/8/2009 to 28/8/2009,
the results clarified that the first bar gave the minimum percentage of
acceptance but the third one had the highest rate, while the second bar
recorded an intermediate value. On the other hand, the second bar had

185
the maximum amount of harvested RJ, followed by the first one then
the third bar. The mean percentage of acceptance calculated 70.2%,
while the mean amount of harvested royal jelly was 1.535 gm. Highly
significant value was obtained for the effect of accepted grafted larvae
percentage on amount of harvested royal jelly. Reversely, insignificant
values were attained for the effects of each laying workers percentage
and number of combs covered with bees (C.S.) on both acceptance
percentage and amount of harvested royal jelly.
As a conclusion of related grafted larvae, the highest percentage
of acceptance and amount of harvested royal jelly means were occurred
in the third bar (65.6% and 1.333 gm, respectively), but the lowest
values were detected in the first one (44.8% and 0.905 gm,
respectively), whereas the second bar had an intermediate values
(56.8% and 0.905 gm, respectively.
The percentage of acceptance for these colonies averaged
55.8%, while the amount of harvested royal jelly averaged 1.157%.
Insignificant relationships were recorded for all tested colonies
between accepted grafted larvae the percentage on amount of harvested
royal jelly. Furthermore, the influences of each laying workers
percentage and number of combs covered with bees (C.S.) separately
on both acceptance percentage and amount of harvested royal jelly
were also insignificant
As a final conclusion from previous results concerning the
evaluation of some factors affecting royal jelly production, colonies
grafted with related larvae had highest means of acceptance percentage
(55.8%) and amount of harvested royal jelly (1.157 gm) than colonies

186
grafted with non-related larvae in which 49.6% and 0.804 gm were
recorded, respectively.
2. Determination of amino and fatty acid percentages in some
samples of the produced royal jelly
a. Determination of amino acid percentages
Results pertaining Egyptian race used as production and donor
colonies when laying workers mean percentage was 71% indicated that
the general means of essential amino acids were 0.569, 0.740, 0.116,
0.507, 1.023, 0.402, 0.382 and 0.569% for Threonine, Valine,
Methionine, Isoleucine, Leucine, Phenylalanine, Histidine and Lysine,
respictively. In case of non-essential amino acids, the general means
recorded 2.230, 1.007, 1.068, 0.015, 0.949, 0.676, 0.441 and 0.480%
for Aspartic, Serine, Glutamic, Proline, Glycine, Alanine, Tyrosine and
Arginine, respectively.

When laying workers percentage averaged 80% in the Italian


hybrid used as production and donor colonies, the means of essential
amino acids recorded 0.509, 0.672, 0.111, 0.464, 0.901, 0.338, 0.313
and 0.763% for Threonine, Valine, Methionine, Isoleucine, Leucine,
Phenylalanine, Histidine and Lysine, respictively. The calculated
general mean numbers of non-essential amino acids were 2.285, 0.889,
0.923, 0.012, 0.830, 0.586, 0.359 and 0.413% for Aspartic, Serine,
Glutamic, Proline, Glycine, Alanine, Tyrosine and Arginine,
respectively.

Regarding Carniolian hybrid used as production and donor


colonies when laying workers mean percentage calculated 80%, the

187
general means of essential amino acids were 0.502, 0.660, 0.067, 0.451,
0.869, 0.325, 0.301 and 0.895% for Threonine, Valine, Methionine,
Isoleucine, Leucine, Phenylalanine, Histidine and Lysine, respectively.
The non-essential amino acids averaged 2.259, 0.918, 0.977, 0.013,
0.819, 0.602, 0.356 and 0.393% for Aspartic, Serine, Glutamic, Proline,
Glycine, Alanine, Tyrosine and Arginine, respectively.
In a conclusion for amino acid percentages, no close
relationships were appeared between amino acid values (essential and
non-essential) in different samples throughout different harvesting
dates and different percentages of laying workers. The maximum
values of essential amino acids took place in Egyptian race samples
followed consecutively by Italian hybrid and Carniolian hybrid
samples, with an exception of Lysine acid. This essential amino acid
gave the highest value in Carniolian hybrid, followed by Italian hybrid
then Egyptian race. The maximum values for non-essential amino
acids occurred in the Egyptian race samples, whereas the second and
third ranks oscillated between Italian hybrid and Carniolian hybrid
samples.
b. Determination of fatty acid percentages
The general means of fatty acids in Egyptian race used as
production and donor colonies when laying workers mean was 84%
represented the following values; Hexadecanoic with 0.113%,
Octadecanoic had 0.024%, Cis,cis-9,12-Octadecadienoic gave 0.426%,
Eicosanoic proved 0.301%, Tetracosanoic had 0.159%, and Cis-9-
Octadecenoic with 0.162%. The mean of 10-hydroxy-2-decenoic acid
(10HDA) was the maximum with 2.731%.

188
According to the results of Italian hybrid used as production and
donor colonies, the calculated fatty acids general means represented the
following values 2.773, 0.113, 0.049, 0.119, 0.254, 0.159, and 0.373%
for 10-hydroxy-2-decenoic acid (10HDA), Hexadecanoic,
Octadecanoic, Cis,cis-9,12-Octadecadienoic, Eicosanoic, Tetracosanoic
and Cis-9-Octadecenoic acids, respectively.
Results of Carniolian hybrid used as production and donor
colonies showed that fatty acids calculated 2.033, 0.173, 0.089, 0.223,
0.337, 0.162, 0.405% for 10-hydroxy-2-decenoic acid (10HDA),
Hexadecanoic, Octadecanoic, Cis,cis-9,12-Octadecadienoic,
Eicosanoic, Tetracosanoic and Cis-9-Octadecenoic, respectively.
Results of fatty acids lead to conclude that they reached their
maximum values in the Carniolian hybrid samples, whereas the second
and third ranks fluctuated between Italian hybrid and Egyptian race
samples with an exception of both 10-hydroxy-2-decenoic acid
(10HDA) and Cis,cis-9,12- Octadecadienoic. As for the first fatty acid,
the Italian hybrid samples had the highest amount followed by
Egyptian race then the Carniolian hybrid. In case of the second fatty
acid, the maximum value was detected in the Egyptian race samples
then the Carniolian and Italian hybrids, respectively.

189
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‫الملخص العزتي‬

‫دراسات على إنتاج الغذاء الملكي في طىائف نحل العسل‬

‫‪ُٝ‬عرثش اىغزاء اىَين‪ ٍِ ٜ‬إٌٔ ٍْرعاخ غ٘ائف ّحو اىعغو ‪ٗ 0‬ذقً٘ شغاالخ ّحو اىعغو‬
‫اىر‪ ٜ‬ذثيغ ٍِ اىعَش ‪ ً٘ٝ 12 -6‬تإّراض اىغزاء اىَين‪ ،ٜ‬ح‪ٞ‬س ‪ُٝ‬فشص ٍِ اىغذد ذحد اىثيعٍ٘‪ٞ‬ح‬
‫ٗاىفن‪ٞ‬ح‪ٗ ،‬ذُغَ‪ٕ ٚ‬زٓ اىشغاالخ "تاىشغاالخ اىحاظْح" ‪0‬‬
‫اىضساع‪ٞ‬ح – مي‪ٞ‬ح اىضساعح – ظاٍعح‬ ‫ذٌ إظشاء ٕزٓ اىثحس تَْحو ٍحطح اىرعاسب‬
‫اىقإشج ف‪ٍ ٜ‬حافظح اىع‪ٞ‬ضج خاله فصي‪ ٜ‬اىشت‪ٞ‬ع ٗاىص‪ٞ‬ف ٍِ عاٍ‪02009 -2008 ٜ‬‬
‫ويُمكن تلخيص النتائج المتحصل عيها كما يلي‪:‬‬
‫‪ .1‬تقييم تعط العىامل المؤثزة على إنتاج الغذاء الملكي تطىائف نحل العسل‬
‫ذٌ ذقغ‪ ٌٞ‬اىرعاسب اىخاصح تٖزٓ اىذساعح إى‪ ٚ‬غ٘ائف ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ أصو‬
‫‪ٗ 0‬قذ ذٌ‬ ‫ٗساش‪ٍ ٜ‬خريف عْٖا ٗ غ٘ائف ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ ٜ‬ىٖا‬
‫حصاد اىغزاء اىَين‪ ٍِ ٜ‬مو ٕع‪ ِٞ‬أٗ عالىح مو ‪ 3‬أ‪ٝ‬اً حر‪ ٚ‬ظٖ٘س األٍٖاخ اىنارتح‪ٍَ ،‬ا أد‪ٙ‬‬
‫إى‪ ٚ‬اخرالف عذد ٍشاخ اىحصاد ‪0‬‬
‫أ‪ .‬طىائف تم تطعيمها تيزقات من أصل وراثي مختلف عنها‬
‫ذٌ إظشاء ٕزٓ اىذساعح ت٘اعطح غ٘ائف اإلّراض اىخاصح تغالىح اىْحو اىَصش‪ ٛ‬اىْق‪ٜ‬‬
‫ٗاىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬عذ‪َٝ‬ح اىَيناخ اىالئ‪ ٜ‬ذٌ ذطعَ‪ ٔٞ‬ا ت‪ٞ‬شقاخ ٍخريفح األصو اى٘ساش‪ ٜ‬عِ‬
‫األصو اى٘ساش‪ ٜ‬ىطائفح اإلّراض ‪0‬‬
‫ف‪ ٜ‬حاىح غ٘ ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍخريفح األصو‬
‫اى٘ساش‪ ٜ‬خاله اىفرشج ٍِ ‪ 2008 /3/22‬إى‪ ،2008 /3/31 ٚ‬أظٖشخ اىْرائط أُ اىغُذاتح األٗى‪ٜ‬‬
‫ف‪ ٜ‬ح‪ ِٞ‬أّرعد اىغُذاتح اىصاّ‪ٞ‬ح ّغثح‬ ‫أعطد أعي‪ّ ٚ‬غثح قث٘ه ٗأمثش مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ٜ‬‬
‫ٍر٘عطح ٍِ اىقث٘ه ٗأقو مَ‪ٞ‬ح غزاء ٍين‪ ، ٜ‬ف‪ّ ٛ‬فظ اى٘قد فقذ ماُ ىيغُذاتح اىصاىصح أقو ّغثح‬
‫‪% 51,9‬‬ ‫قث٘ه ٗمَ‪ٞ‬ح ٍر٘عطح ٍِ اىغزاء اىَين‪ 0 ٜ‬ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف‬
‫‪ 0,438‬ظٌ ‪ 0‬ماُ ذأش‪ٞ‬ش ّغثح قث٘ه اى‪ٞ‬شقاخ‬ ‫ت‪َْٞ‬ا ماُ ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‬
‫مزىل ماُ ىرأش‪ٞ‬ش مو ٍِ األٍٖاخ‬ ‫اىَطعٍ٘ح عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪،ٛ‬‬

‫‪1‬‬
‫اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‬
‫أ‪ٝ‬عا ً غ‪ٞ‬ش ٍعْ٘‪0 ٛ‬‬
‫تاىْغثح هغ٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ أصو مشّ‪٘ٞ‬ى‪ّ ٜ‬ق‪ٜ‬‬
‫خاله اىفرشج ٍِ ‪ 2008 /4/21‬إى‪ ، 2008 /4/30 ٚ‬فيقذ أٗظحد اىْرائط أُ أعي‪ّ ٚ‬غثح ٍِ‬
‫اهقث٘ه ٗأمثش مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ ٜ‬حذشد ف‪ ٜ‬اىغُذاتح األٗى‪ ٚ‬ذيرٖا اىغُذاتح اىصاّ‪ٞ‬ح ف‪ ٜ‬ح‪ِٞ‬‬
‫ععيد اىغُذاتح اىصاىصح أقو ّغثح قث٘ه ٗمَ‪ٞ‬ح غزاء ٍين‪ 0 ٜ‬ماُ ٍر٘عػ ٕزٓ اىط٘ائف ‪ٗ ٪ 54,7‬‬
‫‪ 1,103‬ظٌ ىنو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ، ٜ‬عي‪ ٚ‬اىر٘اى‪ 0 ٜ‬ماُ ذأش‪ٞ‬ش ّغثح قث٘ه‬
‫اى‪ٞ‬شقاخ اىَطعٍ٘ح عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪ ، ٛ‬أٍا تاىْغثح ىرأش‪ٞ‬ش األٍٖاخ‬
‫اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو ‪ ،‬فقذ ماُ عاى‪ ٜ‬اىَعْ٘‪ٝ‬ح ف‪ ٜ‬حاىح اىرأش‪ٞ‬ش عي‪ّ ٚ‬غثح‬
‫اىقث٘ه ٗغ‪ٞ‬ش ٍعْ٘‪ ٛ‬ف‪ ٜ‬حاىح اىرأش‪ٞ‬ش عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ‪0‬‬
‫اذعح ٍِ ّرائط غ٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍخريفح األصو‬
‫اى٘ساش‪ ٜ‬خاله اىفرشج ٍِ ‪ 2008 /7/4‬إى‪ 2008 /7/19 ٚ‬أُ أقو ّغثح قث٘ه ٗ أقو مَ‪ٞ‬ح غزاء‬
‫ٍين‪ ٜ‬ماّد ف‪ ٜ‬اىغُذاتح األٗى‪ ٚ‬ف‪ ٜ‬ح‪ ِٞ‬أُ اىغُذاتح اىصاّ‪ٞ‬ح ععيد أعي‪ّ ٚ‬غثح قث٘ه ٗمَ‪ٞ‬ح غزاء‬
‫اىغزاء اىَيل‪ ٛ‬اى َُْرعح ‪0‬‬ ‫ٍين‪ٗ ،ٜ‬احريد اىغُذاتح اىصاىصح ٍشمضاً ٗعط‪ٞ‬ا ً ف‪ّ ٜ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح‬
‫ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف ‪ %48,1‬ت‪َْٞ‬ا ماُ ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‬
‫‪ 1,141‬ظٌ ‪ 0‬ماُ ذأش‪ٞ‬ش مو ٍِ ّغثح قث٘ه اى‪ٞ‬شقاخ اىَطعٍ٘ح ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‬
‫ٍعْ٘‪ ،ٛ‬أ‪ٝ‬عا ً ماُ ذأش‪ٜ‬س مو ٍِ األٍٖاخ اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ‬
‫ج‬
‫ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪0 ٛ‬‬
‫ف‪ ٜ‬غ٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍخريفح األصو اى٘ساش‪ ٜ‬خاله‬
‫اىفرشج ٍِ ‪ 2008 /8/6‬إى‪ ،2008 /8/15 ٚ‬أعطد اىغُذاتح األٗى‪ ٚ‬أقو ّغثح قث٘ه ٗمَ‪ٞ‬ح ٍِ‬
‫ف‪ ٜ‬ح‪ِٞ‬‬ ‫اىغزاء اىَين‪ ٜ‬ت‪َْٞ‬ا ععيد اىغُذاتح اىصاّ‪ٞ‬ح أعي‪ّ ٚ‬غثح قث٘ه ٗمَ‪ٞ‬ح ٍِ اىغزاء اىَين‪،ٜ‬‬
‫احريد اىغُذاتح اىصاىصح ٍشمضاً ٗعط‪ٞ‬ا ً ف‪ّ ٜ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاءاىَين‪ ٜ‬اى َُْرعح ‪ 0‬ماُ ٍر٘عػ‬
‫ّغثح اىقث٘ه ٗ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ىٖزٓ اىط٘ائف ‪ 0,820 ٗ %57,7‬ظٌ‪ ،‬عي‪ٚ‬‬
‫اىر٘اى‪ 0ٜ‬ماّد ّغثح قث٘ه اى‪ٞ‬شقاخ اىَطعٍ٘ح عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪ ٛ‬ج ‪،‬‬
‫ٍِ ّغثح‬ ‫ت‪َْٞ‬ا ماُ ذأش‪ٞ‬ش مو ٍِ األٍٖاخ اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو‬

‫‪2‬‬
‫اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪ ٛ‬ف‪ ٜ‬حاىح اىرأش‪ٞ‬ش عي‪ّ ٚ‬غثح اىقث٘ه ٍٗعْ٘‪ ٛ‬ف‪ٜ‬‬
‫حاىح اىرأش‪ٞ‬ش عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ‪0‬‬
‫ٗف‪َٞ‬ا ‪ٝ‬رعيق بغ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍخريفح‬
‫األصو اى٘ساش‪ ٜ‬خاله اىفرشج ٍِ ‪ 2009 /3/22‬إى‪ ،2009 /4/6 ٚ‬أظٖشخ اىْرائط أُ اىغُذاتح‬
‫األٗى‪ ٚ‬أعطد أعي‪ّ ٚ‬غثح قث٘ه ٗأّرعد أمثش مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ ٜ‬ذوخٕا اىغُذاتح اىصاّ‪ٞ‬ح‪ .‬ف‪ٜ‬‬
‫ح‪ ِٞ‬أعطد اىغُذاتح اىصاىصح أقو ّغثح هىقث٘ه ٗ أقو مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ 0 ٜ‬ماّد ٍر٘عطاخ‬
‫ٕزٓ اىط٘ائف ‪ّ ٪ 35,4‬غثح اىقث٘ه ٗ ‪ 0,519‬ظٌ غزاء ٍين‪ 0 ٜ‬ماُ ذأش‪ٞ‬ش ّغثح قث٘ه اى‪ٞ‬شقاخ‬
‫اىَطعٍ٘ح عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح عاى‪ ٜ‬اىَعْ٘‪ٝ‬ح‪ ،‬مزىل ماُ ذأش‪ٞ‬ش مو ٍِ األٍٖاخ‬
‫اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‬
‫عاى‪ ٜ‬اىَعْ٘‪ٝ‬ح‪0‬‬
‫ٗتصفح عاٍح‪ ،‬فقذ أٗظحد ّرائط اىط٘ائف اىَطعٍ٘ح ت‪ٞ‬شقاخ راخ أصو ٗساش‪ٜ‬‬
‫ٍخريف عِ األصو اى٘ساش‪ ٜ‬ىطائفح االّراض اىَعشٗظح عاتقا ً ٍا ‪ٝ‬ي‪:ٜ‬‬
‫ف‪ ٜ‬حاىح اىغُذاتاخ اىصالز اىخاصح تإغاس اإلّراض‪ ،‬ععيد اىغُذاتح اىصاّ‪ٞ‬ح أعي‪ّ ٚ‬غثح‬
‫‪ٗ ،)%42,8‬احريد‬ ‫قث٘ه ( ‪ )%55,2‬ف‪ ٜ‬ح‪ ِٞ‬أُ أقو ّغثح قث٘ه ٗظذخ ف‪ ٜ‬اىغُذاتح اىصاىصح (‬
‫اىغُذاتح األٗى‪ ٚ‬اىَشمض اىصاّ‪ ٜ‬ف‪ّ ٜ‬غثح اىقث٘ه ( ‪0)%50,4‬‬
‫ف‪ ٜ‬حاىح مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرط‪ ،‬فقذ أّرعد اىغُذاتح األٗى‪ ٚ‬أمثش مَ‪ٞ‬ح ذوخٕا اىغُذاتح‬
‫اىصاّ‪ٞ‬ح شٌ اىغُذاتح اىصاىصح ( ‪ 0,841‬ظٌ‪ 0,836 ،‬ظٌ ٗ ‪0,701‬ظٌ عي‪ ٚ‬اىرشذ‪ٞ‬ة) ‪0‬‬
‫ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف ‪ %49,6‬ت‪َْٞ‬ا ماُ ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ٜ‬‬
‫اى َُْرعح ‪0,804‬ظٌ ‪0‬‬
‫أٗظحد ٍعظٌ اىط٘ائف اى َُخرثشج ٗظ٘د عالقاخ غ‪ٞ‬ش ٍعْ٘‪ٝ‬ح ت‪ّ ِٞ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح‬
‫اىغزاء اىَين‪ ٜ‬اى َُْرط‪ ،‬مزىل ماُ ذأش‪ٞ‬ش مو ٍِ أألٍٖاخ اىنارتح ٗعذد األقشاص اىَغطاج تاىْحو‬
‫عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرط غ‪ٞ‬ش ٍعْ٘‪ ،ٛ‬ف‪َٞ‬ا عذا ٍعَ٘عح اىط٘ائف‬
‫خاله اىفرشج ٍات‪ 2009 /3/22 ِٞ‬إى‪ٚ‬‬ ‫اىخاٍغح (عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ )ٜ‬اىر‪ ٜ‬اُخرثشخ‬
‫‪ 2009 /4/6‬ح‪ٞ‬س أظٖشخ عالقاخ عاى‪ٞ‬ح اىَعْ٘‪ٝ‬ح ‪0‬‬

‫‪3‬‬
‫ب‪ .‬طىائف تم تطعيمها تيزقات من نفس األصل الىراثي لها‬
‫أظش‪ٝ‬د ٕزٓ اىذساعح تاعرخذاً غ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ،ٜ‬اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ٜ‬‬
‫ٗاىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬عذ‪َٝ‬ح اىَيناخ اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪0 ٜ‬‬
‫ف‪َٞ‬ا ‪ٝ‬رعيق بغ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ‬
‫األصو اى٘ساش‪ ٜ‬خاله اىفرشج ٍِ ‪ 2009 /4/8‬إى‪ ،2009 /4/20 ٚ‬أٗظحد اىْرائط أُ اىغُذاتح‬
‫األٗى‪ ٚ‬أّرعد أقو ّغثح قث٘ه ٗأقو مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ ،ٜ‬ف‪ّ ٜ‬فظ اى٘قد أّرعد اىغُذاتح‬
‫اىصاىصح أعي‪ّ ٚ‬غثح قث٘ه ٗمَ‪ٞ‬ح غزاء ٍين‪ ،ٜ‬أٍا تاىْغثح ىيغُذاتح اىصاّ‪ٞ‬ح فقذ احريد اىَشمض اىصاّ‪ٜ‬‬
‫ف‪ّ ٜ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ 0 ٜ‬ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف ‪ %32,5‬ت‪َْٞ‬ا‬
‫ماُ ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ‪0,978‬ظٌ‪ 0‬ماُ ذأش‪ٞ‬ش ّغثح قث٘ه اى‪ٞ‬شقاخ اىَطعٍ٘ح‬
‫عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ٍعْ٘‪ٝ‬ا ً‪ ،‬ف‪ ٜ‬ح‪ ِٞ‬ماُ ذأش‪ٞ‬ش مو ٍِ األٍٖاخ اىنارتح ٗعذد‬
‫األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪0 ٛ‬‬
‫ف‪ ٜ‬حاىح غ٘ائف اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ٜ‬‬
‫خاله اىفرشج ٍِ ‪ 2009 /7/15‬إى‪ ،2009 /8/2 ٚ‬عُعيد أعي‪ّ ٚ‬غثح ىيقث٘ه ٗأمثش مَ‪ٞ‬ح ٍِ‬
‫اىغزاء اىَين‪ ٜ‬ف‪ ٜ‬اىغُذاتح اىصاىصح‪ ،‬ف‪ ٜ‬ح‪ ِٞ‬أّرعد اىغُذاتح األٗى‪ ٚ‬أقو ّغثح قث٘ه ٗأقو مَ‪ٞ‬ح‬
‫ٍِ اىغزاء اىَين‪ ، ٜ‬أٍا تاىْغثح ىيغُذاتح اىصاّ‪ٞ‬ح فقذ احريد اىَشمض اىصاّ‪ ٜ‬ف‪ّ ٜ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح‬
‫اىغزاء اىَين‪ ٜ‬اى َُْرط ‪ٗ 0‬قذ أعطد ٕزٓ اىط٘ائف ‪ ٪ 64,6‬ف‪ٍ ٜ‬ر٘عػ ّغثح اىقث٘ه ٗ ‪0,959‬‬
‫ظٌ ف‪ٍ ٜ‬ر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ‪ 0‬ماُ ذأش‪ٞ‬ش ّغثح قث٘ه اى‪ٞ‬شقاخ اىَطعٍ٘ح عي‪ٚ‬‬
‫مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪ٗ ،ٛ‬مزىل فقذ ماُ ذأش‪ٞ‬ش مو ٍِ األٍٖاخ اىنارتح ٗعذد‬
‫األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪.ٛ‬‬
‫ٗأظٖشخ ّرائط غ٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو‬
‫اى٘ساش‪ ٜ‬خاله اىفرشج ٍِ ‪ 2009 /8/13‬إى‪2009 /8/28 ٚ‬أُ اىغُذاتح األٗى‪ ٚ‬أّرعد أقو ّغثح‬
‫قث٘ه ٗأدّ‪ ٚ‬مَ‪ٞ‬ح ٍِ اىغزاء اىَين‪ ،ٜ‬ت‪َْٞ‬ا أّرعد اىغُذاتح اىصاىصح أمثش ّغثح اىقث٘ه ٗمَ‪ٞ‬ح‬
‫اىغزاءاىَين‪ ،ٜ‬عي‪ ٚ‬اىْق‪ٞ‬ط ٍِ رىل فقذ احريد اىغُذاتح اىصاّ‪ٞ‬ح اىَشمض اىصاّ‪ ٜ‬ف‪ّ ٜ‬غثح اىقث٘ه‬
‫‪ ،%70,2‬ت‪َْٞ‬ا ماُ‬ ‫ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرط ‪ 0‬ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف‬
‫‪ 1,535‬ظٌ ‪ 0‬ذٌ اىحص٘ه عي‪ ٚ‬دسظح عاى‪ٞ‬ح ٍِ‬ ‫ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح ٕ‪ٜ‬‬
‫اىَعْ٘‪ٝ‬ح عْذ دساعح ذأش‪ٞ‬ش ّغثح قث٘ه اى‪ٞ‬شقاخ اىَطعٍ٘ح عي‪ ٚ‬مَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح‪ ،‬ف‪ٜ‬‬

‫‪4‬‬
‫ح‪ ِٞ‬ماُ ذأش‪ٞ‬ش مو ٍِ األٍٖاخ اىنارتح ٗعذد األقشاص اى َُغطاج تاىْحو عي‪ ٚ‬مو ٍِ ّغثح‬
‫اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرعح غ‪ٞ‬ش ٍعْ٘‪0 ٛ‬‬
‫أٗظحد اىْرائط اىغاتقح اىخاصح تاىط٘ائف اىَطعٍ٘ح ت‪ٞ‬شقاخ ٍِ ّفظ األصو‬
‫‪ٗ %65,6‬‬ ‫اى٘ساش‪ ٜ‬أُ اىغُذاتح اىصاىصح أعطد أعي‪ٍ ٚ‬ر٘عػ ّغثح قث٘ه ٗمَ‪ٞ‬ح غزاء ٍين‪( ٜ‬‬
‫‪ 1,333‬ظٌ عي‪ ٜ‬اىر٘اى‪ ،)ٜ‬ف‪ ٜ‬ح‪ ِٞ‬ماُ أقو ٍر٘عػ ىْغثح اىقث٘ه ٗىنَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬ف‪ٜ‬‬
‫اىغُذاتح األٗى‪ 0,905 ٗ %44,8 ( ٚ‬ظٌ عي‪ ٜ‬اىرشذ‪ٞ‬ة)‪ٗ ،‬احريد اىغُذاتح اىصاّ‪ٞ‬ح اىَشمض اىصاّ‪ٜ‬‬
‫ف‪ّ ٜ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ 0,905 ٗ %56,8 ( ٜ‬ظٌ عي‪ ٜ‬اىرشذ‪ٞ‬ة) ‪0‬‬
‫ماُ ٍر٘عػ ّغثح اىقث٘ه ىٖزٓ اىط٘ائف ‪ %55,8‬ت‪َْٞ‬ا ماُ ٍر٘عػ مَ‪ٞ‬ح اىغزاء اىَين‪ٜ‬‬
‫اى َُْرعح ٕ‪1,157 ٜ‬ظٌ‪0‬‬
‫أٗظحد ظَ‪ٞ‬ع اىط٘ائف اى َُخرثشج ٗظ٘د عالقاخ غ‪ٞ‬ش ٍعْ٘‪ٝ‬ح ت‪ّ ِٞ‬غثح اىقث٘ه ٗمَ‪ٞ‬ح‬
‫اىغزاء اىَين‪ ٜ‬اى َُْرط ‪ٗ ،‬مزىل ت‪ ِٞ‬ذأش‪ٞ‬ش مو ٍِ األٍٖاخ اىنارتح ٗعذد األقشاص اىَغطاج تاىْحو‬
‫عي‪ ٚ‬مو ٍِ ّغثح اىقث٘ه ٗمَ‪ٞ‬ح اىغزاء اىَين‪ ٜ‬اى َُْرط ‪0‬‬
‫في ضوء النتائج السابقة والتي تتعلق بتقييم بعض العوامل المؤثرة على إنتاج الغذاء‬
‫الملكي بطوائف نحل العسل ‪ ،‬أعطت الطوائف المطعومة بيرقات من نفس األصل الوراثي‬
‫نسبة قبول أكبر (‪ )%55,8‬وكمية غذاء ملكي أعلى ( ‪ 1,157‬جم ) من نظيرتها المطعومة‬
‫بيرقات ذات أصل وراثي مختلف عن األصل الوراثي لطائفة االنتاج ( ‪ %49,6‬و ‪0,804‬‬
‫جم‪ ،‬على التوالي)‪0‬‬
‫‪ .2‬تحذيذ نسة األحماض األمينية والذهنية في عينات الغذاء الملكي ال ُمنتجة‬
‫أ ‪ 0‬تحذيذ نسة األحماض األمينية‬
‫أظٖشخ اىْرائط اىخاصح ب غ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا‬
‫ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ ٜ‬عْذٍا ماُ ٍر٘عػ ّغثح ظٖ٘س األٍٖاخ اىنارتح ‪ ،%71‬أُ‬
‫ٍر٘عطاخ ّغة األحَاض األٍ‪ْٞٞ‬ح األعاع‪ٞ‬ح ماّد ‪ ٪ 0,569‬ىيصش‪ ٪ 0,740 ، ِّٞ٘ٝ‬هىفاى‪، ِٞ‬‬
‫‪ ٪ 0,116‬ىيَ‪ٞ‬ص‪ ٪ 0,507 ، ِّٞ٘ٞ‬ىأل‪ٛ‬صٗى٘ع‪ ٪ 1,023 ، ِٞ‬ىي‪٘ٞ‬ع‪ ٪ 0,402 ، ِٞ‬هىف‪ْٞٞ‬و‬
‫آالّ‪ ٪ 0,382 ، ِٞ‬هىٖغرذ‪ ٪ 0،569 ٗ ِٝ‬هى‪ٞ‬غ‪ 0ِٞ‬ف‪ ٜ‬حاىح األحَاض األٍ‪ْٞٞ‬ح غ‪ٞ‬ش األعاع‪ٞ‬ح‪،‬‬
‫ماّد اهٍر٘عطاخ ماألذ‪، ٪ 0,949 ، ٪ 0,015 ، ٪ 1,068 ، ٪ 1,007 ، ٪ 2,230 ٜ‬‬

‫‪5‬‬
‫ىألعثاسذ‪ٞ‬ل ‪ ،‬اهع‪ٞ‬ش‪ ، ِٝ‬اه ضى٘ذاٍ‪ٞ‬ل ‪ ،‬اهتشٗى‪، ِٞ‬‬ ‫‪٪ 0،480 0,441‬‬ ‫‪ٗ ، ٪ 0,676‬‬
‫اهظي‪ٜ‬ط‪ ، ِٝ‬أالأالّ‪ ، ِٞ‬اىر‪ٞ‬شٗع‪ٗ ِٞ‬األسظ‪ ، ِْٞٞ‬عي‪ ٚ‬اىر٘اى‪0 ٜ‬‬
‫عْذٍا ععيد ّغثح ظٖ٘س األٍٖاخ اىنارتح ‪ %80‬ف‪ ٜ‬غ٘ائف اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ ٜ‬اىر‪ ٜ‬ذٌ‬
‫ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ ، ٜ‬ماّد ٍر٘عطاخ ّغة األحَاض األٍ‪ْٞٞ‬ح األعاع‪ٞ‬ح‬
‫‪ ،%0,111‬األ‪ٝ‬ضٗى‪ٞ‬غ‪ِٞ‬‬ ‫‪ ،%0,672‬اىَ‪ٞ‬ص‪ِّٞ٘ٞ‬‬ ‫ٍا ‪ٝ‬ي‪ :ٜ‬اىصش‪ ،%0,509 ِّٞ٘ٝ‬اىفاى‪ِٞ‬‬
‫‪ ،%0,464‬اىي‪٘ٞ‬ع‪ ،%0,901 ِٞ‬اىف‪ْٞٞ‬و آالّ‪ ،%0,338 ِٞ‬اىٖغرذ‪ٗ %0,313 ِٝ‬اىي‪ٞ‬غ‪ِٞ‬‬
‫‪ 0%0,763‬ت‪َْٞ‬ا ماّد ٍر٘عطاخ ّغة األحَاض األٍ‪ْٞٞ‬ح غ‪ٞ‬ش األعاع‪ٞ‬ح ماألذ‪،%2,285 :ٜ‬‬
‫ٗ ‪%0,413‬‬ ‫‪%0,359 ،%0,586 ،%0,830 ،%0,012 ،%0,923 ،%0,889‬‬
‫هألعثاس ذ‪ٜ‬ك‪ ،‬اىغ‪ٞ‬ش‪ ،ِٞٝ‬اىعي٘ذاٍ‪ٞ‬ل‪ ،‬اىثشٗى‪ ،ِٞ‬اىعي‪ٞ‬غ‪ ، ِٞ‬ا‪ٟ‬الّ‪ ،ِٞ‬اىر‪ٞ‬شٗع‪ٗ ،ِٞ‬األسظ‪ِْٞٞ‬‬
‫عي‪ ٚ‬اىر٘اى‪0 ٜ‬‬
‫ف‪َٞ‬ا ‪ٝ‬رعيق تط٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو‬
‫اى٘ساش‪ ،ٜ‬فقذ أعطد األحَاض األٍ‪ْٞٞ‬ح األعاع‪ٞ‬ح – عْذٍا ماّد ّغثح ظٖ٘س األٍٖاخ اىنارتح‬
‫‪ - %80‬اىَر٘عطاخ اىراى‪ٞ‬ح ‪ :‬اىصش‪ ،%0,502 ِّٞ٘ٝ‬اىفاى‪ ،%0,660 ِٞ‬اىَ‪ٞ‬ص‪،%0,067 ِّٞ٘ٞ‬‬
‫األ‪ٝ‬ضٗى‪ٞ‬غ‪ ،%0,451 ِٞ‬اىي‪٘ٞ‬ع‪ ،%0,869 ِٞ‬اىف‪ْٞٞ‬و آالّ‪ ،%0,325 ِٞ‬اىٖغرذ‪%0,301 ِٝ‬‬
‫‪،%2,259‬‬ ‫األٍ‪ْٞٞ‬ح غ‪ٞ‬ش األعاع‪ٞ‬ح‬ ‫ٗ اىي‪ٞ‬غ‪ٗ 0%0,895 ِٞ‬ماّد ٍر٘عطاخ األحَاض‬
‫‪ %0,393 ٗ %0,356 ،%0,602 ،%0,819 ،%0,013 ،%0,977 ،%0,918‬ىنو‬
‫ٍِ األعثاساذل‪ ،‬اىغ‪ٞ‬ش‪ ، ِٞٝ‬اىعي٘ذاٍ‪ٞ‬ل ‪ ،‬اىثشٗى‪ ، ِٞ‬اىعي‪ٞ‬غ‪ ، ِٞ‬ا‪ٟ‬الّ‪ ، ِٞ‬اىر‪ٞ‬شٗع‪ٗ ، ِٞ‬األسظ‪ِْٞٞ‬‬
‫عي‪ ٚ‬اىر٘اى‪0 ٜ‬‬
‫ماّد أعي‪ّ ٚ‬غة‬ ‫ٗاىخاصح تْغة األحَاض األٍ‪ْٞٞ‬ح‪ ،‬فقذ‬ ‫ذثعا ُ ىيْرائط اىغاتقح‬
‫ذالٕا اىٖع‪ِٞ‬‬ ‫هألحَاض األٍ‪ْٞٞ‬ح األعاع‪ٞ‬ح ٍ٘ظ٘دج ف‪ ٜ‬ع‪ْٞ‬اخ غ٘ائف عالىح اىَصش‪ ٛ‬اىْق‪ٜ‬‬
‫اإل‪ٝ‬طاى‪ ٜ‬شٌ اىنشّ‪٘ٞ‬ى‪ ، ٜ‬ف‪َٞ‬ا عذا حاٍط اىي‪ٞ‬غ‪ ِٞ‬اىز‪ ٛ‬أعط‪ ٚ‬أعي‪ّ ٚ‬غثح ىٔ ف‪ ٜ‬ع‪ْٞ‬اخ غ٘ائف‬
‫شٌ ع‪ْٞ‬اخ غ٘ائف عالىح اىْحو‬ ‫اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬ذالٕا ع‪ْٞ‬اخ غ٘ائف اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ٜ‬‬
‫اىَصش‪ ٛ‬اىْق‪ 0 ٜ‬أٍا تاىْغثح ىألحَاض األٍ‪ْٞٞ‬ح غ‪ٞ‬ش األعاع‪ٞ‬ح ‪ ،‬فيقذ ععيد أعي‪ّ ٚ‬غة ىٖا ف‪ٜ‬‬
‫ع‪ْٞ‬اخ غ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ، ٜ‬ت‪َْٞ‬ا ذأسظح اىَشمض اُ اىصاّ‪ٗ ٜ‬اىصاىس ت‪ ِٞ‬مو‬
‫ٍِ اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ٗ ٜ‬اىنشّ‪٘ٞ‬ى‪0 ٜ‬‬

‫‪6‬‬
‫ب ‪ 0‬تحذيذ نسة األحماض الذهنية‬
‫ماّد ّغة ٍر٘عطاخ األحَاض اىذْٕ‪ٞ‬ح ف‪ ٜ‬ع‪ْٞ‬اخ اىغزاء اىَين‪ ٜ‬اىَخريفح اىخاصح‬
‫بغ٘ائف عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ٜ‬‬
‫ما‪ٟ‬ذ‪ :ٜ‬حاٍط اىٖنغاد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,113‬حاٍط األٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,024‬حاٍط‬
‫‪ ،%0,301‬حاٍط‬ ‫اىغ‪ٞ‬ض‪ ،‬ع‪ٞ‬ض‪-9,12 -‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,426‬حاٍط األ‪ٝ‬ن٘عاّ٘‪ٝ‬ل‬
‫ذ‪ٞ‬رشام٘عاّ٘‪ٝ‬ل ‪ٗ %0,159‬حاٍط اىغ‪ٞ‬ض‪-9 -‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ 0%0,162‬أٍا تاىْغثح ىحاٍط‬
‫‪ٕٞ -10‬ذسٗمغ‪ -2 ٜ‬د‪ٝ‬غ‪ْٝ٘ٞ‬ل فقذ ماّد ّغثرٔ ‪0%2,731‬‬
‫ٗذثعا ً ىْرائط غ٘ائف اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو‬
‫ّغة األحَاض اىذْٕ‪ٞ‬ح ف‪ ٜ‬ع‪ْٞ‬اخ اىغزاء اىَين‪ ٜ‬اىَخريفح ما‪ٟ‬ذ‪:ٜ‬‬ ‫اى٘ساش‪ ،ٜ‬فقذ ماّد‬
‫‪ %2,773‬ههحاٍط ‪ٕٞ -10‬ذسٗمغ‪ -2 ٜ‬د‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ %0,113 ،‬ىوحاٍط اىٖنغاد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪،‬‬
‫‪ %0,373‬ىيحاٍط‬ ‫‪ %0,049‬ىوحاٍط األٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ %0,119 ،‬ىوحاٍط ع‪ٞ‬ض‪،‬‬
‫ع‪ٞ‬ض ‪ 9,12‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪،‬‬
‫اىخاصح‬ ‫أظٖشخ ّرائط ّغة األحَاض اىذْٕ‪ٞ‬ح ف‪ ٜ‬ع‪ْٞ‬اخ اىغزاء اىَين‪ ٜ‬اىَخريفح‬
‫أُ ٍر٘عطاذٖا‬ ‫بغ٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬اىر‪ ٜ‬ذٌ ذطع‪َٖٞ‬ا ت‪ٞ‬شقاخ ٍِ ّفظ األصو اى٘ساش‪ٜ‬‬
‫ما‪ٟ‬ذ‪ :ٜ‬حاٍط اىٖنغاد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,173‬حاٍط األٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,089‬حاٍط‬
‫‪ ،%0,337‬حاٍط‬ ‫اىغ‪ٞ‬ض‪ ،‬ع‪ٞ‬ض‪-9,12 -‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ ،%0,223‬حاٍط األ‪ٝ‬ن٘عاّ٘‪ٝ‬ل‬
‫ذ‪ٞ‬رشام٘عاّ٘‪ٝ‬ل ‪ٗ %0,162‬حاٍط اىغ‪ٞ‬ض‪-9 -‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ .%0,405‬أٍا تاىْغثح ىحاٍط‬
‫‪ٕٞ -10‬ذسٗمغ‪ -2 ٜ‬د‪ٝ‬غ‪ْٝ٘ٞ‬ل فقذ ماّد ّغثرٔ ‪0%2,033‬‬
‫ٗغثقا ً ىْرائط األحَاض اىذْٕ‪ٞ‬ح‪ ،‬فقذ اذعح أُ أعي‪ّ ٚ‬غة ٍِ ذيل األحَاض قذ عُعو‬
‫ف‪ ٜ‬ع‪ْٞ‬اخ اىغزاء اىَين‪ ٜ‬ىط٘ائف اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ ٜ‬ف‪ ٜ‬ح‪ ِٞ‬ذأسظح اىَشمض اُ اىصاّ‪ٗ ٜ‬اىصاىس‬
‫‪ ،‬تاعرصْاء مو ٍِ حاٍع‪ٜ‬‬ ‫ف‪َٞ‬ا ت‪ ِٞ‬ع‪ْٞ‬اخ اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ ٗ ٜ‬عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ٜ‬‬
‫‪ٕٞ -10‬ذسٗمغ‪ -2 - ٜ‬د‪ٝ‬غ‪ْٝ٘ٞ‬ل ٗ اىغ‪ٞ‬ض‪ ،‬ع‪ٞ‬ض‪-9,12 -‬أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل ‪ 0‬فف‪ ٜ‬حاىح اهحاٍط‬
‫ذيرٖا‬ ‫‪ٕٞ -10‬ذسٗمغ‪ -2 - ٜ‬د‪ٝ‬غ‪ْٝ٘ٞ‬ل فقذ ععيد أعي‪ّ ٚ‬غثح ىٔ ف‪ ٜ‬ع‪ْٞ‬اخ اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪ٜ‬‬
‫عالىح اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬شٌ اىٖع‪ ِٞ‬اىنشّ‪٘ٞ‬ى‪ 0 ٜ‬أٍا تاىْغثح ىوحاٍط اىغ‪ٞ‬ض‪ ،‬ع‪ٞ‬ض‪-9,12 -‬‬
‫أٗمراد‪ٝ‬غ‪ْٝ٘ٞ‬ل فقذ ععوخ أعي‪ّ ٚ‬غثح ىٔ ف‪ ٜ‬ع‪ْٞ‬اخ اىْحو اىَصش‪ ٛ‬اىْق‪ ٜ‬ذثع خٕا ع‪ْٞ‬اخ اىٖع‪ِٞ‬‬
‫اىنشّ‪٘ٞ‬ى‪ ٜ‬شٌ اىٖع‪ ِٞ‬اإل‪ٝ‬طاى‪0 ٜ‬‬

‫‪7‬‬
‫دساصاخ عهى إَراج انغزاء انًهكي في طىائف َذم انعضم‬

‫سصانح يقذيح يٍ‬

‫داذى أدًذ ششف انذيٍ‬


‫تكانىسيىس في انعهىو انزساعيح (وقايح َثاخ)‪ -‬كهيح انزساعح ‪ -‬جايعح انقاهشج‪2006 ،‬‬

‫نهذصىل عهى دسجح‬

‫انًاجضريش‬

‫في‬

‫انعهىو انزساعيح‬
‫(دششاخ إقرصاديح )‬

‫قشـــــــو انذششاخ اإلقرصاديح وانًثيذاخ‬


‫كهيــــــح انزساعـــــح‬
‫جـايعــح انقــاهشج‬
‫يصـــــــــس‬

‫‪2010‬‬
‫دساصاخ عهى إَراج انغزاء انًهكي في طىائف َذم انعضم‬

‫سصانح ياجضريش‬
‫في انعهىو انزساعيح‬
‫(دششاخ إقرصاديح)‬

‫يقذيح يٍ‬

‫داذى أدًذ ششف انذيٍ‬


‫تكانىسيىس في انعهىو انزساعيح (وقايح َثاخ)‪ -‬كهيح انزساعح ‪ -‬جايعح انقاهشج‪2006 ،‬‬

‫نعُح انؽكى‬

‫‪........................................................ ..................................‬‬ ‫د‪ .‬يذًذ تهجد شاوس‬


‫أصرار انذششاخ اإلقرصاديح – كهيح انزساعح – جايعح كفش انشيخ )‪(12B‬‬

‫‪.................................................................................................‬‬ ‫د‪ .‬يذًذ عطيح عىيش‬


‫أصرار انذششاخ اإلقرصاديح – كهيح انزساعح – جايعح انقاهشج‬

‫د‪ .‬يذًذ عثذ انىهاب عثذ‬


‫انفراح‪..............................................................................‬‬
‫أصرار انذششاخ اإلقرصاديح – كهيح انزساعح – جايعح انقاهشج‬

‫د‪ .‬يذًىد انضيذ انضيذ َىس ‪...................................................................................‬‬


‫أصرار انذششاخ اإلقرصاديح – كهيح انزساعح – جايعح انقاهشج‬

‫‪2010 /‬‬ ‫‪/‬‬ ‫التاريخ‪:‬‬


‫دساصاخ عهى إَراج انغزاء انًهكي في طىائف َذم انعضم‬

‫سصانح ياجضريش‬
‫في انعهىو انزساعيح‬
‫(دششاخ إقرصاديح)‬

‫يقذيح يٍ‬

‫داذى أدًذ ششف انذيٍ‬


‫تكانىسيىس في انعهىو انزساعيح (وقايح َثاخ)‪ -‬كهيح انزساعح ‪ -‬جايعح انقاهشج‪2006 ،‬‬

‫نعُح اإلشزاف‬

‫انذكرىس‪ /‬يذًىد انضيذ انضيذ َىس‬


‫أصرار انذششاخ اإلقرصاديح‪ -‬كهيح انزساعح ‪ -‬جايعح انقاهشج‬

‫انذكرىس‪ /‬يذًذ عثذ انىهاب عثذ انفراح‬


‫أصرار انذششاخ اإلقرصاديح – كهيح انزساعح ‪ -‬جايعح انقاهشج‬
‫انذسجح‪ :‬انًاظسرُز‬ ‫اصى انطانة‪ :‬ؼاذى أؼًذ شزف انذٍَ‬
‫عُىاٌ انشصانح‪ :‬دراساخ عهً إَراض انغذاء انًهكٍ فٍ غىائف َؽم انعسم‬
‫انًششفىٌ ‪ :‬دكرىس ‪ :‬يؽًىد انسُذ َىر‬
‫دكرىس ‪ :‬يؽًذ عثذ انىهاب عثذ انفراغ‬

‫‪2010 /‬‬ ‫‪/‬‬ ‫قضى‪ :‬انؽشزاخ اإلقرصادَح وانًثُذاخ فشع‪ :‬انؽشزاخ ذاسيخ يُخ انذسجح‪:‬‬
‫انًضرخهص انعشتي‬
‫– كهُح‬ ‫ذى اظزاء هذا انثؽس فٍ وؼذج تؽىز انُؽم تًؽطح انرعارب انزراعُح‬
‫انزراعح‪-‬ظايعح انقاهزج تانعُزج ؼُس كاٌ انهذف انزئُسٍ نهذا انثؽس هى ذقُُى تعط‬
‫انعىايم انًؤشزج عهً اَراض انغذاء انًهكٍ فٍ غىائف َؽم انعسم و ذؽذَذ َسة األؼًاض‬
‫األيُُُح وانذهُُح فٍ عُُاخ انغذاء انًهكٍ ان ًُُرعح‪0‬‬
‫ذى ذقسُى انرعارب انخاصح تانهذف األول إنً غىائف ذى ذطعًُها تُزقاخ يٍ أصم‬
‫وراشٍ يخرهف عٍ أصهها انىراشٍ و غىائف ذى ذطعًُها تُزقاخ يٍ َفس األصم انىراشٍ‬
‫نها‪0‬‬
‫ذى إظزاء ذعارب انطىائف انرٍ ذى ذطعًُها تُزقاخ يٍ أصم وراشٍ يخرهف عٍ‬
‫أصهها انىراشٍ تاسرعًال كم يٍ سالنح انُؽم انًصزٌ انُقٍ وانهعٍُ انكزَُىنٍ ‪ 0‬ونقذ‬
‫أظهزخ َرائط غىائف سالنح انُؽم انًصزٌ انُقٍ ؼذوز اَخفاض يهؽىظ فٍ َسثح قثىل‬
‫انُزقاخ انًطعىيح وكًُح انغذاء انًهكٍ ان ًُُرط عُذ يقارَرها تانهعٍُ انكزَُىنٍ‪0‬‬
‫فقذ أظزَد‬‫أيا ذعارب الغىائف انرٍ ذى ذطعًُها تُزقاخ يٍ َفس األصم انىراشٍ نها ل‬
‫تاسرخذاو سالنح انُؽم انًصزٌ انُقٍ وانهعٍُ االَطانٍ وانكزَُىنٍ ‪ 0‬ونقذ أوظؽد‬
‫انُرائط إٌ انهعٍُ انكزَُىنٍ هى األفعم اسرخذيا ً إلَراض انغذاء انًهكٍ فقذ أعطً أكثز‬
‫َسثح قثىل وأكثز كًُح يٍ انغذاء انًهكٍ عُذ يقارَره تثاقٍ انهعٍ وانسالالخ انًسرخذيح ‪0‬‬
‫انُؽم انًصزٌ انُقٍ فٍ ؼٍُ‬ ‫كًا سعم انهعٍُ اإلَطانٍ َسثح قثىل أعهً يٍ سالنح‬
‫أعطد األخُزج كًُح غذاء يهكٍ يساوَح ذقزَثا ً نهكًُح انرٍ أعطاها انهعٍُ اإلَطانٍ تزَادج‬
‫ؼىانٍ ‪ 0.02‬ظى أكثز نصانػ سالنح انُؽم انًصزٌ انُقٍ‪0‬‬
‫تانُسثح نرؽذَذ َسة األؼًاض األيُُُح وانذهُُح‪ ،‬فهقذ أشارخ انُرائط إنً أٌ َسة‬
‫األؼًاض األيُُُح األساسُح كاَد أكثز يا ًَكٍ فٍ عُُاخ غىائف سالنح انُؽم انًصزٌ‬
‫انُقٍ ذالها انهعٍُ اإلَطانٍ شى انكزَُىنٍ فًُا عذا ؼايط انهُسٍُ انذي سُعهد اعهً َسثح‬
‫نه فٍ عُُاخ غىائف انهعٍُ انكزَُىنٍ شى عُُاخ غىائف انهعٍُ اإلَطانٍ وأخُزاً عُُاخ‬
‫غىائف سالنح انُؽم انًصزٌ انُقٍ ‪ 0‬ونقذ وظذخ أعهً َسة يٍ األؼًاض األيُُُح غُز‬
‫األساسُح فٍ عُُاخ غىائف سالنح انُؽم انًصزٌ انُقٍ أيا انًزكز انصاٍَ وانصانس فقذ‬
‫ذأرظؽا تٍُ كم يٍ انهعٍُ اإلَطانٍ وانكزَُىنٍ‪0‬‬
‫سُعهد أكثز قُى األؼًاض انذهُُح فٍ عُُاخ انغذاء انًهكٍ نطىائف انهعٍُ انكزَُىنٍ‬
‫انُؽم‬ ‫فٍ ؼٍُ ذأرظػ انًزكز انصاٍَ وانصانس تٍُ عُُاخ انهعٍُ اإلَطانٍ و سالنح‬
‫انًصزٌ انُقٍ فًُا عذا ؼايط ‪ -10‬هُذروكسٍ ‪ -2-‬دَسُُىَك انذٌ كاَد أعهً َسثح نه‬
‫فٍ عُُاخ انهعٍُ اإلَطانٍ شى سالنح انًصزٌ انُقٍ و أخُزاً انهعٍُ انكزَُىنٍ ‪ 0‬كذنك فقذ‬
‫كاَد أعهً َسثح نهؽايط انسُز‪ ،‬سُز‪ -9,12 -‬أوكرادَسُُىَك فٍ عُُاخ سالنح انُؽم‬
‫انًصزٌ انُقٍ ذثعها عُُاخ انهعٍُ انكزَُىنٍ شى انهعٍُ اإلَطانٍ‪.‬‬
‫انكهًاخ انذانح‪َ :‬ؽم انعسم‪ ،‬غىائف‪ ،‬غذاء يهكٍ‪ ،‬انُزقاخ انًطعىيح‪ ،‬هعٍُ‪ ،‬سالنح‪،‬‬
‫أؼًاض ايُُُح‪ ،‬أؼًاض دهُُح‬

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