الفهرس 
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Abstract
Four experiments were conducted in this study during the period from October 2011 to August 2013 at the laboratory of physiology and biotechnology, Animal Production Department,Faculty of Agriculture, Minoufiya University, Egypt.1- First experiment:This experiment aimed to study the effect of season(breeding season and non-breeding season), reproductive status(Ovaries with CL and without CL) and site of the ovary (right and left) on: 1) some ovarian measurements (weight, volume, length and width). 2) morphological investigation of ovarian follicles and oocytes. 3) oocyte yield and recovery rate.1- Ovarian measurements:One hundred and ninety nine ovaries were collected from 102 Dromedary camels at random from El-Bassatein and El-Warraq slaughterhouses of Cairo, Giza Province, left and right ovaries were collected separately per donor and placed in punctured plastic bag immediately after slaughtering then placed into thermos in saline solution (0.9% NaCl) supplemented with antibiotics (100 IU penicillin and 100 μg streptomycin/ml) at 28-30°С. The ovarian measurements included the following, Ovarian weight and volume, Ovarian length and width. At the same time,the ovaries were examined for the presence of CL.2- Ovarian follicles and oocytes:The respective follicles were classified at counting into three categories depending on their surface diameter and the number of each category was recorded. The three categories were as follows: Small follicles (1 > 2 mm), Medium follicles (2-8 mm),Large follicles (> 8 mm).3- Oocyte yield and oocyte recovery rate:Oocytes were aspirated only from visible follicles having 2-8 mm in diameter (medium - sized follicles) using a 20-gauge hypodermic needle attached with a sterile disposable 10 ml syringe containing 2 ml harvesting medium. After aspiration,contents of each syringe were slowly dispelled into Petri dish for searching oocytes under a stereomicroscope. After oocytes collection, oocytes were counted and evaluated under a stereomicroscope in respect to both investment and ooplasm granulation.The oocytes were classified into four categories based on their cumulus investment as follows: Complete, Partial, Expanded,Denuded, fragment oocyte (parthenogenesis) and degenerated. At the same time, oocytes were classified into three categories based on their cumulus evenly granulated dark ooplasm as follows:Even, Uneven and Shrunken.The obtained results could be summarized as follows:1- Overall average of ovarian weight and size 5.6 gm and 5.1 cm3.It was affected significantly (P<0.01) by season, reproductive status and ovarian site.2- Ovarian size was associated with ovaries with CL (6.3 cm3) in left site (6.3 cm3) than those obtained during breeding season (4.9 cm3) without CL (4.6 cm3) in right site (4.2 cm3) with significant (P<0.01) differences.3- Length and width obtained of ovary with CL (3.9 and 2.8cm)or from the left ovaries (3.9 and 2.9 cm) was significantly higher than that obtained of ovary without CL (3.5 and 2.6 cm.,respectively) or from the right ovaries (3.3 and 2.5 cm,respectively). , oslAoverall average of ovarian length and width during breeding season (3.6 and 2.8 cm) and non-breeding season (3.6 and 2.7 cm), respectively.4- Small follicles represented the largest number and percentage (2451 and 70.1%), respectively of that count, however, medium follicles represented (979 and 28.0%) and the large follicle recorded the lowest number and percentage (66 and 1.99 %,respectively).5- The average number of MF and LF/ovary was significantly higher on of ovary without CL (5.2 and 0.4, respectively) than that recorded for ovary with CL (4.5 and 0.1), respectively camels. The average number of TVF and SF/ovary was insignificantly higher on ovary without CL (18.6 and 12.7,respectively) than that recorded for ovary with CL (16.1 and 11.5), respectively.6- Overall average of the oocyte yield/ovary was 8.8 with 50.3%
as overall average of recovery rate. The average of oocyte yield
per ovary and the percentage of recovery rate during breeding
season (10.7 and 63.7), respectively. was significantly higher
(P<0.01) than that obtained during non-breeding season (7.1
and 39.18 %), respectively.
7- The average of oocyte yield per ovary of ovary with CL (8.1)
was insignificantly lower than that oocytes yield/ovary from
Summary
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ovarian follicles of ovary without CL (9.3), and the percentage
of recovery rate of those animals was 50.6 and 50.1,
respectively without significant difference.
8- The average number of oocyte yield per ovary of left ovaries
was significantly higher (P<0.05) than that recorded for right
ones (10.17 vs. 7.50, respectively).
9- Overall average of oocyte investments is affected significantly
by the season and insignificantly by or the reproductive status
or site of the ovary.
10- None of the factors of season, reproductive status or site of the
ovary significantly affected the proportional distribution of
various grades oocyte ooplasm granulation.
2- Second Experiment:
This experiment was conducted to investigate the influence
of supplementation maturation medium with different types and
concentration 10, 20 % of camel serum (namely, Fetal dromedary camel
serum (FDCS), pre-mating dromedary she-camel serum (PMDSCS),
mating dromedary she-camel serum (MDSCS) and post-mating
dromedary she-camel serum (POMDSCS) on maturation rate throw
evaluation the cumulus cells expansion and stage of nuclear maturation of
camel oocytes. On the other hand, effect of season (breeding season and
non-breeding season) on maturation rate throw evaluation the cumulus
cells expansion and stage of nuclear maturation of camel oocytes.
The expansion degree of the cumulus-oocyte complex
(COCs) was classified as follow: Fully expanded (all cumulus cells were loosened), Partially expanded (the outer layer of cells
was loosened) and Not expanded.
It is worthy to note that, results of the oocytes nuclear
maturation stages were classified into two categories as follow:
Mature oocytes: included the oocytes nuclear maturation stages
of Telophase I and Metaphase-II (MII). And Immature oocytes:
included stages of nuclear maturation of Germinal vesicle (GV),
Germinal vesicle breakdown (GVBD), Metaphase-І (MІ),
Anaphase-I (AI) and Degenerated oocytes.
The obtained results could be summarized as follows:
1- supplementation the maturation medium with postmating camel
serum resulted in improving the average of matured camel
oocytes (57.0%) significantly (P<0.05) than that occurred when
the maturation.
2- Culturing camel oocytes in TCM media containing 20%
postmating camel serum yielded an insignificant increase in
the percentage of matured oocytes (59.3%) comparing with
that obtained with TCM medium containing 10% postmating
serum (53.7%) and a significant (P<0.05) improvement when
maturation TCM media contained 10% or 20% mating serum
(34.3 and 28.9%, respectively).
3- No significant effect of serum type on expansion rate. The
average of expansion rate of oocyte cultured in TCM media
containing premating, mating, postmating and fetal dromedary
camel serum was 84.4, 88.1, 84.3 and 84.4%, respectively, and
the average of expansion rate of oocyte cultured in TCM
media without serum supplementation was 84.6%. No significant difference between the percentage of matured
oocytes cultured in breeding or non-breeding season, with an
insignificant increase for the cultured oocytes of
breeding season ( 44.7%) than that noticed for the cultured
oocytes of non-breeding season (43.31 %).
5- No significant differences between the expansion rate of
cumulus cells of the oocytes cultured in breeding or nonbreeding
season with insignificant increase for the cultured
oocytes of breeding season than that reported for the cultured
oocytes of non-breeding season ( 86.94 and 84.81 %,
respectively).
Third Experiment:
This experiment was conducted to investigate:
1. The influence of supplementation of α-tocopherol (VE) at levels
of 0, 1 and 2 mM/ml, two concentrations of heparine (10 and 20
μg/ml) & two concentrations of cafaine (10 and 20 μg/ml) to
semen extender on the acrosme recation of epididmal camel
spermatozoa.
2. Effect of adittion of V.E at levels of 0, 1 and 2 mM/ml to the
extended epididymial camel semen with tris-egg yolk extender
and period of storage semen at 5oC for 96 hrs. on the motility
and liveability percentage of epididymial camel spermatozoa
Total of 32 testes of mature camel bulls were collected from
Kom-Hamada slaughterhouse, immediately, after animal
slaughtering and placed into plastic bag into icebox at 5°C, then
transport.In the laboratory, each testis was dissected away from its
tunica vaginalis and other extraneous tissue, then washed 3 times
by tap water and finally washed with ethyl alcohol (70%). Various
incisions in the tail of epididymis were performed with a scalpel
and then, by pressing that region manually the epididymal
spermatozoa samples were released and collected by aspiration
with sterile disposable (5 ml) syringe containing 2 ml semen
extender. The suspended or recovered semen samples were placed
in a 5 ml tube.
1- Sperm mtility and Sperm livabiity (%) examined with V.E at
levels of 0, 1 and 2 mM/ml to the extended epididymial camel
semen with tris-egg yolk extender and period of storage semen
at 5oC for 96 hrs.
2- Acrosome reaction examined with α-tocopherol (VE) at levels
of 0, 1 and 2 mM/ml, two concentrations of heparine (10 and
20 μg/ml) & two concentrations of cafaine (10 and 20 μg/ml)
to semen extender. The percentage of acrosome reacted
spermatozoa was calcualated for 100 spermatozoa selected at
random on each slide.The progression of the acrosome was
classified by estimating the degree of change in acrosome into
4 groups: 1- Ld: Live reacted 2- Dd: Dead reacted 3- La: Live
unreacted 4- Da: Dead unreacted sperm.
The obtained results could be summarized as follows:
1- The highest percentage of the reacted live sperm (40.3%)
occurred with supplementation the extended epididymial camel
semen with treatment of (H10 μg/ml + C10 μg/ml) in the presence of V.E at level of 1mM; however the lowest value of
reacted live sperm (29.3%) was registered with adittion of C
20μg/ml in the absence of the V.E.
2- Extended semen with H or C or H+C combination in the
presence of V.E resulted in improvement the acrosme reaction
process for the epididymial camel spermatozoa than that
occurred in the absecnce of V.E.
3- No significant effect observed either for V.E supplementing to
the extended semen or for incubation the extended semen at 5ºC
for 96 hrs. on the motility and liveability percentage of camel
epididymall spermatozoa
3- Fourth Experiment:
- The best results of the in vitro maturation rate of the camel
oocytes and in-vitro sperm capacitation experiments, this
experiment aimed to study the effect of supplementation the
embryo culture medium with 20% Post-mating serum on the
fertilization and cleavage rate.
- Embryo culture medium consisted of TCM-199 supplemented
with 100 IU/ml penicillin, 100 μg/ml Streptomycin and 20%
Post-mating. Cleavage and development to blastocyst stage was
recorded on days 4-7 of culture.
The obtained results could be summarized as follows:
higher total cleavage rate (39%) recorded for the cultured
oocyte supplemented with 20% Post –mating serum as compared
to that obtained from cultured oocyte without these supplements.And the percentage of oocytes reached to the morula stage was
35.4% and to the blastocyst stage was 3.7%. These results
indicated satisfied success rate of fertilization of in vitro matured
camel oocyte using epididymal spermatozoa of dromedary camel.