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Sexing of bovine Shami embryos using PCR

تجنيس أجنة الأبقار الشامية باستخدام تقانة التفاعل التسلسلي البوليميرازي

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 Publication date 2013
  fields Animal Production
and research's language is العربية
 Created by Shamra Editor




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Thirteen viable and post - frozenthawed Shami cows embryos obtained by super-ovulation and artificial insemination technique in order to determine sex of embryos. Biopsies (1-2 cells) of embryos at morula or blastocyst stage were taken using micromanipulation under inverted microscope. It was concluded that aspiration of 1-2 blastomere (s) from embryos at morula or blastocyst stage was enough to determine sex of embryo using PCR and it is recommended that using this technique can be a useful tool in splitting and transferring of desired sex embryos in genetic improvement program.


Artificial intelligence review:
Research summary
تناولت الدراسة استخدام تقنية التفاعل التسلسلي البوليميرازي (PCR) لتحديد جنس أجنة الأبقار الشامية. تم جمع 13 جنينًا من أبقار شامية بعد تحفيزها بالإباضة المتعددة والتلقيح الصناعي. تم أخذ عينات من الأجنة في مراحل التويتة أو الكيسة الأريمية باستخدام إبرة رفيعة تحت المجهر المقلوب. تم استخدام 16 بادئًا لتضخيم الحمض النووي (DNA) المستخلص من الأجنة، وأظهرت النتائج أن أربعة من هذه البادئات (P1sex، P2sex، P1Aut، P2Aut) كانت فعالة في تحديد جنس الأجنة. أظهرت النتائج أن ظهور حزمة بحجم 141bp على هلامة الأغار يشير إلى أن الجنين ذكر، في حين أن غيابها يشير إلى أن الجنين أنثى. خلصت الدراسة إلى أن استخدام تقنية PCR يمكن أن يكون أداة فعالة في تحديد جنس الأجنة قبل نقلها، مما يساعد في تحسين الوراثة وزيادة الفاعلية الاقتصادية لبرامج نقل الأجنة.
Critical review
دراسة نقدية: تعتبر هذه الدراسة خطوة مهمة في مجال تحسين الوراثة للأبقار الشامية باستخدام تقنية التفاعل التسلسلي البوليميرازي (PCR). ومع ذلك، هناك بعض النقاط التي يمكن تحسينها. أولاً، عدد الأجنة المستخدمة في الدراسة كان محدودًا (13 جنينًا فقط)، مما قد يؤثر على تعميم النتائج. ثانيًا، نسبة نجاح التقنية كانت 70% فقط، مما يشير إلى وجود بعض التحديات التقنية التي قد تحتاج إلى مزيد من البحث والتطوير. ثالثًا، لم يتم التطرق إلى الجوانب الاقتصادية بشكل كافٍ، مثل تكلفة التقنية مقارنة بالفوائد المحتملة. على الرغم من هذه النقاط، فإن الدراسة تقدم مساهمة قيمة في مجال تحسين الوراثة للأبقار الشامية وتفتح الباب لمزيد من الأبحاث في هذا المجال.
Questions related to the research
  1. ما هي التقنية المستخدمة في تحديد جنس أجنة الأبقار الشامية في هذه الدراسة؟

    تم استخدام تقنية التفاعل التسلسلي البوليميرازي (PCR) لتحديد جنس أجنة الأبقار الشامية.

  2. كم عدد الأجنة التي تم استخدامها في الدراسة؟

    تم استخدام 13 جنينًا في الدراسة.

  3. ما هي البادئات التي كانت فعالة في تحديد جنس الأجنة؟

    كانت البادئات P1sex، P2sex، P1Aut، P2Aut فعالة في تحديد جنس الأجنة.

  4. ما هو المؤشر الذي يدل على أن الجنين ذكر؟

    ظهور حزمة بحجم 141bp على هلامة الأغار يدل على أن الجنين ذكر.


References used
Avery, B., A. Bak and M. Schmidt. 1989. Different cleavage rates and sex determination in bovine embryos. Theriogeno., 32: 139-147
Booman, P., L. Kruijt, R. Veerhuis, A. M. Hengst, M. Tiemann and F.E. Ruch.1989. Sexing bovine embryos with monoclonal antibodies against the H-Y antigen. Livestock Production Science, 23: 1–16
Cenariu, M., I. Groza, P. Emoke, L. Bogdan, I. Morar, S. Ciupe and R. Pop. 2011. Sexing of bovine embryos using polymerase chain reaction (PCR) and fluorescent In Situ hybridization (FISH). Romanian Biotechnological Letters, 16(2): 6055-6061
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Twelve growing Shami heifers available in Dier-Al-Hajar Station for improving Shami Cattle during 2005-2007 weighing 250±11 kg and 400±21 days old were used to monitore the growth of dynamic of ovarian follicles (daily numbers, size, length and nu mber of waves, rate of growth and regression) and characteristics of estrus cycle (length, time of dominancy, diameter of Graffian follicle and estrus and ovulatory times), length of corpus luteum (CL) during the estrous cycle. Heifers were maintained under uniform conditions and fed according to their daily growth. Transrectal Ultrasound Echo System (7.5 MHz probe) was used to study the ovarian structures and to monitor the daily development and regression of ovarian follicles, the formation and regression of corpus luteum in the left and right ovaries during the estrus cycle. Results indicated that the growth turnover of ovarian follicles was observed and developed in a wave pattern in all Shami heifers and no significant differences in the number and sizes of ovarian follicles were found between right and left ovaries. A cohort of 5 to 7 small follicles with a diameter 2-5 mm developed together and one of them was selected to continue growing and becomes dominant and later on being regressed or ovulated during the estrus cycle. One to five follicular waves were observed per cycle and each wave was characterized by the development of one large (dominant) follicle and a variable number of smaller follicles. The estrus cycle with one follicular wave occurred only in heifers exhibited the first post pubertal cycles with an average length equaled to 8.67±0.33 days. Frequency of estrus cycle according to the number of follicular waves was 8.4, 25.3, 44.2, 20 and 2.1% in cycles having 1, 2, 3, 4 and 5 follicular waves, respectively. Furthermore , the length of estrus cycle was also related to the number of follicular waves and it was 21.87±0.62, 22.44±0.74, 26.4±0.72, 26.50±3.50 days in cycles having 2, 3, 4 and 5 follicular waves, respectively with significant differences (P < 0.05) in cycles with 2 or 3 vs. 4 and/or 5 waves. The functional duration of CL was extended to 9, 12 and 15 days in a short (17-19 days), intermediate (20-24 days) and long estrus cycle (≥ 25 days). The maximum diameter of dominant follicles in ovulatory waves was significantly larger (P < 0.05) than that of dominant follicles in nonovulatory waves. In conclusion, this study showed that the development of ovarian follicles in Shami heifers occurs in waves and the most common pattern is three waves per estrous cycle.
This research was conducted I during 2009-2010 at Dier Alhajar station, General Commission for Scientific Agricultural Research to determine the length of calving interval (CI) and the effect of sire, calving year, calving season, parity and age o f cow and their interactions in Shami Cattle. It was concluded that CI can be reduced by better management and providing good and suitable feeding conditions.
This study was conducted at the biotechnology laboratory of AnimalWealth Research Administration, General Commission for Agricultural Research in order to compare the effect of different media on In vitro maturation (oocytes matured at 39 C° under 5% CO2 for 24 h), fertilization، and culture (7 days postfertilization) in humidified atmosphere 95% on bovine embryo production.
This study conducted at Deir Al-Hajar Station during the season 2010-2011 to describe the lactation curve of Shami cattle and determine factors affecting its shape compenents. 1120 records of 356 Shami cows collected between 1997 – 2010 were used. Data were subjected to general linear model, and the analysis of variance used to determine the effect of factors. Duncan test was used to compare the means using SAS system (1996) and Incomplete gamma functions were used to estimate the parameters of lactation curve (a, b and on the basis of daily yield of milk. Results indicated that the average values of the gamma parameters were 2.14 ± 0.01 kg for a (beginning of milking ), 0.61 ± 0.02 kg for b (increasing milk production up to peak) and - 0.23± 0.01 kg for c (decreasing milk production from peak to dry). The effect of calving year was significant (p<0.01) on a, and (p<0.05) on b and (p<0.001) on c. The effect of age at calving and sex of calf was significant (p<0.01) on a but non significant on b and c. Season of calving, parity, and interaction between age and parity was not significant on all parameters. It was concluded that improving the productive performance of Shami cattle, requires applying a long term genetic improvement program to select the best herd.
Experiment was conducted on 160 fertilized egg sterile, distributed in four groups of 40 eggs in each of them, eggs incubation in the hatchery for 12 h, and then go out and inject eggs experimental Groups (1-2-3) with a solution of tetracycline under the embryo disk.

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