ANALISIS HASIL SEXING SPERMATOZOA SAPI SIMENTAL METODE KOLOM BOVINE SERUM ALBUMIN DARI DUA MACAM PENGENCER DENGAN TEKNIK FLOWCYTOMETRY

EKOWATI, INDAH AGUSTINA and Setiawan, Arum and Abrar, Arfan (2023) ANALISIS HASIL SEXING SPERMATOZOA SAPI SIMENTAL METODE KOLOM BOVINE SERUM ALBUMIN DARI DUA MACAM PENGENCER DENGAN TEKNIK FLOWCYTOMETRY. Masters thesis, Sriwijaya University.

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Abstract

Artificial Insemination (AI) is a reproductive technology to improve the genetic quality of livestock with the aim of helping farmers obtain superior seeds. The IB program is developed through the use of semen resulting from the process of separating X and Y chromosomal spermatozoa or sexing semen. IB with sexed frozen semen can increase efficiency in animal husbandry. Female cows inseminated with spermatozoa carrying the X chromosome will produce female calves and spermatozoa carrying the Y chromosome will produce male calves. In the process of separating X and Y chromosomes, there are various methods used, including the albumin column method, centrifugation with percol density gradient, electrical method, electroforesis, swim up, and sephadex filtration. To determine the level of purity of X and Y chromosomes of sexed spermatozoa quickly, the Flowcytometry technique has been developed using the Flowcytometer tool. Based on this, this study was conducted with the aim of confirming the spermatozoa sexing technique of Bovine Serum Albumin column method for dissemination purposes, evaluating the quality of sexed frozen semen and analyzing the success rate of sexing with Flowcytometry technique. This research was conducted at the Laboratory of the Center for Breeding and Forage Animal Feed (BP-HPT) Sembawa and continued at the Genomics Laboratory of the National Research and Innovation Center (BRIN) Bogor. The cattle used in this study were Simental cattle. The sexing method used was the albumin column method. The separation media used were Bovine Serum Albumin (BSA) 5% and 10%, while the diluent media used were Egg Yolk Tris (TKT) and Bovifree (BF). The research was carried out in stages, namely sexing spermatozoa using the Albumin column method, testing the quality of sexed spermatozoa and analyzing the purity of X and Y chromosomes of sexed spermatozoa using Flowcytometry technique. The results showed that the evaluation results of fresh semen spermatozoa of Simental cattle met the standards to proceed to the sexing process. After the sexing process, a second evaluation was carried out which included motility, viability and abnormality. Frozen semen without sexing has an average motility of 50.5% + 1.76, viability of 62.54% + 0.57 and abnormality of 4.8% + 0.41. While frozen semen sexing has an average motility of 41.25 + 6.85, viability of 57.97 + 0.57 and abnormality of 3.93 + 0.18. The results of analysis with Flowcytometer showed the average percentage of X and Y chromosomes in the upper column with TKT diluent X chromosome was 74.69% + 4.77 and Y chromosome was 23.14% + 4.10. While in the lower column X chromosomes were 31.50% + 5.74 and Y chromosome was 65.72% + 5.75. With BF diluent, the average percentage of X and Y chromosomes in the upper column is X chromosome of 70.99% + 13.16 and Y chromosome of 25.32% + 13.21. While in the lower column the X chromosome was 30, 21 + 8.45 and the Y chromosome was 66.73% + 8.96. Based on the results of the study, it can be seen that the use of diluent media affects the quality of sexing frozen semen due to the composition of the ingredients contained in the diluent media. The results of flowcytometer analysis, on sexing bovine serum albumin (BSA) column method with two kinds of diluent, showed the presence of unwanted chromosome contamination. Both for sexing upper column spermatozoa (X) with TKT and BF diluent media and lower column spermatozoa (Y) with TKT and BF diluent media. From the results of this study it is necessary to conduct further research to compare the quality of spermatozoa resulting from sexing with other diluents and methods and further research to prove the success of the results of X and Y chromosome separation with the implementation of insemination in the field.

Item Type:	Thesis (Masters)
Uncontrolled Keywords:	SEXING SPERMATOZOA
Subjects:	Q Science > QR Microbiology > QR171 Microorganisms in the animal body
Divisions:	08-Faculty of Mathematics and Natural Science > 46101-Biology (S2)
Depositing User:	Indah Agustina Ekowati
Date Deposited:	26 Jul 2023 02:40
Last Modified:	26 Jul 2023 02:40
URI:	http://repository.unsri.ac.id/id/eprint/121844

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