The Effect of Sexing Process by Using Density Gradient Centrifugation Percoll and Frozen Method to Sperm Motility and Membrane Damage of Ongole Crossbred Bull

Abstract

Artificial insemination using semen Sexing aim to improve the genetic quality and to get calf with gender according to expectations. Sexing using density gradient centrifugation percoll and sedimentation egg whites have been successfully frozen. This study aims to determine the effect of sexing process using density gradient centrifugation percoll and Egg white sedimentation on motility and membrane damage spermatozoa. The material used is bull Filial Ongole semen have motility which has more than 65%. Treatment ; T0 = without sexing; T1 = Sexing. Each treatment was conducted repeat 10 times. The design used the Block Design and descriptive analytics. Media diluent used is andromed @ . Percentage motility by using a Light microscope magnification 100X. while damage to the membrane using Electrone Scanning microscope (SEM) with a magnification of 10,000 X. The result is the percentage motility ; T0 = 64,25 ± 3,94%; T1 After sexing , X-sperm and Y-sperm = 53± 7,93 % dan 48,55 ± 8,28%, . Motility before freezing at T0 = 59,75 ± 4,48%. T1 before Freezing , X-sperm and Y-sperm = 47 ± 6,13% dan 42,1 ± 7,36% and Post thawing motility X-sperm and Y-sperm T0 = 44,2± 2,79%, T1 = 31,45± 7,20% and 27,45± 8,69%. On Raw semen looked membrane intact spermatozoa. Y sperm after sexing were damaged membranes of the head and in spite of the sperm head to tail, whereas the X sperm membrane damage not appear. After freezing both the control and the results of many spermatozoa sexing damaged membranes and head with tail no intact.

Analysis of Spermatozoa Quality Using Percoll Density Gradient Centrifugation Through the Administration of Phospholipid + Egta

“…The criteria for good-quality fresh semen were as follows: per-ejaculate volume ranging from 5 -8 mL and concentrations between 700 and 2,000 million sperm per mL (Garner and Hafez, 2000;Berden and Fuquay, 1984). Also, semen with a motility percentage above 70% is more viable than that with lower motility (Susilawati et al, 2017). This has been supported by other studies: motility of at least 70% (Zenichiro et al, 2002), motility > 70% and pH level of 5.9 -7.3 (Hafez, 2000); viability > 80% (Ax et al, 2000) and membrane integrity level of 72.90% (Check et al, 1992).…”

“…This approach is easy in its implementation and application and a valid and inexpensive method. However, results from several recent studies indicate that the PDGC method may cause damage to spermatozoa membranes, which can further decrease the quality of spermatozoa (Susilawati et al, 2017).…”

Karakter Motilitas Spermatozoa Hasil Sexing pada Sapi Peranakan Ongole dengan Volume Awal yang Berbeda

“…Diungkapkan oleh Susilawati et al (2017) perlakuan sexing spermatozoa mengakibatkan penurunan kualitas spermatozoa. Hal inilah yang menjadikan dasar inseminasi buatan (IB) menggunakan semen sexing dobel dosis atau dua straw untuk satu kali IB.…”

Kualitas Semen Beku Sexing Sapi Peranakan Ongole Menggunakan Volume Semen Awal Yang Berbeda

“…Hasil penelitian Fernanda et al (2014) menunjukkan bahwa setelah thawing semen sexing, persentase motilitas spermatozoa X sebesar 35% dan spermatozoa Y sebesar 40%. Hasil penelitian Susilawati et al (2017) menyatakan bahwa motilitas spermatozoa X dan Y semen sexing SGDP post thawing masing-masing adalah 31,45±7,20% dan 27,45±8,69%. Hasil penelitian Mahfud et al (2019) menyatakan bahwa konsentrasi spermatozoa sexing beku sebanyak 12,125 juta/straw.…”