Artificial insemination using sexing semen is expected to produce calves with the desired sex. One sexing sperm method is the percoll density gradient centrifugation method. This study aimed to determine the changes and the relationship between motility and acrosome reaction after sexing process using percoll density gradient centrifugation. The material used was semen of ±5 years old Ongole crossbred bull with a bodyweight of ±700 kg as many as three bulls with mass motility 2+ and individual motility 70%. The method used was to compare fresh semen with sexed semen after the cooling process. Parameters measured were motility characters using CASA analysis, which included motility parameters, progressive motility, capacitation, and no acrosome reaction. Statistical analysis used paired T-test to distinguish among fresh semen, after sexing and cooling process. In comparison, regression and correlation were used to analyze the relationship of capacitation and hyperactivation sperm with no acrosomal reaction with motility and progressive motility. The results showed that motility and progressive motility decreased after the sexing and cooling process. Meanwhile, the acrosomal reaction, capacitation, and hyperactivity increased.

ABSTRAK. Sexing spermatozoa bertujuan untuk mengatur jenis kelamin sesuai harapan. Salah satu metode sexing adalah dengan menggunakan Sentrifugasi Gradien Densitas Percoll. Tujuan dari penelitian ini adalah untuk mengetahui berbagai parameter motilitas menggunakan Computer-assisted Sperm Analysis dan proporsi spermatozoa X dan Y menggunakan metode Sentrifugasi Gradien Densitas Percoll dengan volume awal yang berbeda. Penelitian dilakukan di Loka Penelitian Sapi Potong Grati, Pasuruan, pada bulan Januari sampai Maret 2020. Materi yang digunakan adalah Semen Sapi Peranakan Ongole berumur ±5 tahun dan bobot badan ±700 kg sebanyak 3 ekor, motilitas masa ≥ 2+ dan motilitas individu ≥ 70%. Metode yang digunakan adalah eksperimental menggunakan Rancangan Acak Kelompok (RAK) dengan 3 perlakuan volume saat sexing yaitu P1= 1 ml, P2= 1,5 ml dan P3= 2 ml dengan ulangan 11 kali. Ulangan juga berfungsi sebagai kelompok (block). Hasil penelitian menunjukkan bahwa setelah dilakukan sexing dan pendinginan hingga 5oC persentase motilitas adalah P1:71,02+10,08 %; P2: 79,63+8,65 % dan P3: 83,38+6,67 %, sedangkan motilitas progresif pada P1: 47,68+8,71%; P2: 59,61+7,26 % dan P3: 62,21+6,74%. Curvilinear Velocity (VCL) pada P1: 50,9±7,73 µm/s, P2: 55,2±5,03 µm/s dan P3: 53,2±5,97%. Straight-line Velocity (VSL) pada P1: 20,8±8,19 µm/s; P2: 21,6±6,02 µm/s; P3: 22,1±5,77 µm/s, sedangkan Average Path Velocity (VAP) pada P1: 30,2±7,82 µm/s; P2: 32,5±6,14 µm/s dan P3 : 31,5±6,18 µm/s. Linearity (LIN) pada P1: 40,2±12,26%;P2: 39,1±10,31%;P3: 39,8±7,25%. Straightness (STR) pada P1: 67,2±11,20%; P2: 65,7±10,06%;P3: 67,2±7,92% sedangkan Wobble (WOB) pada P1: 59±9,49%; P2: 58,8±8,63% dan P3: 59,1±7,74%. Kesimpulan dari penelitian adalah motilitas dan motilitas progresif spermatozoa pada sampel dengan volume awal 2 ml lebih baik dibandingkan dengan sampel dengan volume awal 1 ml dan 1,5 ml. (Characteristics of Sexing Spermatozoa Motility in Ongole Cattle with Different Initial Volume) ABSTRACT. Sperm sexing is a technique of sorting a specific type of sperm cell to fertilize the egg cell. One of the sexing methods that can be used for spermatozoa sexing is percoll gradient density centrifugation. This research aims to find out various motility parameters using Computer-assisted Sperm Analysis and the proportion of spermatozoa X and Y using density gradient centrifugation method percoll with different initial volume. The study was conducted at Grati Beef Cattle Research Station, Pasuruan, from January to March 2020. Materials of the study were Semen of 3 Filial Ongole Cattles, aged ± 5 years, with ± 700 kg body weight, mass motility ≥ 2+, and individual motility ≥ 70%. The method used was experimental using a Randomized Group Design with 3 treatment volume when sexing i.e. T0= 1 ml, T1= 1.5 ml dan T2= 2 ml with repetition 11 times. The repetition also functions as groups (blocks). The results showed that after sexing and cooling up to 5oC, the percentage of motility is T0:71.02+10.08 %; T1: 79.63+8,65 % and T2: 83.38+6.67 %. Progressive motility on T0: 47.68+8.71%; T1: 59.61+7.26 % and T2: 62.21+6.74%. Curvilinear Velocity (VCL) on T0: 50.9±7.73 µm/s, T1: 55.2±5.03 µm/s and T2: 53.2±5.97%. Straight-line Velocity (VSL) on T0: 20.8±8.19 µm/s; T1: 21.6±6.02 µm/s; T2: 22.1±5.77 µm/s. The Average Path Velocity (VAP) on T0: 30.2±7.82 µm/s; T1: 32.5±6.14 µm/s and T2 : 31.5±6.18 µm/s. Linearity (LIN) on T0: 40.2±12.26%; T1: 39.1±10.31%;T2: 39.8±7.25%. Straightness (STR) on T0: 67.2±11.20%; T1: 65.7±10.06%; T2: 67.2±7.92%. Wobble (WOB) on T0: 59±9.49%; T1: 58.8±8.63% and T2: 59.1±7.74%. The conclusion of the study is the motility and progressive motility of spermatozoa in samples with an initial volume of 2 ml is better than samples with an initial volume of 1 ml and 1.5 ml.