Separation of Bull Spermatozoa Bearing X- and Y-Chromosome by using Albumin Gradient and Swim-up Technique in Pesisir Cattle

Afriani, Tinda; Udin, Zaituni; Hellyward, James; Purwati, Endang; Rastosari, Adisti; Wahyudi, Dwiki

doi:10.17582/journal.jahp/2022/10.3.337.343

Cited by 3 publications

(4 citation statements)

References 13 publications

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“…These were smaller than the motility rates of 75% for X and 67% for Y observed by Agasi et al (2020) in Sumba Ongole cattle using albumin column. They were also slightly different from the motility rates of 58.33±10.32% for X and 60.83±13.57% for Y reported by Afriani et al (2022). This current study indicated a 20-30% decrease in the motility percentage of sexed and non-sexed spermatozoa.…”

Section: Evaluation Of Semen Quality After Sexing In Pesisir Bullscontrasting

confidence: 89%

“…The average quality of IPM in sexed Pesisir cattle bull spermatozoa was determined to be 55.58±1.01 for X and 59.35±7.78 for Y, as presented in Table 2. These results differed slightly from the 53.63±16.54% X and 52.76±8.86% Y reported by Afriani et al (2022). Furthermore, they were smaller than IPM percentages of 83.49% in the upper layer (X) and 83.62% in the lower layer (Y) obtained by Purwoistri et al (2013) from the Limousine bull spermatozoa sexed using the albumin gradient method.…”

Section: Evaluation Of Semen Quality After Sexing In Pesisir Bullscontrasting

confidence: 73%

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Molecular Verification of Spermatozoa Sexing Method in Pesisir Cattle Bull Using Bovine Serum Albumin Column

2023

Int J Vet Sci

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An accurate sexing process is crucial for preserving the purity of Pesisir cattle bulls and enhancing their productivity to meet the increasing beef demand in Indonesia. Therefore, this study aims to verify the separation of spermatozoa carrying X and Y chromosomes molecularly, using the Bovine Serum Albumin (BSA) column. The spin-column method was employed to isolate the separated X and Y spermatozoa. Amplification through PCR was performed using two sets of primers targeting the Sex-determining Region Y (SRY) gene, located on the Y chromosome, and the Ausal Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene on the X chromosome. The assessment of spermatozoa quality in coastal cattle bulls showed an average fresh semen volume, and sperm concentration to be 3±0.7mL, and 1.58±1.05x10 6 /mL, respectively. While the other parameters of assessment of spermatozoa quality including average motility, spermatozoa survival, sperm abnormalities, and mean progressive motility (MPU) were to be 80±10, 85.83±7.59, 6.36±3.50, and 79.81±5.97%, respectively. The quality evaluation after sexing included the average motility of spermatozoa with X at 56.66±11.54 and Y at 66.66±11.54%, as well as their average intact plasma membrane (IPM) being 55.58±1.01 and 59.35±7.78%, respectively. The quality of spermatozoa decreased by 20-30% after sexing. The results of spermatozoa separation with the 5% BSA column confirmed one GAPDH band (415bp), indicating the content to be X. In contrast, the 10% BSA column and the non-sexed spermatozoa exhibited two bands, SRY (318bp) and GAPDH (415bp), indicating a higher proportion of Y in the 10% BSA column. These demonstrated molecular verification of sexed spermatozoa using the 5 and 10% BSA column, enabling the separation of those carrying X and Y chromosomes in Pesisir bulls through the duplex PCR method.

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Section: Evaluation Of Semen Quality After Sexing In Pesisir Bullscontrasting

confidence: 89%

Section: Evaluation Of Semen Quality After Sexing In Pesisir Bullscontrasting

confidence: 73%

Molecular Verification of Spermatozoa Sexing Method in Pesisir Cattle Bull Using Bovine Serum Albumin Column

2023

Int J Vet Sci

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“…The application of spermatozoa sexing technology in local cattle breeds like Pesisir bulls (Bos indicus) has not been previously explored. This technology can be employed in artificial insemination to produce more male offspring, thereby enhancing the efficiency of artificial insemination programs (Afriani et al 2022). Yurnalis et al (2017) stated that the Pesisir bulls breed is an indigenous bull's breed in Indonesia known for its adaptation to challenging conditions, particularly in hot and humid climates and with limited access to nutritious feed, contributing to its ability to produce meat.…”

Section: Cite Thismentioning

confidence: 99%

Testing Motility Parameters of Post-Thawing Pesisir Bulls (Bos indicus) Semen with and without Sexing

2023

Int J Vet Sci

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This study aimed to test the motility parameters of frozen semen from Pesisir bulls with and without sexing. The research material consisted of semen from two Pesisir bulls. The data obtained were statistically analyzed using ANOVA. The results of this study showed that the motility percentage of Pesisir bulls ranged from 53.66 to 58.87%, and progressive motility ranged from 41.20 to 48.87%. The values of distance curve line (DCL) ranged from 24.05 to 28.24μm, distance average path (DAP) ranged from 24.05 to 24.35μm, and distance straight-line (DSL) ranged from 18.49 to 22.78μm. The values of velocity curve line (VCL) ranged from 93.95 to 112.97μm/s, velocity average path (VAP) ranged from 53.90 to 62.57μm/s, and velocity straight-line (VSL) ranged from 41.97 to 50.51μm/s. The values of linearity (LIN) ranged from 0.43 to 0.45, straightness (STR) ranged from 0.75 to 0.80, and wobble (WOB) ranged from 0.54 to 0.59. Meanwhile, the amplitude of lateral head (ALH) value was between 4.22 and 5.14μm, and the beat cross frequency (BCF) value ranged from 21.34 to 26.99Hz. The motility parameters, progressive motility, DAP, DSL, VAP, VSL, and LIN of post-thawing Pesisir bulls semen did not show significant differences (P>0.05) both with and without sexing. However, there were significant differences (P<0.05) in the parameters of progressive motility, DCL, VCL, STR, WOB, ALH, and BCF. However, for future research, it is expected that studies could involve more significant and more diverse sample sizes, as well as variations in cryopreservation techniques that may be employed. Additionally, further research could consider factors such as the age and health condition of Pesisir bulls, which could influence semen quality after thawing.

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“…Gunawan et al (2015) reported that spermatozoa separation with 5% and 10% BSA column, yielded 85% of resemblance to the sex gender from artificial insemination result. According to Afriani et al (2022) reported that the semen quality of Pesisir Bull after sexing by albumin gradient method were 58.33% motility on xchromosome bearing spermatozoa and 60.83% motility on y-chromosome bearing spermatozoa. Sexing will decrease the quality of the spermatozoa when the cells got separated due to physical and chemical effect.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Motility Characteristic of Pesisir Bulls Sexed Semen with Different Pre-Freezing Method Based on Computer Assisted Sperm Analyzer (CASA)

Wahyudi,

Udin,

Afriani

2023

BuletinPeternak

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This study aimed to find the best combination between pre-freezing method and layer of post sexing on semen quality of Pesisir Bulls. This study used 2 Pesisir Bulls. The semen was evaluated using Computer Assisted Sperm Analyzer (CASA). This study used factorial randomized block design with 2 factors. Factor A was sexed semen layer of BSA column with 2 level, whereas Factor B was pre-freezing method with 3 level. Analysis of Variance (ANOVA) were used in this study for data Analysis. According to the analysis of variance, there is no interaction of the combination of two factors on each paramters. Pre-freezing method showed highly statistically significant effect (P<0.01) on the quality of motility on upper layer and bottom layer which was about (52.86-72.93%; 52.32-65.50%) and about (40.17-56.99%; 40.33-53.18%) on progressive motility. The value of Distance Curve Line on (39.15-43.24 μm; 49.25-53.86 μm), Distance Average Path (22.38-24.60 μm; 26.52-29.94 μm), Distance Straight Line (17.32-19.07 μm; 20.93-24.25 μm). The value of Velocity Curve Line (88.67- 97.50 μm/s; 109.85-117.90 μm/s), Velocity Average Path (50.98- 55.63 μm/s; 59.32- 64.69μm/s), Velocity Straight Line (34.45-42.37 μm/s; 46.80-50.62 μm/s), Beat Cross Frequancy (23.91-25.17 Hz; 25.63-27.35 Hz), Straightness (0.75-0.77; 0.77-0.80), and Wobble (0.56-0.57; 0.53-0.55), highly statistically significant (P<0.01) on post sexing layers. Meanwhile, the value of Amplitude of Lateral Head (4.24-4.94 μm; 4.25-4.44 μm) showed highly significant effect (P<0.01) on factor A and factor B treatment, and value Linearity was about (0.42-0.44; 0.43-0.44) showed not significant effect (P>0,05) on every factor. According to the study, it can be concluded that the best treatment was treatment modified procedure layer by space the straw 16 cm every layers and The quality of post sexing X- and Y- spermatozoa motility effect on motility characteristic such as DCL, DAP, DSL, VCL, VAP, VSL, STR, WOB, ALH and BCF.

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Separation of Bull Spermatozoa Bearing X- and Y-Chromosome by using Albumin Gradient and Swim-up Technique in Pesisir Cattle

Cited by 3 publications

References 13 publications

Molecular Verification of Spermatozoa Sexing Method in Pesisir Cattle Bull Using Bovine Serum Albumin Column

Molecular Verification of Spermatozoa Sexing Method in Pesisir Cattle Bull Using Bovine Serum Albumin Column

Testing Motility Parameters of Post-Thawing Pesisir Bulls (Bos indicus) Semen with and without Sexing

Analysis of Motility Characteristic of Pesisir Bulls Sexed Semen with Different Pre-Freezing Method Based on Computer Assisted Sperm Analyzer (CASA)

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