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Background and Aim: The production of male calf beef cattle is an agricultural innovation needed to increase the farm's productivity as a provider of meat sources. This study aimed to determine the sex ratio of the offspring of cows inseminated with Y-bearing sperm enriched by Percoll density gradient centrifugation and swim-up, combined with delayed fixed-time artificial insemination (FTAI). Materials and Methods: Ejaculates of Simmental bulls were divided into four equal portions and grouped as T0 (control, non-sexed semen), T1 and T2 were sexed semen using Percoll density gradient centrifugation three and five levels, respectively, and T3 was sexed semen using swim-up. After the sex was sorted, the semen was diluted in a tris-egg yolk extender, packaged in French mini-straws containing 50 million live sperm cells, and frozen. Pre-sexed, post-sexed, and post-thawed spermatozoa were evaluated based on progressive motility, viability, intact plasma membrane, and abnormality. The post-thawed semen of T0 was artificially inseminated to recipient cows at 12 h after onset of estrus (not delayed FTAI). Meanwhile, the delayed FTAI was conducted 18-20 h after onset of estrus using the T0, the best of T1 and T2, and the T3 post-thawed semen. Results: The Percoll density gradient centrifugation reduced motility, viability, and intact plasma membrane but increased sperm abnormalities. Meanwhile, the swim-up process increased motility, viability, and intact plasma membrane of sperm cells but decreased sperm abnormalities. Post-thawed semen decreased motility, viability, and intact plasma membrane of sperm cells but increased sperm abnormalities. The sex ratio of the Simmental crossbred offspring was 96.08% and 100% in T1 and T3, respectively, compared to 48.25% and 67.39% in T0 not delayed and delayed FTAI, respectively. Conclusion: The Percoll density gradient centrifugation and swim-up methods are prospective for obtaining male offspring.
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BACKGROUND AND AIM: The implementation of artificial insemination (AI) is one of the strategies to use superior male semen optimally to improve the genetic quality of livestock. One of the factors that influence AI is a fertility-associated antigen (FAA). This research aimed to examine the effects of FAA extracted from the accessory sex glands of a bull from a slaughterhouse that was added in bull semen freezing medium to increase cattle (bull) fertilization. MATERIALS AND METHODS: This research used a randomized complete block design. It consisted of two research phases, namely, explorative and experimental phases. The first phase involved determining the FAA molecular weight using the SDS-PAGE method, and the second phase consisted of laboratory and field testing, including testing the quality of frozen semen supplemented with FAA extracted from the accessory glands of a bull's genital organ from a slaughterhouse with various doses (0, 5, 10, and 15 µg in every 200 million progressively motile spermatozoa). RESULTS: The results showed that the percentages of bull sperm motility between the groups without and with the additional administration of FAA with a dose of 5 µg did not significantly differ. However, there was a difference between the groups without and with the additional administration of FAA with doses of 10 and 15 µg. After further testing, the highest percentage of sperm progressive motility occurred at a dose of 15 µg/200 million progressively motile spermatozoa (P3), which was equal to 2.59±46.88b (%). CONCLUSION: This research found that not all of the accessory glands (seminal vesicles) of bulls taken from the slaughterhouse contain the FAA. An FAA level between the accessory glands (seminal vesicles) of one cattle to another is different. The addition of the FAA protein from the accessory sex glands of a bull's organ in cattle semen can improve fertility by increasing the percentage of viability, motility, intact plasma membrane of spermatozoa, and pregnancy rate of bulls and decreasing the sperm capacitation post-thawing.
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AIM: The aim of this study was to know crude guava leaf tannins effect on motility, viability, and intact plasma membrane of stored spermatozoa of Etawa crossbred goats. MATERIALS AND METHODS: Macroscopic assessment of normal Etawa crossbred semen was followed by dilution with a glucose solution at a 1:10 ratio to increase volume. The diluted semen was treated by adding crude guava leaf tannins into 1 ml of the semen glucose diluent, and five treatments were obtained, namely, control group (C), with no added tannins; treatment Group 1 (T1), with 3%; treatment Group 2 (T2), with 6% tannins; treatment Group 3 (T3), with 12% tannins; and treatment Group 4 (T4), with 24% tannins. Each treatment used five replications. Then, microscopic analysis of the treated and control semen was carried out after 15 days of storage at 4-5°C temperature. The parameters observed were motility, pH, viability, abnormality, and intact spermatozoa plasma membrane. RESULTS: The spermatozoa motility in Group C was the highest (76.60±1.47). The motility in Group T1 did not differ from that in Group C, but was different and higher than that in Groups T2, T3, and T4. The pH of Group C tended to be acidic after 15 days of storage (4.78±0.01) as compared to the initial pH of fresh semen (6.76±0.12). The pH in Group C did not differ from that in the Groups T1 and T2, but differed from that in the T3 and T4 groups; the pH in the T3 and T4 groups was similar. The viability of spermatozoa in the T1 group was higher than that in all treatments (64.60±2.76); the lowest values were found in Group C (28.94±1.02). Group C had the lowest number of normal spermatozoa, with a mean of 72.58±3.48. The total number of abnormalities in the T2 group did not differ from those in the T3 group, and abnormalities in the T4 group did not differ from those in Group C, which exhibited the highest abnormalities in the head, neck, and tail. The most significant decrease was observed in the intact plasma membrane of spermatozoa on addition of 12% and 24% crude guava leaf tannin in glucose diluents. CONCLUSION: The addition of 3% crude guava leaf tannin to crossbred Etawa goat semen diluted with glucose diluent and stored for 15 days at 4-5°C resulted in a significant effect on spermatozoa motility, viability, and intact plasma membrane, whereas the administration of 24% crude guava leaf tannin resulted in low live percentage of spermatozoa.
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AIM: Biotechnological culture of hypoxia-conditioned (CH) rat mesenchymal stem cells (rMSC-CH) for testicular failure therapy with low libido improves the functional outcome of the testicle for producing spermatogenic cells and repairs Leydig cells in rats (Rattus norvegicus). MATERIALS AND METHODS: In the first group (T1), rats with testicular failure and low libido were injected with normoxia-conditioned (CN) rMSCs (21% oxygen); in the second group (T2), rats with testicular failure and low libido were injected with rMSC-CH (1% oxygen); in the negative control group (T-), rats with normal testis were injected with 0.1 mL phosphate-buffered saline (PBS); and in the sham group (TS), rats with testicular failure and low libido were injected with 0.1 mL of PBS. RESULTS: Vascular endothelial growth factor expression, as the homing signal, in the groups T2, T-, T1, and TS was 2.00±0.5%, 2.95±0.4%, 0.33±0.48%, and 0±0%, respectively. The number of cluster of differentiation (CD)34+ and CD45+ cells in the groups T- and TS was <20%, whereas that in T1 and T2 groups was >30% and >80%, respectively, showing the mobilization of hematopoietic stem cells (HSCs). The number of spermatogenic cells (spermatogonia, primary spermatocytes, secondary spermatocytes, and spermatid) decreased significantly (p<0.05) in TS compared with that in T-, T1, and T2, whereas that in T2 did not show a significant (p>0.05) decrease compared to that in T-. The improvement in libido, based on the number of Leydig cells producing the hormone testosterone for libido expression, did not increase in T1, whereas T2 was able to maintain the number of Leydig cells significantly compared to that between TS and T1. CONCLUSION: rMSC-CH culture for testicular failure with low libido showed improvement in the functional outcome of the testicle and in repairing Leydig cells.
