Academic literature on the topic 'Cattle Spermatozoa Storage'

The objective of this research was to examine the effectiveness of coconut water with various egg yolk concentrations in maintaining the viability of epididymal spermatozoa of crossbreed cattle preserved at 5oC. Five testis with epididymides of crossbreed cattle were obtained from slaughterhouse. Epididymal spermatozoa was collected by the combination of slicing, flushing and tissues pressure methods of cauda epididymides with saline solution (0.9% NaCl). Collected-spermatozoa was equally divided in volume into four tubes and diluted with lactose extender containing 20% egg yolk (control), 90% coconut water + 10% egg yolk (CWEY10), 85% coconut water + 15% egg yolk (CWEY15), and 80% coconut water + 20% egg yolk (CWEY20), repectively. Diluted-spermatozoa was stored in refrigerator at 5oC. Quality of dilutedspermatozoa including percentages of motile spermatozoa (MS), live spermatozoa (LS), spermatozoa with intact plasma membrane (IPM) were evaluated every day during four days of storage. Data were analyzed by using completely randomized design with four treatments and five replicates. Means were compared with significant difference test at 0.05 significant level. Results of this study showed that mean of spermatozoa concentration, percentage of MS, percentage of LS, percentage of spermatozoa abnormal, and percentage of IPM of crossbreed cattle fresh epididymal spermatozoa were 1,414 million cell/ml, 72%, 85%, 9%, and 90%, respectively. At day-4 of the storage, percentages of MS, LS, and IPM of control (43, 52.2, 59.2%) and CWEY20 (42, 52, 59%) were significantly (P<0.05) higher than CWEY10 (33, 45.4, 52.8%) and CWEY15 (37, 50, 54.6%). In conclusion, lactose and CWEY20 extenders could be maintaining the quality of epidydimal spermatozoa of crossbreed cattle for three days preservation at 5oC and two days for CWEY10 and CWEY15.

Artificial insemination with sexed semen, in vitro fertilisation and intracytoplasmic sperm injection have been used to reproduce animals, but often not as successfully as natural mating. Learning more about how spermatozoa normally interact with the female tract can provide inspiration for developing improvements in assisted reproduction. The present review focuses on Bos taurus, because more is known about this species than others. At coitus, bull spermatozoa are deposited into the anterior vagina, where they rapidly enter the cervix. Cervical mucus quickly filters out seminal plasma from spermatozoa, unlike most assisted reproduction protocols. Spermatozoa that reach the uterus may require certain cell surface proteins to swim through the uterotubal junction. Shortly after passing through the junction, most spermatozoa are trapped in a storage reservoir by binding to oviducal epithelium, in the case of cattle via bovine seminal plasma (BSP) proteins coating the sperm head. As ovulation approaches, spermatozoa capacitate and shed BSP proteins. This reduces sperm binding to the epithelium and releases them from storage. Motility hyperactivation assists spermatozoa in leaving the storage reservoir, swimming through oviducal mucus and the cumulus oophorus, and penetrating the oocyte zona pellucida. Chemotactically regulated switching between asymmetrical (i.e. hyperactivated) and symmetrical flagellar beating may also guide spermatozoa to the oocyte.

Background: Pasundan cattle are local cattle native to Indonesia. One way to conserve beef cattle genetics is to use Artificial Insemination technology. The success of Artificial Insemination can be influenced by the quality of semen. Purpose: To determine factors affecting fresh semen quality in Pasundan cattle at UPTD BPPIBTSP Ciamis. Methods: The data were obtained through observations on seven Pasundan bulls in March 2021 towards fresh semen quality and some factors influencing it. The Pasundan bulls observed were seven productive males. Results: The fresh semen quality of Pasundan cattle, such as volume, color, and pH, showed good result,s but the average consistency and concentration of spermatozoa were still below the standard. The factors that can affect the fresh semen quality are the breed of beef cattle, age, body weight, feed, season, exercise, and frequency of semen storage. Conclusion: The determining factors that can cause the consistency and concentration of Pasundan cattle’s spermatozoa at UPTD BPPIBTSP Ciamis are feed and season.

ABSTRAK digunakan sebagai salah satu bahan pengencer semen pengganti kuning telur ayam. Penelitian ini bertujuan untuk menguji pengaruh sari kedelai dalam pengencer Tris terhadap motilitas dan daya hidup spermatozoa sapi limousin yang dipreservasi pada suhu 5oC. Semen ditampung dengan vagina buatan. Semen segar yang memenuhisyarat dibagi kedalam empat buah tabung reaksi yang masing-masing berisi pengencer perlakuan, yakni: 80% pengencer dasar Tris + 20% kuning telur (Tris), 97% pengencer dasar Tris + 3% sari kedelai (SK3), 95% pengencer dasar Tris + 5% sari kedelai (SK5), dan 93% pengencer dasar Tris + 7% sari kedelai (SK7). Semen yang telah diencerkan disimpan di dalam refrigerator pada suhu 5oC, dan dievaluasi motilitas dan viabilitas spermatozoa setiap hari hingga hari kelima. Hasil penelitian menunjukkan bahwa motilitas dan viabilitas spermatozoa di dalam pengencer Tris nyata (P<0,05) lebih tinggi dibandingkan dengan spermatozoa di dalam pengencer SK3, SK5, dan SK7 selama empat hari penyimpanan. Berdasarkaan hasil penelitian dapat disimpulkan bahwa pengencer Tris-kuning telur lebih baik dalam mempertahankan motilitas dan daya hidup spermatozoa sapi limousin dibandingkan dengan pengencer Tris-sari kedelai yang dipreservasi pada suhu 5°C. Pengencer Tris dengan konsentrasi 3% sari kedelai lebih baik dibandingkan dengan konsentrasi 5% dan 7%.Kata Kunci : sapi limousine, sari kedelai, semen, trisABSTRACTSoybean contains anlecithin (phosphatidyl choline), that has the potential to be used as substitute for chicken egg yolk as one of the semen extender compound. The aim of this research was to examine the effect of soybean juice in Tris extender on the motility and viability of Limousin cattle spermatozoa preserved at 5oC. Semen was collected using artificial vagina. Fresh semen was divided into four tubes containing a treatment extender, i.e. 80% Tris base extender + 20 egg yolk (Tris), 97% Tris-based extender + 3% soybean juice (SJ3), 95% Tris-based extender + 5% soybean juice (SJ5), 93% Tris-based extender + 7% soybean juice (SJ7), respectively. Diluted-semen was preserved in refrigerator at 5oC, and evaluation of spermatozoa motility and viability were conducted on daily basis up to five days. The result showed that percentages of motility and viability of spermatozoa in Tris-yolk extender were significantly (P<0.05) higher than spermatozoa in SJ3, SJ5, and SJ7extenders during four days of storage. In conclusion, Tris-yolk extender is better than Tris-soybean juice in maintaining the spermatozoa motility and viability of Limousin cattle preserved at 5°C. Tris extender containing 3% soybean juice is better than 5% and 7%.Keywords: limousin cattle, semen, soybean juice, tris

Semen storage in cold temperatures might cause an increase in reactive oxygen species (ROS) production. This condition resulted in spermatozoa damage and quality decrease. This study was conducted to investigate the effects of α-tocopherol supplementation in diluents on the motility, viability, and plasma membrane integrity of Simmental bull spermatozoa after cooling. Semen samples were diluted in skim milk egg yolk supplemented with 0, 0.5, 1.0, and 1.5 mM α-tocopherol respectively for control, Tl, T2, and T3. Spermatozoa were evaluated for their motility, viability, and membrane integrity in cooling temperature (5°C). The daily evaluation showed that 1.5 mM α-tocopherol was the best in maintaining motility, viability, and plasma membrane integrity, while 1.0 mM α-tocopherol was only good for maintaining viability. Therefore, it can be concluded that α-tocopherol at the concentration of 1.5 mM was an efficient antioxidant supplement for Simmental cattle semen in skim milk egg yolk diluent.

The aim of this research was to know the effectiveness of guava filtrate supplementation in coconut water- egg yolk dilution on quality of liquid semen stored at 5oC of Bali cattle. Semen collected from a five year old Bali cattle using artificial vagina. Semen of good quality were kept in six tubes based on treatment then stored at 5oC. Treatments of the research were P0 : coconut water 80% + egg yolk 20% without guava filtrate; P1 : coconut water 80% + egg yolk 20% + 0.8% guava filtrate; P2 : coconut water 80% + egg yolk 20% + 0.9% guava filtrate; P3 : coconut water 80% + egg yolk 20% + 1.0 % guava filtrate; P4 : coconut water 80% + egg yolk 20% + 1.1 % guava filtrate and P5 : coconut water 80% + egg yolk 20% + 1.2 % guava filtrate. Each treatment was replicated 8 times making 48 experimental units. Results of the study showed that percentage mean of motility, viability, MPU, and TAU of spermatozoa after three days storage for P0 were : 42.20%, 41.85%, 39.08% and 40.90%; P1 : 50.40%, 53.89%, 52.99% and 54.67%; P2 : 54.67%, 56.97%, 54.51% and 54.36%; P3 : 17.00%, 29.96%, 29.64% and 29.64%; P4 : 23.38%, 24.64%, 21.06% and 24.45%Jurnal Veteriner Maret 2019 Vol. 20 No. 1 : 20 -29 pISSN: 1411-8327; eISSN: 2477-5665 DOI: 10.19087/jveteriner.2019.20.1.20 Terakreditasi Nasional, Dirjen Penguatan Riset dan Pengembangan, online pada http://ojs.unud.ac.id/index.php/jvet Kemenristek Dikti RI S.K. No. 36a/E/KPT/201621PENDAHULUAN Salah satu solusi yang dapat digunakan untuk pengembangan program Inseminasi buatan (IB) secara cepat dan mudah pada sapi bali adalah penggunaan semen cair. Penggunaan semen cair dapat meningkatkan kinerja IB pada sapi bali di Nusa Tenggara Timur (NTT). Keunggulan lain semen cair dapat diproduksi menggunakan bahan pengencer herbal berbasis bahan lokal dan peralatan yang sederhana serta mudah diperoleh dan tidak tergantung dengan persediaan nitrogen cair. Hasil akhir dari metabolisme spermatozoa adalah terbentuknya radikal bebas berupa derivat oksigen di antaranya adalah single1 oksigen (1O2), tripel1 oksigen (3O2), superokside anion (O2-), hidroksil radikal (OH) dan nitrit oxide (NO-) yang semuanya disebut radical oksigen species (ROS). Single1 oksigen dapat merusak ikatan rangkap pada asam lemak sehingga dapat menyebabkan kerusakan Deoxyribo Nuclead Acid (DNA) dan protein. Single1 oksigen bila bereaksi dengan asam amino histidin akan membentuk enzim yang dapat menyebabkan denaturasi protein. Kerusakan spermatozoa pada penyimpanan suhu 5%C akibat radikal bebas dan cold shock inilah merupakan penyebab utama disfungsi semen (Sharma et al., 2000). Oksidasi fosforilasi yang terganggu menyebabkan peningkatan radikal bebas dalam semen. Kadar radikal bebas yang terganggu menyebabkan peningkatan radikal bebas dalam semen. Kadar radikal bebas yang tinggi dalam sel dapat mengoksidasi lipid, protein dan DNA. Lipid membran plasma semen memiliki fosfolipid dengan kadar yang tinggi menyebabkan semen rentan terhadap radikal bebas (Sanoeka dan Kurpisz, 2004). Antioksidan bertindak mengikat asam lemak tak jenuh dan mencegah terjadinyareaksi berantai. Pada proses penyimpanan semen akan terjadi kerusakan membran plasma spermatozoa akibat terbentuknya perioksidasi lipid. Antioksidan-pemutus rantai seperti yang terkandung dalam jambu biji dapat menghambat perioksidasi lipid dalam membran melalui radical peroxyl (RO) dan alkoxyl (ROO) pengurai. Pengunaan jambu biji yang difilter dalam pengencer air kelapa kuning telur dapat menjaga kualitas spermatozoa (motilitas, keutuhan akrosom, viabilitas dan morfologi spermatozoa) semen cair sapi bali selama penyimpanan pada suhu 5%C. Dosis jambu biji yang difilter yang terbaik dalam pengencer air kelapa kuning telur, akan terbaik pula dalam mempertahankan kualitas spermatozoa sampai tujuan IB. Adapun tujuan penelitian ini adalah menguji berbagai level pemberian filtrat jambu biji (FJB) dalam pengencer air kelapa kuning telur terhadap motilitas, viabilitas, membran plasma utuh (MPU) dan tudung akrosom utuh (TAU) spermatozoa sapi bali yang disimpan pada suhu 5%C.METODE PENELITIAN Penelitian ini telah dilakukan di Laboratorium Reproduksi milik Yayasan Wiliams dan Laura yang berlokasi di Tilong, Desa Oelnasi, Kec. Kupang Tengah, Kab. Kupang, Nusa Tenggara Timur, dan berlangsung selama delapan bulan. Materi yang digunakan dalam penelitian ini adalah semen sapi bali yang ditampung dari satu ekor sapi bali jantan berumur lima tahun milik Yayasan Williams dan Laura yang telah dilatih, memiliki performans yang baik, dan organ reproduksi normal. Pakan yang diberikan adalah hijauan berupa rumput dan legum dan pemberian konsentrat secukupnya (dedak padi dan jagung giling).and P5 : 9%, 21.25%, 17.56% and 19.30%. Result of statistical analysis showed that there were a significant effect (P<0.05) between treatment on motility, viability, MPU and TAU of spermatozoa of Bali cattle till the third day of storage. It can be concluded that the supplementation of guava filtrate 0.9% in dilution of coconut water 80% - egg yolk 20% had been able to maintain motility, viability, MPU and TAU of spermatozoa of Bali cattle till the third day of storage at 5oC.

The endocannabinoid system (ECS) has been found in reproductive cells and tissues in several mammals. Spermatozoa are able to respond to anandamide, and the oviduct is able to synthesize and modulate the concentration of this endocannabinoid along the isthmic and ampullary regions. The main aim of this study was to understand whether the ECS has a role during sperm storage and release within the oviduct in cattle. Data showed that 1) the endocannabinoid receptors 1 and 2 (CB1 and CB2) are present in bovine spermatozoa both in the initial ejaculate and in spermatozoa bound to the oviduct in vitro; 2) CB1 receptor is still detectable in spermatozoa released from the oviduct through penicillamine but not in those released through heparin; 3) arachidonylethanolamide (AEA) does not affect sperm viability, whereas it depresses sperm progressive motility and kinetic values; 4) sperm–oviduct binding and release in vitro are not influenced by AEA; 5) AEA depresses sperm–zona pellucida (ZP) binding; 6) binding of heparin-capacitated spermatozoa to the ZP is not affected by AEA; 7) N-acylphosphatidylethanolamine-selective phospholipase D, the main enzyme involved in anandamide synthesis, is expressed in oviductal epithelial cells. In conclusion, secretion of AEA from epithelial cells might contribute to the oviduct sperm-reservoir function, prolonging the sperm fertile life through the depression of motility and capacitation. Capacitation signals, such as heparin, that promote sperm release, might remodel the sperm surface and cause a loss of the sperm sensitivity to AEA.

Despite being subject to prior assortment, frozen bull sperms commercialized for artificial insemination may present certain morphological defects. The present study aims (i) to assess the artificial insemination success of the most common cattle breeds in Algeria and (ii) to evaluate the possible effects of commercialized bull’s semen quality on this operation. Artificial insemination was assessed through four years of field monitoring by inseminating different cattle breeds of normal fertility. However, semen quality was evaluated using light microscopy by measuring viability, motility, and morphological abnormalities of spermatozoa. Field study revealed a high percentage of normal calving in red and white Holstein breed (44.83 %) against the high percentage of embryonic mortality (46.43 %) and calving with a malformation (10.71 %) in Montbéliarde breed. Semen quality assessment revealed that sperm viability and motility were higher in Holstein breeds than in Montbéliarde. Furthermore, significant differences between semen bulls were found in the proportion of abnormal spermatozoa; a higher rate of sperms with the abnormal head was observed in the black and white Holstein breed (69.3±10.98 %). However, the percentage of abnormal sperms with tail defects was higher in the Montbéliarde breed (67.5±10.74 %). The lousy quality of the selected semen and/or the poor handling and storage of frozen semen constitute a determinant factor that hinders the success of artificial insemination in the arid region of Tiaret (Algeria).

SummaryThe zona pellucida of mammalian oocytes stored in highly concentrated solutions of neutral salts is known to retain its biological and biochemical properties. However, the zona may become resistant to sperm penetration as the storage period is increased. In cattle and hamsters, the penetrability of zonae of salt-stored oocytes was restored or increased by treating the oocytes with moderate heat without altering the gross morphology of the zona. Although this technique did not work for salt-stored human ova, this may have been due to the use of so-called inseminated- unfertilised ova which (1) may have been fertilised but failed to activate, or (2) were not fertilised because of functionally defective zonae.

The competitiveness of buffalo breeding will depend on the utilization of reproductive biotechnologies that allows acceleration of genetic progress. A major factor hampering the efficiency of both artificial insemination and in vitro embryo production programs in this species is male hypofertility. Reports for several species suggest that seminal plasma contains factors that influence male fertility. Osteopontin is a glycoprotein found in several biological fluids including seminal plasma, and its presence is associated with spermatozoa concentration. In cattle, expression of osteopontin was highly correlated with bull fertility, and it was proposed to be a marker to predict male fertility (Cancel et al. 1999 Biol. Reprod. 60, 454–460). No data are available about the presence or activity of osteopontin in water buffalo. The aim of this preliminary study was to determine if osteopontin is present in buffalo semen and to evaluate whether freezing procedures cause the loss of osteopontin from spermatozoa. Semen was collected in authorized semen collection centers from 6 buffalo bulls by using an artificial vagina. A collection of bovine semen was used as a positive control. An aliquot from each sample was frozen using standard procedures for semen storage. Each ejaculate was centrifuged at 600g for 10 min at room temperature, and the supernatant was recovered and centrifuged at 10 000g for 1 h at 4�C. The total protein concentration in seminal plasma and spermatozoa was determined by the Bradford method, using ovoalbumin as the standard. Proteins (50 �g) were separated by electrophoresis and analyzed by western blotting (Cancel et al. 1999). Polyclonal antibodies against bovine milk osteopontin were prepared as previously described (Cancel et al. 1997 Biol. Reprod. 57, 1293–1301). The intensities of bands indicated by western blot were quantified by densitometer. Osteopontin was detected in all samples of buffalo semen. Most of the osteopontin detected was in the seminal plasma. Relative amounts of osteopontin detected in spermatozoa were 50% or less of that detected in seminal plasma; furthermore, the protein was not found in sperm from all bulls. These results suggest that most osteopontin is produced by the ampullae and seminal vesicles, similar to what was reported for cattle (Cancel et al. 1999). Semen frozen by standard procedures showed a reduction in amount of osteopontin by up to 50%. These studies suggest that the fertility-associated protein osteopontin may be useful as a sensitive tool to evaluate whether sperm storage procedures are detrimental and result in excess loss of osteopontin from sperm. In conclusion, the results have demonstrated that osteopontin is present in buffalo seminal plasma and sperm. Further studies will examine whether the expression of osteopontin is correlated with the fertility of buffalo bulls, as has been demonstrated in bovine bulls.