TAKDIR SAILI
Bagian Klinik Hewan, Fakultas Kedokteran Hewan, Universitas Udayana, Bali

Published : 38 Documents
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Sperm Preservation using Freeze-Drying Method

HAYATI Journal of Biosciences Vol 12, No 1 (2005): March 2005
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI) for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF) the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.

INJEKSI SPERMATOZOA DOMBA HASIL PENGERINGBEKUAN KE DALAM SEL TELUR MENGGUNAKAN TEKNIK INTRACYTOPLASMIC SPERM INJECTION (ICSI) INJECTION OF FREEZE-DRIED RAM SPERMATOZOA INTO OOCYTES USING INTRACYTOPLASMIC SPERM INJECTION, ICSI

Jurnal Veteriner Vol 8, No 1 (2007)
Publisher : Jurnal Veteriner

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Abstract

Pada penelitian ini dikaji kemampuan spermatozoa domba hasil pengeringbekuan untuk melakukan dekondensasi dan membentuk pronukleus setelah diinjeksikan ke dalam oosit dengan menggunakan teknik intracytoplasmic sperm injection (ICSI). Metode pewarnaan aceto lacmoid digunakan untuk mengevaluasi kejadian dekondensasi dan pembentukan pronukleus pada oosit setelah ICSI. Hasil penelitian menunjukkan bahwa spermatozoa hasil pengeringbekuan dapat melakukan dekondensasi (2%) dan mendukung pembentukan 1PN (34%) tetapi belum mampu mendukung pembentukan 2PN setelah diinjeksikan ke dalam oosit. Sedangkan injeksi dengan menggunakan spermatozoa segar mampu mendukung pembentukan 2PN (30%) dan 1PN (40%). Sebagai kesimpulan dapat dikemukakan bahwa spermatozoa hasil pengeringbekuan mampu melakukan dekondensasi dan mendukung pembentukan 1PN setelah ICSI

PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN

Jurnal Veteriner Vol 10, No 4 (2009)
Publisher : Jurnal Veteriner

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Abstract

Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.

PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN

Jurnal Veteriner Vol 10, No 4 (2009)
Publisher : Jurnal Veteriner

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Abstract

Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.

Aktivasi Oosit Menggunakan Strontium Klorida setelah Injeksi dengan Spermatozoa Domba Hasil Pengeringbekuan (OOCYTE ACTIVATION USING STRONTIUM CHLORIDE FOLLOWING INJECTION OF FREEZE-DRIED RAM SPERMATOZOA)

Jurnal Veteriner Vol 13, No 3 (2012)
Publisher : Jurnal Veteriner

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Abstract

One of the factors that inhibit the formation of male pronuclei following injection of freeze-dried ramspermatozoa was the absence of artificial activation during oocyte incubation after the injection. Therefore,in this experiment the ability of strontium chloride (SrCl2) to improve oocyte activation followingintracytoplasmic sperm injection (ICSI) was evaluated. Aceto lacmoid staining was used to assessdecondensation and pronucleus formation following ICSI. Results of this experiment revealed that freezedriedspermatozoa had the ability to decondense and to form 1PN following injection into oocytes evenwithout artificial activation, but failed to form 2PN. However, 40% of 2PN oocytes were obtained when theinjected oocytes was first incubated for 20 minutes in medium containing 50 mM strontium chloride thensubsequently incubated for 10 hours in medium without strontium. On the contrary, the 2PN oocytes werenot observed either in injected oocyte neither without artificial activation nor in non-injected oocytes withartificial activation. In conclusion, freeze-dried ram spermatozoa were able to decondense and to support2PN formation following ICSI and artificial activation using strontium.

Freeze-Drying Spermatozoa as an Alternative Method for Rescuing Genetic Material of Animal

WARTAZOA. Indonesian Bulletin of Animal and Veterinary Sciences Vol 18, No 4 (2008): DECEMBER 2008
Publisher : Indonesian Center for Animal Research and Development

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Abstract

Cryopreservation is one of the commonly methods used in spermatozoa preservation in which sperm is frozen and stored in the container of liquid nitrogen. The frozen sperm is still motile after thawing, so it is possible to use it in both artificial insemination and in vitro fertilization to produce an embryo. However, this technique needs a continuous supply of liquid nitrogen and a container as a place to store the frozen sperm. The advanced technique in microinjection has led the possibility of using immotile sperm to fertilize oocyte. Therefore, the sperm preservation method may be simplified because the motility of sperm has not been taken into consideration in fertilization compared to the previous method. Freeze-drying sperm is the proposed method in which the sperm is frozen and sublimated using freeze-drying machine to produce freeze-dried sperm. The freeze-dried sperm might be stored in room temperature or in refrigerator. Several reports have claimed that freeze-dried sperm is not motile but it still has capability to fertilize oocyte, even produces offspring, because its DNA remains intact.   Key words: Cryopreservation, freeze-drying, sperm, oocyte

Status of ram spermatozoa DNA after freeze-drying process

Jurnal Ilmu Ternak dan Veteriner Vol 11, No 3 (2006): SEPTEMBER 2006
Publisher : Indonesian Center for Animal Research and Development (ICARD)

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Abstract

The process of freeze drying caused detrimental effect on plasma membrane and acrosome of the spermatozoa, even it potentially could alter the chromatin and DNA integrities. On the other hand, DNA integrity is essential for spermatozoa to participate in pronucleus formation during fertilization event. Therefore the evaluation of DNA integrity should be carried out to study the effect of freeze drying process. EDTA, EGTA, and PBS were used as dilution media of spermatozoa prior to freeze drying process to protect the DNA. Toluidine blue staining and comet assay methods were used to evaluate the alteration on chromatin and DNA integrities of spermatozoa, respectively. The results revealed that the highest compacted chromatin after 6 months storage of freeze-dried spermatozoa were observed from EGTA-3 (98%) and EGTA-1 (97%) treatments that had significant differences compared to all PBS treatments (90-92%), but not for fresh spermatozoa (100%). Whereas, the highest compacted DNA integrity of freeze-dried spermatozoa were observed from EGTA-2 (92%) and EGTA-3 (92%) but had no significant differences compared to other treatments including fresh spermatozoa (97%). These results demonstrate that EDTA and EGTA tend to be able to protect chromatin and DNA integrities of ram spermatozoa during freeze-drying and storage compared to PBS. Key Words: Freeze-Drying, Spermatozoa, DNA, Toluidine Blue, Comet Assay

IMBANGAN PROTEIN DAN ENERGI BERBEDA DALAM RANSUM PUYUH FASE GROWER TERHADAP KONSUMSI PAKAN, PERTAMBAHAN BOBOT BADAN, DAN KONVERSI RANSUM

Jurnal Ilmu dan Teknologi Peternakan Tropis Vol 5, No 2 (2018): JITRO, Mei
Publisher : Jurnal Ilmu dan Teknologi Peternakan Tropis

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Abstract

ABSTRAK             Kandungan energi dan protein pakan merupakan faktor yang mempengaruhi kualitas pakan dan performans produksi ternak. Penelitian ini bertujuan untuk mempelajari pengaruh imbangan energi dan protein berbeda dalam ransum puyuh fase grower terhadap konsumsi pakan, perrtambahan bobot badan, dan konversi pakan. Seratus dua puluh DOQ disebar secara acak pada 24 unit kandang percobaan. Perlakuan yang dicobakan terdiri atas 2 level energi pakan (2700 dan 2900 kkal/kg) dan 3 level protein pakan (18, 20, dan 22%), sehingga terdapat 6 kombinasi perlakuan, yaitu R1 (2700 EM – 18% PK), R2 (2700 EM – 20% PK), R3 (2700 EM – 22% PK), R4 (2900 EM – 18% PK), R5 (2900 EM – 20% PK), dan R6 (2900 EM – 22% PK). Pakan yang dicobakan merupakan pakan self mixing. Parameter yang diamati adalah konsumi pakan, pertambahan bobot badan, dan konversi pakan puyuh umur 2 hingga 6 minggu. Data yang diperoleh dianalisis menggunakan analisis ragam dan dilanjutkan dengan uji wilayah berganda duncan. Hasil penelitian menunjukkan bahwa imbangan energi-protein pakan berbeda tidak memberikan pengaruh (P>0,05) pada konsumsi pakan, pertambahan bobot badan, dan konversi pakan. Kombinasi energi metabolisme 2700 kkal/kg dan 18% protein sudah dapat memenuhi kebutuhan nutrisi puyuh periode grower.Kata Kunci: protein, energi, puyuh, grower   ABSTRACT             Energy and protein that contained in poultry feed is a factor that affect the feed quality and poultry production performance. This research aimed to study the effect of different energy and protein balance in quail feed on feed consumption, weight gain, and feed conversion ratio. One hundred and twenty day old quails were divided into 24 units enclosure research. The trial feed was consist of 2 levels of energy feed (2700 and 2900 kcal/kg) and 3 levels of crude protein (18, 20, and 22% CP), so that there were 6 combinations of treatments, i.e. R1 (2700 ME – 18% CP), R2 (2700 ME – 20% CP), R3 (2700 ME – 22% CP), R4 (2900 ME – 18% CP), R5 (2900 ME – 20% CP), R6 (2900 ME – 22% CP). The used feed was a self mixing feed. The observed parameters were feed consumption, body weight gain, and feed conversion ratio of quail at 2-6 weeks of age. The data obtained were analyzed using variance analysis and continued using Duncan’s multiple range test. The result showed that the balance of energy-protein in quail feed did not affect (P>0,05) feed consumption, body weight gain, and feed conversion ratio. The combination of 2700 kcal/kg metabolizable energy and 18% cruse protein could already maintain the needs of the grower period of quail nutrients.Keywords: protein, energy, quail, grower

Freeze-Drying Spermatozoa as an Alternative Method for Rescuing Genetic Material of Animal

Indonesian Bulletin of Animal and Veterinary Sciences Vol 18, No 4 (2008)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Original Source | Check in Google Scholar | Full PDF (96.992 KB)

Abstract

Cryopreservation is one of the commonly methods used in spermatozoa preservation in which sperm is frozen and stored in the container of liquid nitrogen. The frozen sperm is still motile after thawing, so it is possible to use it in both artificial insemination and in vitro fertilization to produce an embryo. However, this technique needs a continuous supply of liquid nitrogen and a container as a place to store the frozen sperm. The advanced technique in microinjection has led the possibility of using immotile sperm to fertilize oocyte. Therefore, the sperm preservation method may be simplified because the motility of sperm has not been taken into consideration in fertilization compared to the previous method. Freeze-drying sperm is the proposed method in which the sperm is frozen and sublimated using freeze-drying machine to produce freeze-dried sperm. The freeze-dried sperm might be stored in room temperature or in refrigerator. Several reports have claimed that freeze-dried sperm is not motile but it still has capability to fertilize oocyte, even produces offspring, because its DNA remains intact.   Key words: Cryopreservation, freeze-drying, sperm, oocyte

Status of ram spermatozoa DNA after freeze-drying process

Indonesian Journal of Animal and Veterinary Sciences Vol 11, No 3 (2006)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Original Source | Check in Google Scholar

Abstract

The process of freeze drying caused detrimental effect on plasma membrane and acrosome of the spermatozoa, even it potentially could alter the chromatin and DNA integrities. On the other hand, DNA integrity is essential for spermatozoa to participate in pronucleus formation during fertilization event. Therefore the evaluation of DNA integrity should be carried out to study the effect of freeze drying process. EDTA, EGTA, and PBS were used as dilution media of spermatozoa prior to freeze drying process to protect the DNA. Toluidine blue staining and comet assay methods were used to evaluate the alteration on chromatin and DNA integrities of spermatozoa, respectively. The results revealed that the highest compacted chromatin after 6 months storage of freeze-dried spermatozoa were observed from EGTA-3 (98%) and EGTA-1 (97%) treatments that had significant differences compared to all PBS treatments (90-92%), but not for fresh spermatozoa (100%). Whereas, the highest compacted DNA integrity of freeze-dried spermatozoa were observed from EGTA-2 (92%) and EGTA-3 (92%) but had no significant differences compared to other treatments including fresh spermatozoa (97%). These results demonstrate that EDTA and EGTA tend to be able to protect chromatin and DNA integrities of ram spermatozoa during freeze-drying and storage compared to PBS. Key Words: Freeze-Drying, Spermatozoa, DNA, Toluidine Blue, Comet Assay