Mohamad Agus Setiadi
Bagian Reproduksi dan Kebidanan Departemen Klinik Reproduksi dan Patologi Fakultas Kedokteran Hewan Institut Pertanian Bogor, Bogor

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Karakteristik dan komposisi semen kancil (Tragulus javanicus) yang dikoleksi dengan elektroejakulator

Jurnal Anatomi Indonesia Vol 1, No 1 (2006)
Publisher : Jurnal Anatomi Indonesia

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Abstract

The ejaculates were taken by electroejaculation from four apparently healthy young adultmale lessermousedeer (Tragulus javanicus). The animals were anesthetized with a combination of xylazin and ketamine followedby ether per inhalation anesthesia. The semen was white, yellowish, or creamy in color. The semen had a pH of7.63±0.22. Themean values for volumewas 19.44±6.8 μl, spermconcentration was 47.44 ± 4.9 x 106 sperm/ml,percentage of sperm motility was 36.43±1.1 %, percentage of live sperm was 53.11±3.0 %, percentage ofabnormal spermatozoa was 21.03±1.05%and percentage of intact acrosome was 52.28±2.7%, respectively.These values were relatively lowwhen compared to other domestic ruminants and suggested to be relatedwithage and sexualmaturity of the animal.The seminal plasma contained 10.2–11.5mg/100ml fructose, 22.07–24.5mg/100ml citric acid, 65mg/100ml proteins, 22.07–24.5mg/100ml sorbitol, 91.1– 94.72mg/100ml sodium, 0.1mg/100ml potassium, 12.8mg/100ml calcium, 0.8mg/100mlmagnesiumand 10.72–11.2 chloride. By SDS PAGE,eleven proteins with different molecular weight were determined in the seminal plasma. Among them, theproteins with 64.77 kDa and 71.72 kDa were particularly prominent.

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.

Maturation Rate of Ovine Oocytes from Different Reproductive Status and Maturation Medium

HAYATI Journal of Biosciences Vol 13, No 4 (2006): December 2006
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The aim of the present study was to determine the number of follicles, oocyte quality and maturation rate of oocytes from pairs of ovary with different reproductive status in two maturation medium, TCM-199 as control and CR1aa as treatment. The pairs of ovary were classified into four groups: (i) ovaries with corpus luteum (CL) and dominant follicle (DF), (ii) ovaries with CL, without DF, (iii) ovaries with DF, without CL, (iv) ovaries without both CL and DF. Results of the experiment revealed that the greatest number of follicles was observed from ovary with CL without DF (15.88 + 10.68), although not significantly different (P > 0.05) with other status of ovaries. The lowest number (P < 0.05) of A grade oocytes was found from ovary with DF without CL (1.20 + 1.10). The percentage of Metaphase II was highest in TCM-199 (75.51%) with oocytes from ovaries with CL and DF, and the lowest with oocytes from ovaries with DF without CL in TCM-199 and CR1aa (42.86 and 30.95%). The study suggested that the number of oocytes with A grade were influenced by the reproductive status of ovaries. The maturation rate of A grade oocytes was influenced by the quality of oocytes and the composition of maturation medium. Key words: reproductive status, corpus luteum, dominant follicles, TCM-199, CR1aa

Koleksi Sel Telur dengan Teknik Laparoskopi untuk Produksi Embrio dan Transfer Embrio pada Domba

Jurnal Ilmu Pertanian Indonesia Vol 12, No 2 (2007): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

An experiment was carried out to analyze the application of laparoscopic technique for oocyte collection, in vitro embryo production and embryo transfer in sheep. The first experiment was conducted to observe effect of gonadotropin stimulation on follicle development and laparoscopic technique for oocytes aspiration. In the second experiment, effect of culture system on the embryo development in vitro was assessed and in the third experiment, the application of laparoscopic for embryo transfer has been conducted. The result showed that single dose of gonadotrophin was sufficient to support follicle development significantly and it could help follicle visualization. It also showed that laparoscopic ovum-pick up could be conducted weekly without any restriction The second series experiment showed CR1aa culture system was better than TCM 199 (29.90°/o vs 8.00%) and the changing of media was required to ensure better metabolism process for embryos. The third experiment revealed that embryo transfer could be conducted with an aid from laparoscope. In conclusion, single dose PMSG stimulation is sufficient to support follicle development for /aparoscopic ovum-pick up, the culture media changing affects the successful rate of in vitro embryo production (8% vs 25.66%) and the laparoscopy technique can be used safely for embryo transfer on sheep.Keyword: laparoscopic, oocyte, embryo transfer, sheep

Follicular dynamic and repeatability of follicular wave development in Peranakan Ongole (PO) cattle

Jurnal Ilmu Ternak dan Veteriner Vol 21, No 1 (2016): MARCH 2016
Publisher : Indonesian Center for Animal Research and Development (ICARD)

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Abstract

Superovulation treatment on PO cattle (Bos indicus) was less responsive compared to Bos taurus breed. It might due to the difference of their follicular dynamic. This study was conducted to investigate the follicular dynamics and its repeatability in PO cattle. Follicular dynamics observations conducted on 9 cows trough ultrasound scanning every day. Observations of wave patterns repeatability were performed in 6 cows which its wave pattern already known on the next consecutive IOI.  Research result indicated that PO cattle had 3 (66%) and 4-waves (34%) pattern. The first wave of 3 and 4-waves pattern emerged on day -0.4+0.9 and 1.4+1.1 respectively.  The second wave of 3 and 4-wave pattern emerged on day 9.8+1.5 and 7.4+1.9 respectively.  The pattern of 3 waves has a longer follicle dominant duration (11.6+1.5 day) in the first wave of estrous cycle, compared with 4 waves pattern (10+2.92 and 7+1.00 day respectively). The growth rate of dominant follicle was not different significantly between the 3 and 4-waves pattern (0.87+0.23 and 0.94+0.25 mm/day respectively). Similarly, ovulatory follicle diameter between 3 and 4-waves pattern was also not different significantly (12.24+12.34 and 12.30+12.23 mm respectively). Observation of wave patterns repeatability in 6 PO cows indicated that PO cattle had high repeatability in follicular wave pattern (0.88) and the number of growing follicle was 0.91.  This study resulted data for dynamic of follicular development, wave pattern, its repeatability which be expected to design the protocol of superovulation treatment or other reproduction technologies based on follicular dynamic to improve its result in PO cattle. 

Dinamika Ovarium Selama Siklus Estrus pada Domba Garut (OVARIAN DYNAMIC DURING THE ESTROUS CYCLE IN GARUT EWES)

Jurnal Veteriner Vol 13, No 2 (2012)
Publisher : Jurnal Veteriner

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Abstract

Ovarian dynamics in the garut ewes had never been studied continuously by using ultrasonography.The aim of this study was to observe development of the follicles and corpus luteums in the estrous cyclein the garut ewes. Garut ewes (n=6) with body weight 30.00±4.05 kg which had normal estrous cycle wereused in this study. All ewes were synchronized by using CIDR-G implantation for 14 days. Ovulation of thedominant follicle, development of the follicle waves and corpus luteum were observed continuously everyday during the estrous cycle after CIDR-G removal. The number of small (2-3 mm in diameter), medium (4-5 mm in diameter) and large (>5 mm in diameter) follicles were aligned during the estrous cycle. Follicleand corpus luteum diameters were measured by using built in caliper in the ultrasound. The resultsshowed 1) the average length of estrous cycle was 19,2±0,8 days; 2) ovarian follicle growth occurred inthree waves during the estrous cycle; 3) the number of preovulatory follicles were 1-2 follicles; 4) theaverage maximum diameters of preovulatory follicle was 7.5±0.5 mm; 5) the average maximum diametersof corpus luteum was 7.3±0.4 mm. In conclusion, the estrous cycle in garut ewes was 18-20 days with 3follicular waves.

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.

Tingkat Pematangan Inti Oosit Domba dan Pembentukan Pronukleus Setelah Parthenogenesis dengan Penambahan Glutathione (NUCLEAR MATURATION RATE OF OVINE OOCYTES AND PRONUCLEAR FORMATION AFTER PARTHENOGENESIS WITH GLUTATHIONE ADDITION)

Jurnal Veteriner Vol 13, No 4 (2012)
Publisher : Jurnal Veteriner

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Abstract

The aim of this study was to investigate the nuclear maturation rate of ovine oocytes and pronuclearformation following parthenogenesis with glutathione (GSH) addition in maturation and culture medium.In the first experiment, acolytes were matured in tissue culture medium (TCM) 199 with 0 (control), 0.25,0.5 and 1 mM glutathione (GSH) addition. In the second experiment, oocytes were matured in maturationmedium, then parthenogenetically activated by exposing to 7% ethanol (v/v) for 7 min, followed by treatmentwith 5 ìg/ml cytochalasin B for 4 h. Oocytes then cultured in medium TCM 199 + 10% FBS with treatmentswithout addition of 1 mM GSH (T0), addition in maturation medium (T1), addition in culture medium (T2),and addition in both maturation and culture medium (T3) then incubated at 38,5oC with 5% CO2 for 20-24h. The results showed that, nuclear maturation rate was not significantly different (P>0.05) among fourtreatments. The percentage of oocytes reached metaphases II (MII) stage were 79.71%, 79.07%, 80.95%and 84.13%, respectively. Percentages of activated oocyte with T1 (65.31%) and T3 (67.27%) were higher(P<0.01) compared to T0 (46.81%) and T2 (54.35%). However, T3 was not significantly different with T1. Inconclusion, the addition of GSH in maturation medium could not improve nuclear maturation rate butmore effective in supporting the number of activated oocytes.

Intracytoplasmic Sperm Injection (Icsi) Sebagai Teknik Reproduksi Bantuan Unggulan

Jurnal Sain Veteriner Vol 23, No 1 (2005)
Publisher : Fakultas Kedokteran Hewan

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Abstract

Beberapa teknik fertilisasi mikro telah diterapkan untuk mengatasi masalah infertilitas pria, namun hanya intracytoplasmic sperm injection (ICSI) yang mampu meningkatkan secara nyata keberhasilan fertilisasi, implantasi dan kelahiran. Pada perkembangannya teknik ICSI juga telah diterapkan pada beberapa jenis hewan sebagai suatu model untuk mempelajari kemampuan fertilisasi berbagai jenis spermatozoa yang secara alami atau melalui fertilisasi in vitro tidal( bisa membuahi sel telur. Walaupun keberhasilan teknik ICSI telah dapat mengatasi masalah infertilitas pada pria, teknik ICSI masih mempunyai potensi negatif yang mungkin muncul pada anak hasil ICSI. Namun demikian, hal ini perlu penelitian lebih lanjut untuk membuktikannya.

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