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T.L Yusuf
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In vitro embryo production through modification of time and gonadotropin hormone during oocytes maturation Purwantara, B; Diwyanto, K; Triwulaninngsih, Endang; Toelihere, M.R; Rutledge, J.J; Yusuf, T.L
Indonesian Journal of Animal and Veterinary Sciences Vol 6, No 3 (2001)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (221.222 KB) | DOI: 10.14334/jitv.v6i3.238

Abstract

This research has been conducted at the laboratory of in vitro fertilization of the University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. There is transportation constraint in importing oocytes from USA. It takes more than 24 hours to bring it to Indonesia. In fact, oocytes maturation and ready to be fertilized normally requires only 24 hours in 5% CO2 incubator at 38.5°C. Therefore, this research is needed to study the effect of gonadotropin hormone and time for oocyte maturity and ready to be fertilized at a period more than 24 hours. If this problem could be solved then the importation of oocytes could be cheaper and easier than importation of life animals or embryos. Ovaries were collected from slaughterhouse in Wisconsin. Oocytes were matured in TCM-199 medium in 5% CO2 incubator and at 30°C enriched with FSH 10 μl/ml, oestradiol 17 β 1μl/ml and 10 % FCS as control of gonadotropin hormone treatment (A); with FSH 10 μl/ml (B); with oestradiol 17 β 1μl/ml (C) and without gonadotropin hormone (D) for 24 hours, 30 hours and 36 hours as time of maturation treatment I, II and III respectively. The oocytes were fertilized in vitro with motile sperm selection by Percoll gradient and incubation between sperm and oocytes in fertilization media (TALP) for 20 hours. All zygotes were cultured in modification of KSOM medium up to blastocyst and were fed serum 5 μl/50 μl medium on day 6. Data were analyzed by SAS program. Percentage of cleavage between time of maturation were significant (p<0.01); between gonadotropin hormone treatment A vs B and A vs C and B vs C and B vs D and between A vs D were significant (p<0.01), but between treatment C vs D were not significant (p>0.05). Percentage of blastocyst between time of maturation were not significant (p>0.05), but between gonadotropin hormone treatment A vs B and A vs C and B vs D and C vs D were significant (p<0.01), but between treatment A vs D and B vs C were not significant (p>0.05). Percentage of cleavage, morula, blastocyst, expanded blastocyst and unertilized  ova on this study are 66.73%, 22.43%, 40.33%, 0.81% and 32.51 for 24 hours incubation (I); 61.55%, 25.69%, 32.69%,  0.54% and 27.61% for 30 hours incubation (II); 72.43%, 32.06%, 37.97%, 0.0% and 25.31% for 36 hours incubation (III) respectively. According to this study, in vitro production of embryo could be conducted at 30°C and incubation on maturation media for more than 24 hours.   Key words: Oocytes, cleavage, embryos, blastocyst
Using CR1aa versus KSOM as the culture medium for in vitro embryo production of cattle Triwulaninngsih, Endang; Toelihere, M.R; Rutledge, J.J; Yusuf, T.L; Purwantara, B; Diwyanto, K
Indonesian Journal of Animal and Veterinary Sciences Vol 7, No 1 (2002)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (237.936 KB) | DOI: 10.14334/jitv.v7i1.272

Abstract

This research has been conducted at the laboratory of in vitro fertilization in the Department of Animal Science University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. Oocytes were matured in TCM- 199 medium (in 5% CO2 incubator and at 390C) enriched with follicle stimulating hormone (FSH) 10 μl/ml, oestradiol 17 β 1μl/ml and 10% Fetal Calf Serum (FCS). The oocytes were fertilized in vitro with motile sperm and incubation between sperm and oocytes in fertilization medium Tyroide Albumin Lactate Pyruvate (TALP) for 20 hours. All zygotes were cultured in CR1aa (n=1549) medium versus modification of protein-free pottasium simplex optimized medium (KSOM) (n=675) up to blastocyst stage and were fed FCS 5 μl/50 μl medium on day 6, as treatment A and B respectively. Data were analyzed by completely randomized design with SAS program. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 91.4% vs 75.6 %; 75.6% vs 58.9%; 61.5% vs 38.5%; 31.2% vs 5.1%, 8.6% vs 24.4%, 15.7% vs 8% which were significantly different (P<0.01) for treatment CR1aa and KSOM respectively. Based on this study, CR1aa medium is better culture medium than KSOM for efficient in vitro production (IVP) of bovine embryos.   Key words: Oocytes, in vitro fertilization, embryo, blastocyst, culture medium
Quality of Garut ram frozen semen in various glycerol concentrations Rizal, Muhammad; Toelihere, M.R; Yusuf, T.L; Purwantara, B; Situmorang, P
Indonesian Journal of Animal and Veterinary Sciences Vol 7, No 3 (2002)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (144.127 KB) | DOI: 10.14334/jitv.v7i3.294

Abstract

Semen was collected once a week using artificial vagina from four mature Garut rams. Immediately after initial evaluation, semen was divided into three parts and diluted with tris extender containing 3% (G3), 5% (G5), and 7% (G7) glycerol, respectively, each with the concentration of 100 million motile sperm 0.25 ml-1. Semen was loaded in 0.25 ml mini straws, and equilibrated at 50C for three hours, then frozen and stored in liquid nitrogen container. Results indicated that percentages of post thawing motility and live sperm for G5 (40 and 50.50%) were significantly higher than G3 (32.50 and 45.33%) (P<0.05), but not significantly different with G7 (39.17 and 47.67%) (P>0.05). Percentages of post thawing intact acrosomal and plasma membrane for G5 (42.67 and 43.17%) were significantly higher than G3 (36.17 and 38.17%) (P<0.05), but not significantly different with G7 (38 and 39.83%) (P>0.05). In conclusion, concentration of 5% glycerol is the optimal dose in maintaining frozen semen quality of Garut rams.   Key words: Glycerol concentrations, frozen semen, Garut ram
Characteristics of reproductive performance of Garut rams Rizal, Muhammad; Toelihere, M.R; Yusuf, T.L; Purwantara, B; Situmorang, P
Indonesian Journal of Animal and Veterinary Sciences Vol 8, No 2 (2003)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (167.46 KB) | DOI: 10.14334/jitv.v8i2.383

Abstract

Basic information on reproductive potency of Garut rams is necessary in order to identify the capacity of rams in producing chilled or frozen semen. Eight Garut rams (three to five years old) were used in this study. The male sexual behaviors were observed and semen was collected once a week using artificial vagina. Semen quality was evaluated and its potency to produce frozen semen was calculated. Results of this study indicated that first, second, and third ejaculations were at the 29, 87 and 176th seconds, respectively. Fresh semen volume, sperm concentration, motility, intact acrosomal cap, and intact plasma membrane were 0.99 ml, 3224 million/ml; 76.67; 86.13 and 87.73%, respectively. Protein value, fructose, vitamin C, vitamin E, sodiu, potassium, calcium, magnesium, phosphor, chloride, and mangan in seminal plasma of fresh semen were 4140, 180, 3.2, 24, 180, 117, 9, 6.12, 60, 104, and 5 mg/ml, respectively. Measurement of head length, width, and length of sperm tail were 6.59, 3.99, and 42.65 μm, respectively. Length and width measurement of right and left testes, and scrotal circumference were 12.71, 6.5, and 32.36 cm, respectively. Capacity of each Garut rams to produce frozen semen from three consecutive ejaculations are 35.88 mini straw with the cencentration of 200 million motile sperm per 0.25 ml.   Key words: Reproductive characteristics, Garut rams  
The viability of fresh and extended semen of stallion with different sperm concentration in Dimitropoulos-modified extender ., Yudi; Arifiantini, I; Purwantara, B; Yusuf, T.L
Indonesian Journal of Animal and Veterinary Sciences Vol 13, No 1 (2008)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (80.697 KB) | DOI: 10.14334/jitv.v13i1.593

Abstract

The objective of the experiment was to study the motility and viability of spermatozoa of fresh semen, and the quality of extended semen with different sperm concentration in Dimitropoulos-modified extender. Semen was collected using artificial vagina from three 4-8 year old stallions (different breed). Semen characteristics and quality was evaluated macro- and microscopically. For longevity evaluation, semen was stored at room and chilled temperature, and was evaluated for motility and viability every 3 h. Prior extension, semen was centrifugated at 3000 rpm for 20 minutes. The condensed sperm was re-suspended in Dimitropoulos (DV) supplemented with 50 mM fructose with the concentration of 200, 100 and 50 x 106 spz/mL. All samples were stored at room and chilled temperature, and was evaluated for motility and viability every 3 h and 12 h for room and chilled temperature. Results of the experiments indicated that fresh semen characteristics was fairly good. For longevity evaluation, semen with motility of 48.33 and 10.42% was observed at 3 h and 12 h after the onset of storage. The extended-semen with 50 x 106 spz/mL showed significantly higher in term of motility and viability (P<.05) than that with 200 x 106 spz/mL, but for that of 100 x 106 spz/mL. It is recommended that sperm concentration should be 50 x 106 spz/ml for a long period storage with reasonable good quality. Key words: Stallion, Semen, Sperm Concentration, Dimitropoulos
In vitro embryo production through modification of time and gonadotropin hormone during oocytes maturation Endang Triwulaninngsih; M.R Toelihere; J.J Rutledge; T.L Yusuf; B Purwantara; K Diwyanto
Jurnal Ilmu Ternak dan Veteriner Vol 6, No 3 (2001): SEPTEMBER 2001
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (221.222 KB) | DOI: 10.14334/jitv.v6i3.238

Abstract

This research has been conducted at the laboratory of in vitro fertilization of the University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. There is transportation constraint in importing oocytes from USA. It takes more than 24 hours to bring it to Indonesia. In fact, oocytes maturation and ready to be fertilized normally requires only 24 hours in 5% CO2 incubator at 38.5°C. Therefore, this research is needed to study the effect of gonadotropin hormone and time for oocyte maturity and ready to be fertilized at a period more than 24 hours. If this problem could be solved then the importation of oocytes could be cheaper and easier than importation of life animals or embryos. Ovaries were collected from slaughterhouse in Wisconsin. Oocytes were matured in TCM-199 medium in 5% CO2 incubator and at 30°C enriched with FSH 10 μl/ml, oestradiol 17 β 1μl/ml and 10 % FCS as control of gonadotropin hormone treatment (A); with FSH 10 μl/ml (B); with oestradiol 17 β 1μl/ml (C) and without gonadotropin hormone (D) for 24 hours, 30 hours and 36 hours as time of maturation treatment I, II and III respectively. The oocytes were fertilized in vitro with motile sperm selection by Percoll gradient and incubation between sperm and oocytes in fertilization media (TALP) for 20 hours. All zygotes were cultured in modification of KSOM medium up to blastocyst and were fed serum 5 μl/50 μl medium on day 6. Data were analyzed by SAS program. Percentage of cleavage between time of maturation were significant (p<0.01); between gonadotropin hormone treatment A vs B and A vs C and B vs C and B vs D and between A vs D were significant (p<0.01), but between treatment C vs D were not significant (p>0.05). Percentage of blastocyst between time of maturation were not significant (p>0.05), but between gonadotropin hormone treatment A vs B and A vs C and B vs D and C vs D were significant (p<0.01), but between treatment A vs D and B vs C were not significant (p>0.05). Percentage of cleavage, morula, blastocyst, expanded blastocyst and unertilized  ova on this study are 66.73%, 22.43%, 40.33%, 0.81% and 32.51 for 24 hours incubation (I); 61.55%, 25.69%, 32.69%,  0.54% and 27.61% for 30 hours incubation (II); 72.43%, 32.06%, 37.97%, 0.0% and 25.31% for 36 hours incubation (III) respectively. According to this study, in vitro production of embryo could be conducted at 30°C and incubation on maturation media for more than 24 hours.   Key words: Oocytes, cleavage, embryos, blastocyst
Quality of Garut ram frozen semen in various glycerol concentrations Muhammad Rizal; M.R Toelihere; T.L Yusuf; B Purwantara; P Situmorang
Jurnal Ilmu Ternak dan Veteriner Vol 7, No 3 (2002): SEPTEMBER 2002
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (144.127 KB) | DOI: 10.14334/jitv.v7i3.294

Abstract

Semen was collected once a week using artificial vagina from four mature Garut rams. Immediately after initial evaluation, semen was divided into three parts and diluted with tris extender containing 3% (G3), 5% (G5), and 7% (G7) glycerol, respectively, each with the concentration of 100 million motile sperm 0.25 ml-1. Semen was loaded in 0.25 ml mini straws, and equilibrated at 50C for three hours, then frozen and stored in liquid nitrogen container. Results indicated that percentages of post thawing motility and live sperm for G5 (40 and 50.50%) were significantly higher than G3 (32.50 and 45.33%) (P<0.05), but not significantly different with G7 (39.17 and 47.67%) (P>0.05). Percentages of post thawing intact acrosomal and plasma membrane for G5 (42.67 and 43.17%) were significantly higher than G3 (36.17 and 38.17%) (P<0.05), but not significantly different with G7 (38 and 39.83%) (P>0.05). In conclusion, concentration of 5% glycerol is the optimal dose in maintaining frozen semen quality of Garut rams.   Key words: Glycerol concentrations, frozen semen, Garut ram
The viability of fresh and extended semen of stallion with different sperm concentration in Dimitropoulos-modified extender Yudi .; I Arifiantini; B Purwantara; T.L Yusuf
Jurnal Ilmu Ternak dan Veteriner Vol 13, No 1 (2008): MARCH 2008
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (80.697 KB) | DOI: 10.14334/jitv.v13i1.593

Abstract

The objective of the experiment was to study the motility and viability of spermatozoa of fresh semen, and the quality of extended semen with different sperm concentration in Dimitropoulos-modified extender. Semen was collected using artificial vagina from three 4-8 year old stallions (different breed). Semen characteristics and quality was evaluated macro- and microscopically. For longevity evaluation, semen was stored at room and chilled temperature, and was evaluated for motility and viability every 3 h. Prior extension, semen was centrifugated at 3000 rpm for 20 minutes. The condensed sperm was re-suspended in Dimitropoulos (DV) supplemented with 50 mM fructose with the concentration of 200, 100 and 50 x 106 spz/mL. All samples were stored at room and chilled temperature, and was evaluated for motility and viability every 3 h and 12 h for room and chilled temperature. Results of the experiments indicated that fresh semen characteristics was fairly good. For longevity evaluation, semen with motility of 48.33 and 10.42% was observed at 3 h and 12 h after the onset of storage. The extended-semen with 50 x 106 spz/mL showed significantly higher in term of motility and viability (P<.05) than that with 200 x 106 spz/mL, but for that of 100 x 106 spz/mL. It is recommended that sperm concentration should be 50 x 106 spz/ml for a long period storage with reasonable good quality. Key words: Stallion, Semen, Sperm Concentration, Dimitropoulos
Using CR1aa versus KSOM as the culture medium for in vitro embryo production of cattle Endang Triwulanningsih; M.R Toelihere; J.J Rutledge; T.L Yusuf; B Purwantara; K Diwyanto
Jurnal Ilmu Ternak dan Veteriner Vol 7, No 1 (2002): MARCH 2002
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (237.936 KB) | DOI: 10.14334/jitv.v7i1.272

Abstract

This research has been conducted at the laboratory of in vitro fertilization in the Department of Animal Science University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. Oocytes were matured in TCM- 199 medium (in 5% CO2 incubator and at 390C) enriched with follicle stimulating hormone (FSH) 10 μl/ml, oestradiol 17 β 1μl/ml and 10% Fetal Calf Serum (FCS). The oocytes were fertilized in vitro with motile sperm and incubation between sperm and oocytes in fertilization medium Tyroide Albumin Lactate Pyruvate (TALP) for 20 hours. All zygotes were cultured in CR1aa (n=1549) medium versus modification of protein-free pottasium simplex optimized medium (KSOM) (n=675) up to blastocyst stage and were fed FCS 5 μl/50 μl medium on day 6, as treatment A and B respectively. Data were analyzed by completely randomized design with SAS program. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 91.4% vs 75.6 %; 75.6% vs 58.9%; 61.5% vs 38.5%; 31.2% vs 5.1%, 8.6% vs 24.4%, 15.7% vs 8% which were significantly different (P<0.01) for treatment CR1aa and KSOM respectively. Based on this study, CR1aa medium is better culture medium than KSOM for efficient in vitro production (IVP) of bovine embryos. Key words: Oocytes, in vitro fertilization, embryo, blastocyst, culture medium
Characteristics of reproductive performance of Garut rams Muhammad Rizal; M.R Toelihere; T.L Yusuf; B Purwantara; P Situmorang
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 2 (2003): JUNE 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (167.46 KB) | DOI: 10.14334/jitv.v8i2.383

Abstract

Basic information on reproductive potency of Garut rams is necessary in order to identify the capacity of rams in producing chilled or frozen semen. Eight Garut rams (three to five years old) were used in this study. The male sexual behaviors were observed and semen was collected once a week using artificial vagina. Semen quality was evaluated and its potency to produce frozen semen was calculated. Results of this study indicated that first, second, and third ejaculations were at the 29, 87 and 176th seconds, respectively. Fresh semen volume, sperm concentration, motility, intact acrosomal cap, and intact plasma membrane were 0.99 ml, 3224 million/ml; 76.67; 86.13 and 87.73%, respectively. Protein value, fructose, vitamin C, vitamin E, sodiu, potassium, calcium, magnesium, phosphor, chloride, and mangan in seminal plasma of fresh semen were 4140, 180, 3.2, 24, 180, 117, 9, 6.12, 60, 104, and 5 mg/ml, respectively. Measurement of head length, width, and length of sperm tail were 6.59, 3.99, and 42.65 μm, respectively. Length and width measurement of right and left testes, and scrotal circumference were 12.71, 6.5, and 32.36 cm, respectively. Capacity of each Garut rams to produce frozen semen from three consecutive ejaculations are 35.88 mini straw with the cencentration of 200 million motile sperm per 0.25 ml.   Key words: Reproductive characteristics, Garut rams  
Co-Authors Anak Agung Gede Sugianthara B Purwantara B Purwantara Endang Triwulaninngsih Endang Triwulanningsih I Arifiantini J.J Rutledge K Diwyanto M.R Toelihere MUHAMMAD RIZAL P Situmorang Yudi .