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The effects of proline, carnitine on the viability of sperm stored at 5oC (chilled semen) Situmorang, Polmer; Triwulanningsih, E; Lubis, A; Caroline, W; Sugiarti, T
Indonesian Journal of Animal and Veterinary Sciences Vol 6, No 1 (2001)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (147.85 KB) | DOI: 10.14334/jitv.v6i1.211

Abstract

An experiment was carried out to evaluate the addition of proline, carnitine in Tris-extender on the viability of bull sperm following storage at 5oC. Semen was collected by means of artificial vagina (AV), diluted in Tris-extender containing 5% V/V egg yolk (EY) and 4% V/V glycerol to get a final concentration of 50 million sperms/ml. Diluted semen cooled to 5oC for 45 minute and stored at those temperature for 1, 3, 10, and 13 days. In the first activity the addition of 15, 30 and 60 mM carnitine in Tris-extender while in the second activity the inclusion of 15, 30, and 60 mM proline on the viability of sperm was investigated. Addition of carnitine to Tris-extender signifinatly increase (P<0.05) the viability of sperm after storage for more than 3 days. At 3 days of storage, the mean %M and %L were 27.3, 38.8, 33.5, 53.0, 31.8, 47.0, and 30.5, 46.8 for control 15, 30, and 60 mM carnitine respectively. The similar results was obtained for 7 days of storage where the mean %M and %L for control (12.5 and 27.3) was significantly lower (P<0.05) than those 15, 30, and 60 mM carnitine (15.0, 33.5, 18.8, 36.5, 17.5, 36.3). The superiority of carnitine was maintained for 10 days of storage, where the mean %L were 23.5, 28.8, 31.5, and 30.3 for control; 15; 30; and 60 mM respectively. There was no any significant within concentration of carnitine tested (15 to 60 mM).The condition of apical ridge was not significantly affected by carnitine. In the second activity, inclusion of proline to Trisextender statistically (P<0.05) improved the viability of sperm after storage for 7 and 13 days. After 7 days of storage the mean %M and %L were 31.4, 36.4, 38.8, 40.4, 36.6, 42.7, and 34.8, 43.3 for control; 15, 30, and 60 mM proline respectively. The significant effects of proline was remain for 13 days of storage where the mean %M and % L were 24.6, 32.9, 28.6, 37.5, 29.1, 39.8, and 30.1, 37.3 for control; 15, 30, and 60 mM proline respectively. There was no significant difference within the concentration of proline. Condition of apical ridge was not significantly affected by proline.   Key words: Sperm, viability, carnitine, proline
Effect of exogenous glutathione in medium fertilization to improve blastocyst rates of bovine embryos Triwulaninngsih, E; Situmorang, P; Putu, I-G; Sugiarti, T; Lubis, A.M; Kusumaningrum, D.A; Caroline, W; Sianturi, R.G
Indonesian Journal of Animal and Veterinary Sciences Vol 7, No 2 (2002)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (190.218 KB) | DOI: 10.14334/jitv.v7i2.283

Abstract

Glutathione (C10H17N3O6S) is a tripeptide (γ-Glu-Cys-Gly) widespread in living organism. Glutathione (GSH) at the 5 mM concentration increased the motility and fertility of frozen-thawed sperm. Intracellulair glutathione improved the cleavage rate and embryo development to the blastocyst rate. Research on in vitro embryos production through the implementation of GSH during IVF (in vitro fertilization) on embryo development has been conducted at the Laboratorium Reproductive of Physiology, Research Institute for Animal Production. Ovaries of beef cattle as source of oocytes were collected from the slaughterhouse in a thermo flask with 350C PBS as medium and transported to the laboratory. The oocytes were fertilized in vitro with selected motile sperm using Percoll gradient (90 and 45%). Ten COCs (cumulus oocytes complexes) were transfered to 44 μl of fertilization medium (mTALP) was performed with either 0; 0.25; 0.50; 0.75 and 1.00 mM of glutathione as treatment A, B, C, D and E respectively, and inseminated with 2 μl of capacitated sperm and added 2 μl of heparin and 2 μl of PHE (consisting of 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine). Incubation between sperm and oocytes in the 5% CO2 incubator and RH 90% in fertilization media (TALP) for 20 hours. All zygotes were cultured in modification of CR1aa medium up to blastocyst and were fed serum 5 μl/50μl medium on day 6. Results of this experiment showed that the effect of concentration of glutathione (0, 0.25; 0.50; 0.75 and 1.00 mM) on fertilization medium to the percentage of cleavage rates were 69.35 + 29.40; 79.07 + 13.17; 67.88 + 10.65; 98.10 + 3.30 and 82.62 + 24.19% not significant different (P>0.05) among treatments A, B, C, D dan E respectively.The precentages of morula and blastocyst for treatment D were 50.45 + 42.64% and 31.28 + 24.28%; while 36.55 + 24.13% and 17.74 + 17.86% for treatment E respectively.   Key words: Glutathione, in vitro fertilization, oocytes, cleavage
The effects of proline, carnitine on the viability of sperm stored at 5oC (chilled semen) Polmer Situmorang; E Triwulanningsih; A Lubis; W Caroline; T Sugiarti
Jurnal Ilmu Ternak dan Veteriner Vol 6, No 1 (2001): MARCH 2001
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (147.85 KB) | DOI: 10.14334/jitv.v6i1.211

Abstract

An experiment was carried out to evaluate the addition of proline, carnitine in Tris-extender on the viability of bull sperm following storage at 5oC. Semen was collected by means of artificial vagina (AV), diluted in Tris-extender containing 5% V/V egg yolk (EY) and 4% V/V glycerol to get a final concentration of 50 million sperms/ml. Diluted semen cooled to 5oC for 45 minute and stored at those temperature for 1, 3, 10, and 13 days. In the first activity the addition of 15, 30 and 60 mM carnitine in Tris-extender while in the second activity the inclusion of 15, 30, and 60 mM proline on the viability of sperm was investigated. Addition of carnitine to Tris-extender signifinatly increase (P<0.05) the viability of sperm after storage for more than 3 days. At 3 days of storage, the mean %M and %L were 27.3, 38.8, 33.5, 53.0, 31.8, 47.0, and 30.5, 46.8 for control 15, 30, and 60 mM carnitine respectively. The similar results was obtained for 7 days of storage where the mean %M and %L for control (12.5 and 27.3) was significantly lower (P<0.05) than those 15, 30, and 60 mM carnitine (15.0, 33.5, 18.8, 36.5, 17.5, 36.3). The superiority of carnitine was maintained for 10 days of storage, where the mean %L were 23.5, 28.8, 31.5, and 30.3 for control; 15; 30; and 60 mM respectively. There was no any significant within concentration of carnitine tested (15 to 60 mM).The condition of apical ridge was not significantly affected by carnitine. In the second activity, inclusion of proline to Trisextender statistically (P<0.05) improved the viability of sperm after storage for 7 and 13 days. After 7 days of storage the mean %M and %L were 31.4, 36.4, 38.8, 40.4, 36.6, 42.7, and 34.8, 43.3 for control; 15, 30, and 60 mM proline respectively. The significant effects of proline was remain for 13 days of storage where the mean %M and % L were 24.6, 32.9, 28.6, 37.5, 29.1, 39.8, and 30.1, 37.3 for control; 15, 30, and 60 mM proline respectively. There was no significant difference within the concentration of proline. Condition of apical ridge was not significantly affected by proline.   Key words: Sperm, viability, carnitine, proline
Effect of exogenous glutathione in medium fertilization to improve blastocyst rates of bovine embryos E Triwulanningsih; P Situmorang; I-G Putu; T Sugiarti; A.M Lubis; D.A Kusumaningrum; W Caroline; R.G Sianturi
Jurnal Ilmu Ternak dan Veteriner Vol 7, No 2 (2002): JUNE 2002
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (190.218 KB) | DOI: 10.14334/jitv.v7i2.283

Abstract

Glutathione (C10H17N3O6S) is a tripeptide (γ-Glu-Cys-Gly) widespread in living organism. Glutathione (GSH) at the 5 mM concentration increased the motility and fertility of frozen-thawed sperm. Intracellulair glutathione improved the cleavage rate and embryo development to the blastocyst rate. Research on in vitro embryos production through the implementation of GSH during IVF (in vitro fertilization) on embryo development has been conducted at the Laboratorium Reproductive of Physiology, Research Institute for Animal Production. Ovaries of beef cattle as source of oocytes were collected from the slaughterhouse in a thermo flask with 350C PBS as medium and transported to the laboratory. The oocytes were fertilized in vitro with selected motile sperm using Percoll gradient (90 and 45%). Ten COCs (cumulus oocytes complexes) were transfered to 44 μl of fertilization medium (mTALP) was performed with either 0; 0.25; 0.50; 0.75 and 1.00 mM of glutathione as treatment A, B, C, D and E respectively, and inseminated with 2 μl of capacitated sperm and added 2 μl of heparin and 2 μl of PHE (consisting of 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine). Incubation between sperm and oocytes in the 5% CO2 incubator and RH 90% in fertilization media (TALP) for 20 hours. All zygotes were cultured in modification of CR1aa medium up to blastocyst and were fed serum 5 μl/50μl medium on day 6. Results of this experiment showed that the effect of concentration of glutathione (0, 0.25; 0.50; 0.75 and 1.00 mM) on fertilization medium to the percentage of cleavage rates were 69.35 + 29.40; 79.07 + 13.17; 67.88 + 10.65; 98.10 + 3.30 and 82.62 + 24.19% not significant different (P>0.05) among treatments A, B, C, D dan E respectively.The precentages of morula and blastocyst for treatment D were 50.45 + 42.64% and 31.28 + 24.28%; while 36.55 + 24.13% and 17.74 + 17.86% for treatment E respectively. Key words: Glutathione, in vitro fertilization, oocytes, cleavage