N. W. K. Karja
Division of Reproduction and Obstetric, Department of Veterinary Clinic, Reproduction and Pathology, Faculty of Veterinary Medicine, Bogor Agricultural University

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Journal : Tropical Animal Science Journal

Cryopreservation of Swamp Buffalo Semen in Skim Milk Yolk-based Diluent with Two Different Cryoprotectants M. T. Herbowo; R. I. Arifiantini; N. W. K. Karja; R. G. Sianturi
Tropical Animal Science Journal Vol. 42 No. 1 (2019): Tropical Animal Science Journal
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (284.378 KB) | DOI: 10.5398/tasj.2019.42.1.13

Abstract

The successful of artificial insemination (AI) in buffalo is still low. The quality of frozen semen determines the successful of AI program. Glycerol is widely used for cryopreservation of buffalo semen with unsatisfactory results due to its toxicity and antifertility properties. This study aimed to investigate the effect of glycerol and dimethylformamide (DMF) concentrations in skim milk egg yolk (SMEY)–based diluent on the quality of frozen-thawed swamp buffalo bull semen. Fresh semen was divided into eight aliquots. Each aliquot was diluted in SMEY containing 4%, 5%, 6%, and 7% glycerol and 4%, 5%, 6%, and 7% DMF. Diluted semen was packed into mini straw (0.25 mL), equilibrated at 4 °C for 4 hours, frozen in liquid nitrogen vapor for 10 minutes, and stored at liquid nitrogen container. The straws were thawed after 24 hours at 37 °C for 30 seconds for evaluation. The results showed that the post thawed sperm motility and recovery rate in SMEY extender containing 7% glycerol and 5% DMF were higher than the other glycerol or DMF concentrations. Post thawed sperm viability and plasma membrane integrity in SMEY containing 4%-6% DMF did not differ significantly, however both values were higher than those in 7% DMF. Post thawed sperm viability and plasma membrane integrity did not differ between SMEY containing 6% and 7% glycerol. However, both values were higher than those in 4% and 5% glycerol. It is concluded that 7% glycerol or 5% DMF is considered to be the most suitable cryoprotective agent for swamp buffalo semen cryopreservation.The successful of artificial insemination (AI) in buffalo is still low. The quality of frozen semen determines the successful of AI program. Glycerol is widely used for cryopreservation of buffalo semen with unsatisfactory results due to its toxicity and antifertility properties. This study aimed to investigate the effect of glycerol and dimethylformamide (DMF) concentrations in skim milk egg yolk (SMEY)–based diluent on the quality of frozen-thawed swamp buffalo bull semen. Fresh semen was divided into eight aliquots. Each aliquot was diluted in SMEY containing 4%, 5%, 6%, and 7% glycerol and 4%, 5%, 6%, and 7% DMF. Diluted semen was packed into mini straw (0.25 mL), equilibrated at 4 °C for 4 hours, frozen in liquid nitrogen vapor for 10 minutes, and stored at liquid nitrogen container. The straws were thawed after 24 hours at 37 °C for 30 seconds for evaluation. The results showed that the post thawed sperm motility and recovery rate in SMEY extender containing 7% glycerol and 5% DMF were higher than the other glycerol or DMF concentrations. Post thawed sperm viability and plasma membrane integrity in SMEY containing 4%-6% DMF did not differ significantly, however both values were higher than those in 7% DMF. Post thawed sperm viability and plasma membrane integrity did not differ between SMEY containing 6% and 7% glycerol. However, both values were higher than those in 4% and 5% glycerol. It is concluded that 7% glycerol or 5% DMF is considered to be the most suitable cryoprotective agent for swamp buffalo semen cryopreservation.
Efficacy of Leptin Supplementation on Nuclear Maturation and Fertilization Rate of Sheep Oocytes In Vitro A. Riyuska; N. W. K. Karja; A. Setiadi
Tropical Animal Science Journal Vol. 42 No. 1 (2019): Tropical Animal Science Journal
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (459.684 KB) | DOI: 10.5398/tasj.2019.42.1.1

Abstract

The objective of this study was to determine the efficacy of leptin supplementation into maturation medium on nuclear maturation and fertilization rate of sheep oocytes. The maturation process was conducted using a tissue culture medium (TCM) 199 with four supplementation treatments of leptin namely 0 (control), 10, 50, and 100 ng/mL. Fertilization was conducted in the oocytes supplemented with 10 ng/mL and control using 5×106 mL-1 spermatozoa. At the end of maturation and fertilization processes, the oocytes were stained with 2% aceto orcein to determine nuclear maturation rate and pronuclear development. The results showed that the percentage of oocytes reaching metaphase II (MII) stage significantly increased in the oocytes supplemented with leptin at a dose of 10 ng/mL (P<0.05) compared to those supplemented at doses of 50, 100, and 0 ng/mL (93.7±5.9% vs 78.8±4.4%; 72.0±2.6%; 82.1±9.9%). However, fertilization rate of the oocytes supplemented with leptin at a dose of 10 ng/mL and control were similar (72.1±5.5% vs 79.2±7.0%). The data indicated that leptin could improve maturation rate in lower concentration. However, the improved maturation rate of oocytes with leptin supplementation at a dose of 10 ng/mL could not improve the fertilization rate of the oocytes. In conclusion, the supplementation of leptin at a dose of 10 ng/mL could increase the number of oocytes that reached MII stage, but could not increase the fertilization rate.