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All Journal Journal of the Indonesian Tropical Animal Agriculture
T. Maulana, T.
Research Centre for Biotechnology, Indonesian Institute of Sciences, Jl. Raya Bogor Km. 46 Cibinong 16911
Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences

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Journal : Journal of the Indonesian Tropical Animal Agriculture

 1 
STUDY ON CHANGES OF SPERM HEAD MORPHOMETRY AND DNA INTEGRITY OF FREEZE-DRIED BOVINE SPERMATOZOA Said, S.; Afiati, F.; Maulana, T.
Journal of the Indonesian Tropical Animal Agriculture Vol 40, No 3 (2015): September
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.40.3.145-152

Abstract

Changes of sperm heads morphometric and DNA integrity of freeze-dried bovine spermatozoa were investigated. Freeze-dried spermatozoa had stored in the refrigerator at 4°C for 2 years. Computer-assisted sperm analysis (CASA) was used in this study to identify sperm head morphometry, while for DNA integrity analysis using acridine orange staining. Samples were smeared on glass slides, fixed for 2 h in acetic alcohol and stained with acridine orange solution. After staining, each slide was examined at x400 magnification in a fluorescence microscope with axio vision (Zeiss Company, Germany). Proportion of fluorescence red and green emissions of the sperm head were examined and scored. These results indicated that sperm head had enlarged significantly (P<0.05) after freeze-drying process. However, freeze-dried sperm heads morphometry significantly (P<0.05) decrease after incubation for 3 and 6 hours. Changes of DNA integrity of freeze-dried spermatozoa significantly (P<0.05) decrease after incubation for 6 hours. In the present study concluded that (1) freeze-drying spermatozoa caused sperm head morphometric enlarged, whereas incubation time caused sperm heads decreased, (2) DNA integrity of freeze-dried sperm head is still intact during incubation 3 hours, and decreased DNA integrity occur in incubation for 6 hours.
Effect of addition an amino acid or its combination with EDTA on DNA integrity and morphometry sperm heads of freeze-dried bovine spermatozoa Said, S.; Maulana, T.; Setiorini, S.; Ibrahim, G.E.; Ramadhan, M.N.; Christopher, E.
Journal of the Indonesian Tropical Animal Agriculture Vol 45, No 3 (2020): September
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.45.3.189-196

Abstract

The objective of the current study was to investigate the effect of addition an amino acid or its combination with EDTA on DNA integrity and morphometry sperm heads of freeze-dried bovine spermatozoa. Spermatozoa were freeze-dried in medium with the addition of an amino acid glycine, cysteine, glutamine, or its combination with EDTA. Sperm head morphometry was identified at 400X magnification using Axio Vision(Zeiss Company, Germany), while for membrane plasma integrity (MPI) determined by calculation of the percentage of spermatozoa having intact plasma membrane by osmotic resistance test method and DNA integrity analysis using acridine orange staining. Sperm head had declined in size after the freeze-drying process, MPI of FD spermatozoa gradually increased significantly when FD solution was added with an amino acid solution (glycine, cysteine) and its combination with EDTA. DNA integrity of all freeze-dried spermatozoa treatments was remaining intact, no significantly different (P>0.01) among treatments. The present study concluded that the addition of an amino acid (glycine, cysteine) or its combination with EDTA could be reduced morphometric sperm heads and plasma membrane damage of freeze-dried bovine spermatozoa, however, DNA integrity of bovine sperm nucleus remaining intact after the freeze-drying process without addition both amino acids and EDTA. 
Sex sorting sperm of sumba ongole bulls by using snakehead fish (Channa striata) albumin extract Maulana, T.; Said, S.; Arifiantini, R. I.; Setiadi, M. A.
Journal of the Indonesian Tropical Animal Agriculture Vol 44, No 1 (2019): March
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.44.1.106-113

Abstract

The objective of this research was to investigate the potential of snakehead albumin extract (channalbumin) for sorting X and Y sperm of Sumba Ongole (SO) and its characteristic. Semen was collected from three SO bulls using artificial vagina and the freeze dried channalbumin was extracted from snakehead fish. Channalbumin column was made with different concentration ratio of top and bottom fraction: 2%:4%; 3%:5%; 4%:6% respectively and BSA 5%:10% as control. Semen was put in top fraction then incubated for 30 min at room temperature then each fraction was centrifuged at 1800 rpm for 10 minutes. The pellet was evaluated for motility, abnormality, viability, membrane integrity and head sperm morphometric. The results showed that the channalbumin capable to maintain sperm motility in the top fraction better than the bottom fraction. Sperm viability and membrane integrity in control group were significantly higher (P<0.05) than all channalbumin treatment. BSA 5%:10% has highest proportion of X and Y sperm (69%:76.77%) compared with 2%:4% (42.33%:79.13%), 3%:5% (55.97%:75.73%) and 4%:6% of channalbumin (62.77%:68%). It’s concluded that channalbumin 4%: 6% was effective for separation of XY sperm with higher proportion.
Co-Authors Christopher, E. F. Afiati, F. Ibrahim, G.E. M. A. Setiadi, M. A. R. I. Arifiantini Ramadhan, M.N. S. Said Setiorini, S.