Judul: Optimization of Electroporation with Green Fluorescent Protein Gene for Testicular Cell Transplant in Nile Tilapia
Abstrak: Transplantation technology can be applied to generate surrogate fish broodstock. Germinal cells as donor used in transplantation are labeled to distinguish with endogenous cells. Donor cells are generally derived from transgenic fish carrying a marker or cells labeled by PKH-26. This study was performed to obtain an alternative method of labeling cells by using electroporation. The method in this study includes the preparation of the recipient, the donor cell preparation and cell transplantation. Recipient was obtained by mating tilapia males and females with a ratio of 1:1. Testicular cells were taken from 4-month-old Nile tilapia. Electroporation was performed with testicular cell density of 104 cells.μL-1, pJfKer-GFP concentration of 50 ng.μL-1, and a pulse length of 20 ms at 0, 100, 200, and 300 volts. In amount of 5x103 cells at 0.5 μL electroporated testicular cells were then injected into the intraperitoneal cavity of 3-day-old Nile tilapia larva. The parameters measured were cell survival, the percentage of GFP fluorescent cells, and cell colonization in recipient fish. The results showed that survival of the electroporated cells with 100 and 200 volts was similar (p>0.05), and higher than that of 300 volt treatment (p<0.05). It shows that the voltage is very influential on cell survival. The increase in voltage will result in the increasing mortality in the cell. Number of fluorescent cells was not significantly different among treatments. While the percentage of fluorescent cells did not differ on the treatment of 100, 200, and 300 volts (p> 0.05) except for the control. In the control treatment luminescence GFP was not visible, it is proved that the GFP gene was not able to pass through the cell membrane in the absence of an electric voltage. Percentage colonization of transplanted cells in the fish after 50 days of rearing gave the highest value at 200 volts (66.67%). As conclusion, electroporation within 100-200 volt was a suitable tool to label testicular cells for transplantation.
Keyword: electroporation, GFP, label, Nile tilapia, transplantation

Judul: Transplantasi sel testikular ikan gurame pada ikan nila
Abstrak: One of the constraints in the cultivation of giant gouramy is the lack of fry availability, necessitating the development of fry production engineering technology to address these issues. Engineering technology of fry production through the surrogate broodstock generated by testicular cells transplantation technique has been developed in salmonid. Testicular cells transplantation technology requires a suitable recipient that can accept and support the development of spermatogonia donor cell. This study examined the potential of Nile tilapia as a recipient for donor giant gouramy cells by observing its colonization in the gonads of Nile tilapia. Fish donor of 598,82 grams body weight with Gonado Somatic Index (GSI) of 0,013% and spermatogonial cells percentage of 28,22% was used. Testicular cells in amount of 20.000 cells/0,5 μl PBS were injected into peritoneal cavity of 4-day-old tilapia larvae. The success of transplantation and donor cell colonization were analyzed using PCR method with DNA template that have been extracted from tilapia larvae of 1 day post injection and the gonad of 2 months old fish. PCR was performed using a set of specific primer for the giant gouramy growth hormone gene and tilapia β-actin as an internal control of DNA loading. PCR results showed that the transplanted tilapia has a DNA band in the same size with giant gouramy, and not in untransplanted control. The success rate of transplantation in larvae was 100% (5/5) and the percentage of individual recipients containing donor cells in their gonad was 60% (3/5). This indicates that donor cells from giant gouramy was successfully injected and can be colonized in the gonads of Nile tilapia, so the Nile tilapia potentially be used as recipient for giant gouramy testicular cells transplantation. Further analysis needs to be conducted to observe the ability of spermatogonial cells develop into sperm and eggs in the gonad of tilapia recipient.
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Judul: Analisis keamanan konstruksi fungsi hash berbasis sandi blok kunci simetrik (Kajian Teoritik)
Abstrak: Hash function is used to secure the integrity and authenticity of a massege, so that the message is save. The message which will be secured, will be attached to hash/message digest/digital signature value fro, the message. A secure hash function is the one which has pre-image resistance, 2nd pre-image resistance and collision resistance. Hash function has been classified into hash keyed function (Message Authentication Codes/MAC) and unkeyed (Modification Detection Codes/MDC)
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