Ike Trismawanti
Balai Riset Perikanan Budidaya Air Payau dan Penyuluhan Perikanan, Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

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EFFECT OF VARIOUS CRYOPROTECTANTS ON PRESERVATION OF BLACK TIGER Penaeus monodon SHRIMP SPERMATOZOA Rosmiati Rosmiati; Ike Trismawanti; Samuel Lante
Indonesian Aquaculture Journal Vol 15, No 2 (2020): (December, 2020)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/iaj.15.2.2020.77-83

Abstract

The development of cryopreservation technique on tiger shrimp Penaeus monodon broodstock spermatophore has been carried out to support the artificial insemination. This study aims to determine the effect of three cyoprotectants (methanol, dimetylsulphoxide (DMSO), and glycerol) for long term storage of tiger shrimp Penaeus monodon spermatozoa. Spermatophores were collected from the wild broodstocks through electrical shock. Spermatozoa were obtained by homogenizing the spermatophores using a Radnoti micro homogenizer in Ca-free saline solution containing one of three cryoprotectans (methanol, dimetylsulphoxide, and glycerol) separately at the concentration of 5%. One mL of each cryoprotectant containing spermatozoa with the density of 1.02 x 106 cell/mL was transferred into a cryovial and cryopreserved at room temperature, -20°C and -196°C for 5, 10, and 30 days. The apparent sperm viability (ASV) of cryopreserved spermatozoa was monitored after treated. Thawing of cryopreserved spermatozoa was carried out in a 30°C wáter bath for two minutes. The result showed that the best apparent sperm viability was obtained at the using of glycerol at -196°C in liquid nitrogen, even after the thirty days of cryopreservation time period with the ASV of 0.82 x 106 cells/mL (80.39%). Meanwhile two other cryoprotectans displayed the ASV of 0.54 x 106 cells/mL (56.86%), and 0.23 x 106 cells/mL (22.55%). for DMSO and methanol, respectively. In turn, the control showed the lowest ASV with the ASV of 0.01 x 106 cells/mL (1.27%). The ASV showed by this glycerol exhibited asignificant difference (P<0.05) to that of methanol, DMSO, and control.
RESPONS IMUN UDANG WINDU Penaeus monodon TERHADAP VAKSIN dsRNA VP-24 PADA DOSIS BERBEDA Sri Redjeki Hesti Mulyaningrum; Andi Parenrengi; Bunga Rante Tampangallo; Ike Trismawanti
Jurnal Riset Akuakultur Vol 13, No 1 (2018): (Maret 2018)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (463.845 KB) | DOI: 10.15578/jra.13.1.2018.77-84

Abstract

Peningkatan produksi udang windu Penaeus monodon terus diupayakan, salah satunya dengan peningkatan respons imun udang terhadap infeksi penyakit WSSV. Penelitian ini bertujuan untuk mengetahui respons imun udang terhadap pemberian vaksin dsRNA VP-24 pada berbagai dosis. Konstruksi vaksin dsRNA VP-24 dilakukan menggunakan Megascript kit dengan DNA genom VP-24 sebagai template. Vaksinasi dilakukan dengan metode injeksi pada udang windu yang berukuran rata-rata 15,88 ± 3,50 g; dosis vaksin yang diujikan adalah 0,02 µg; 0,2 µg; 2 µg; dan sebagai kontrol adalah udang yang tidak diberi vaksin. Penelitian terdiri atas empat perlakuan dosis vaksin dengan masing-masing dua ulangan dan dipelihara selama lima hari. Uji tantang dilakukan selama enam hari dengan menginjeksi virus WSSV dalam saline solution (1:3 v/v). Pengamatan terhadap sintasan udang windu dilakukan setiap hari, sedangkan penghitungan total hemocyte (THC) dan ProPO diamati pada hari I, III, dan VI setelah diinfeksi WSSV. Pada akhir pengujian dilakukan pengambilan jaringan hepatopankreas untuk analisis histopatologi. Analisis data dilakukan secara statistik dengan analisis ragam (ANOVA). Hasil yang diperoleh memperlihatkan bahwa injeksi vaksin dsRNA VP-24 dengan dosis 0,2 µg memiliki pengaruh yang signifikan terhadap sintasan dan respons imun udang windu (P<0,05). Vaksin dsRNA VP-24 dengan dosis 0,2 µg mampu memberikan sintasan udang windu P. monodon sebesar 65% dan meningkatkan respons imun udang dengan THC (1.550 x 10t sel/mL) dan ProPO (0,042 Abs).One of the efforts to increase the production of tiger shrimp Penaeus monodon is increasing the immune response against WSSV disease. This study aims to evaluate shrimp immune response to dsRNA VP-24 vaccination at various doses. The construction of dsRNA VP-24 vaccine was performed using Megascript kit with the VP-24 DNA genome as a template. The vaccination was done by injection method on shrimp sized 15.88 ± 3.50 g. The tested vaccine doses (treatments) were 0.02 ¼g; 0.2 ¼g; 2 ¼g; and unvaccinated shrimp as the control. The study consisted of four treatments of vaccine doses with two replicates for each treatment. The challenge test was performed by injecting the WSSV virus in saline solution (1:3 v/v). The observation on shrimp survival rate was done daily, while the total hemocyte count (THC) and ProPO observation were performed on the 1st day, 3rd day, and 6th day after WSSV infection. At the end of the experiment, samplings of hepatopancreas for analysis were performed. Data were statistically analyzed using ANOVA. The present study indicated that the injection of 0.2 ¼ g dsRNA VP-24 vaccine had a significant effect on the survival rate and immune response of shrimp (P<0.05). The dose of 0.2 ¼g dsRNA VP-24 had resulted in 65% of survival rate and increased immune response of P. monodon with THC (1,550 x 10t cell/mL) and ProPO (0.042 Abs).