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All Journal Jurnal Akuakultur Indonesia Jurnal Natur Indonesia Journal of Aquaculture Management and Technology
Ayi Santika
Balai Besar Pengembangan Budidaya Air Tawar (BBPBAT) Sukabumi, Jl. Selabintana No. 37 kotak pos 67 Kota Sukabumi 43114
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Decision Sciences, Operations Research & Management Other

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Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio) Nuryati, Sri; Alimuddin, Alimuddin; Sukenda, Sukenda; Soejoedono, Retno Damayanti; Santika, Ayi; Pasaribu, Fachriyan Hasmi; Sumantadinata, Komar
Jurnal Natur Indonesia Vol 13, No 1 (2010)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (120.942 KB) | DOI: 10.31258/jnat.13.1.47-52

Abstract

Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.
Aplikasi Vaksin Dna Koi Herpes Virus (Khv) Melalui Metode Perendaman Dengan Dosis Yang Berbeda Dalam Upaya Pencegahan Penyakit Pada Ikan Mas (Cyprinus carpio) Chairunnisa, Sekar Ayu; Prayitno, Slamet Budi; -, Sarjito; Santika, Ayi; Nuryati, Sri
Journal of Aquaculture Management and Technology Vol 2, No 2 (2013) : Journal of Aquaculture Management and Technology
Publisher : Journal of Aquaculture Management and Technology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1457.163 KB)

Abstract

The purpose of this research was to determine fish clinical signs of KHV-infected and determine effectiveness of KHV DNA vaccine with immersion method to prevent koi herpes virus (KHV) infection. This research was conducted in National Center for Freshwater Aquaculture Development (BBPBAT) Sukabumi, from October 2012 to Februari 2013. The fish samples used were carp seed free KHV with size 3 – 5 cm as many as 450 animals. Five (5) treatments and 3 replications were administered in this research. The treatment were, without vaccine, vaccine at dose of 104, 105, 106, and 107 CFU/ml. Vaccination technique used was vaccination through immersion method. One of the vaccine that has been developed was KHV DNA vaccine. Eksperimental fishes were maintain for 30 days past vaccination, and then challenged with KHV and observed for 21 days. The variables measured were clinical signs and survival rate. The results of this research indicated that non vaccinated fish (A), vaccinated fish at 104(B) and 105(C). demonstered clinical signs of KHV-infected fish, while vaccinated fish at 106(D) and 107(D) visibly normal. This results indicated that vaccination using KHV DNA vaccines at 106 and 107 CFU/ml could prevent eksperimental fish from KHV-infection. PCR results showed that fish with treatments A, B and C positive infected KHV, while treatment D and E negative KHV. Survival rate (SR) at treatment A (15,56%), B (50,00%), C (53,33%), D (78,89%) and E (83,33%). For prevention of KHV, dose vaccine DNA KHV 106 and 107 CFU/ml could be used. This results supported by survival rate from treatment D (78,89%) and E (83,33%) higher than treatment A, B and C.
Gambaran Parameter Hematologis Pada Ikan Nila (Oreochromis niloticus) Yang Diberi Vaksin DNA Streptococcus iniae Dengan Dosis Yang Berbeda Utami, Devitha Tri; Prayitno, Slamet Budi; Hastuti, Sri; Santika, Ayi
Journal of Aquaculture Management and Technology Vol 2, No 4 (2013) : Journal of Aquaculture Management and Technology
Publisher : Journal of Aquaculture Management and Technology

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Abstract

Tilapia (O. niloticus) is one of freshwater species that has rapid growth and easily been cultivated. One of the constraints in cultivating Tilapia is Streptococciasis disease caused by Streptococcus iniae. The purpose of this research was to determine the haematological response of Tilapia that was given by DNA vaccine of S. iniae and challenged with S. iniae, and to determine the most effective dose of DNA vaccine based on haematological performances in the prevention of bacteria S. iniae. The fish samples used were Tilapia with length of 7,79 cm ± 0.48 as many as 150 animals or 10 animals per treatment. Five (5) treatments and three (3) replications were administered in this research, there were A (0 ŋg/µL), B (10ŋg/µL), C (20ŋg/µL), D (30ŋg/µL), dan E (40ŋg/µL) of DNA vaccines the injected intramuscularly. Fish were maintained for 30 days, then challenged with S. iniae for 14 days. Measurement of haematological performances include haematocrit, total leukocyte, differential leukocytes, and phagocytosis index carried out every 7 days. The observation of clinical signs of fish after challenge by S. iniae obtained that fish in A and B treatment showed clinical signs more quickly than fish in C, D, and E treatment. The results showed that DNA vaccines S. iniae in E treatment significantly different on haematocrit levels (25,67%), total leukocytes (9,46x104 cells/mm3), lymphocytes (80,33%), monocytes (5,67%), neutrophils (14%), phagocytosis index (22,33%), and survival rate (70%). It can be conclude that dosage of DNA vaccine 40ng/μl is the most effective dosage which is immunogenic and protective in raising Tilapia immunity against bacterial S. iniae based on haematological aspects.
Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion Nuswantoro, Soko; Alimuddin, .; Yuhana, Munti; Santika, Ayi; Nuryati, Sri; Zainun, Zakki; Mawardi, Mira
Jurnal Akuakultur Indonesia Vol. 11 No. 1 (2012): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (991.677 KB) | DOI: 10.19027/jai.11.76-85

Abstract

Koi herpesvirus (KHV) is a herpesvirus that particularly infects and causes mass mortality to koi and common carp. Therefore, the protection of common carp from KHV infection is urgently needed. In this study, we developed an application of DNA vaccine encoding KHV glycoprotein-25 by immersion method to increase survival of common carp against KHV infection. A total of 400 common carp juveniles at 30-day-old were immersed in 1-L water containing 1.3×108CFU/mL of the killed Escherichia coli cells carrying DNA vaccine. Three frequencies and three duration of fish immersion were tested, namely: 1×30 minutes, 1×60 minutes, 1× 90 minutes, 2×90 minutes and 3×90 minutes by interval of 24 hours. Reversetranscription polymerase chain reaction analysis showed that DNA vaccine was successfully expressed in the vaccinated fish. Fish at twenty eight days post vaccination were challenged by injecting 10-4 mL of KHV per fish. The result showed that vaccination by 1×30 minutes immersion allowed 61% of fish survived, and this was significantly higher (p<0.05) compared to control (without vaccination), but it was similar among vaccination treatments (p>0.05). The relative percent survival of vaccinated fish were also similar among treatments (p>0.05). DNA vaccination has increased fish survival about two fold higher compared to unvaccinated fish control (26.67%). Thus, DNA vaccination was effectively delivered by immersion for 1×30 minutes, and this technique can be useful to level up the resistance of common carp juveniles against KHV infection. Keywords: DNA vaccine, KHV, glycoprotein, immersion, common carp
Co-Authors Alimuddin Devitha Tri Utami Fachriyan Hasmi Pasaribu Komar Sumantadinata Laksono Trisnantoro Mira Mawardi MUNTI YUHANA Retno Damayanti Soejoedono Sarjito - Sekar Ayu Chairunnisa Slamet Budi Prayitno Soko Nuswantoro Sri Nuryati Sukenda . Zakki Zainun