Hematology of common carp following DNA vaccination  and koi herpesvirus challenge test

The study was aimed to determine the effectiveness of DNA vaccine doses on hematological aspect which represent immune response and its influence on common carp survival rate. DNA vaccines encoding the viral glycoprotein of koi herpesvirus (KHV) have been proved to highly protect the fish under laboratory condition. A dose of 12.5 µg/100 µl vaccine had resulted in a survival rate of 96.67 % during 30 days after challenge test with a lethal dose of KHV. Fish vaccinated using lower doses, i.e. 2.5 and 7.5 µg/100µl showed 100% mortality after 15 and 19 days challenge test respectively, whereas non vaccinated fish as a control showed 100% mortality after 17 days challenge test. Total leucocytes of the vaccinated fish were higher than control until 42 days post vaccination, but declined afterward. Phagocytic index of the vaccinated fish using 12.5 µg/100 µl was declined after 49 days post vaccination or 7 days post challenge test. Key words: DNA vaccine, Koi herpesvirus (KHV), leucocyte, phagocytic index, Cyprinus carpio ABSTRAK Penelitian ini bertujuan untuk menguji pengaruh vaksinasi menggunakan vaksin DNA dengan dosis berbeda terhadap gambaran darah ikan sebagai respresentasi tanggap kebal ikan mas serta pengaruhnya terhadap tingkat kelangsungan hidup ikan mas. Vaksin DNA penyandi glikoprotein koi herpesvirus (KHV) dapat memberikan proteksi yang tinggi pada percobaan skala laboratorium. Vaksinasi dengan dosis 12,5 µg/100µl dapat mempertahankan kelangsungan hidup sebesar 96,67% selama satu bulan setelah uji tantang dengan virus KHV menggunakan dosis letal. Ikan yang divaksin dengan dosis yang lebih rendah yaitu 2,5 dan 7,5 µg/100µl mengalami kematian total berturut-turut setelah 15 dan 19 hari uji tantang, sedangkan ikan kontrol yang tidak divaksin mengalami kematian total setelah 17 hari uji tantang. Jumlah leukosit total ikan yang divaksinasi lebih tinggi dibanding dengan kontrol sampai hari ke-42, setelah itu mengalami penurunan. Indeks fagositosis ikan yang divaksin dengan dosis 12,5 µg/100µl mengalami penurunan setelah hari ke-49 atau 7 hari setelah uji tantang. Kata kunci: Vaksin DNA, Koi herpesvirus (KHV), leukosit, indeks fagositosis, Cyprinus carpio

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Efficacy of GP-11 KHV DNA Vaccine in Common Carp (Cyprinus carpio) through Feed by Different Frequency of Administration

Omni-Akuatika ◽

10.20884/1.oa.2020.16.1.768 ◽

2020 ◽

Vol 16 (1) ◽

pp. 1

Author(s):

Ahmad Beni Rouf ◽

Sri Nuryati ◽

Sukenda Sukenda ◽

Alimuddin Alimuddin

Keyword(s):

Dna Vaccine ◽

Common Carp ◽

White Blood Cells ◽

Challenge Test ◽

Positive Control ◽

Negative Control ◽

Lysozyme Activity ◽

Post Vaccination ◽

Koi Herpesvirus ◽

Vaccine Treatment

GP-11 KHV DNA vaccine is a vaccine that can be used to induce immunity against the KHV virus (Koi herpesvirus). Vaccination through feed is an alternative way of administering vaccines. The study aimed to examine the effect of giving KHV GP-11 DNA vaccine through feed with different frequencies to KHV infection. The frequency of vaccine administration is GP-11 vaccination once a week; GP-11(1x), GP-11 vaccination twice a week; GP-11(2x), GP-11 vaccination three times a week; GP-11(3x), GP-25 vaccinations three times a week; GP-25(3x), negative control (without KHV test) and positive control (KHV tested). The fish were kept for 28 days after vaccination and then continued with the KHV challenge test for 28 days. The weight of carp ranges from 13.82±2.37 g maintained with a density of 15 fish/aquarium. The results showed that vaccine treatment was able to induce an immune response as indicated by the number of white blood cells, lysozyme activity and post-vaccination antibody titer showed a significant effect compared to controls. Likewise, after the challenge test, supported by IFNγ and IgM gene expression parameters after the challenge test showed the highest value of vaccine treatment rather than control. The efficacy of vaccine was showed by RPS value (%) in each vaccine treatment obtained GP-11(1x) value of 44.7±3.7a, GP-11(2x) of 78.9±18.2b, GP-11(3x) 85.6±12.6b and GP-25(3x) 79.5±18.1b. It was concluded that administering the GP-11 vaccine frequency 2 times a week provides protection as strong as giving a vaccine frequency 3 times a week.Keywords: common carp, DNA vaccine, frequency of administration, koi herpesvirus

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The Effectiveness of Onion Extract Allium sativum to Prevent Koi Herpesvirus (KHV) Infection on Common Carp Cyprinus carpio

Jurnal Akuakultur Indonesia ◽

10.19027/jai.7.139-150 ◽

2008 ◽

Vol 7 (2) ◽

pp. 139

Author(s):

Sri Nuryati ◽

P. Giri ◽

Y. Hadiroseyani

Keyword(s):

Immune System ◽

Survival Rate ◽

Common Carp ◽

Cyprinus Carpio ◽

Allium Sativum ◽

Garlic Extract ◽

Herpesvirus Infection ◽

Onion Extract ◽

Koi Herpesvirus ◽

Herpès Virus

Common carp is one of consumption fish that has delicious meat, high pritein level, and easy in farming. The serious problem in common carp farming is koi herpesvirus infection. Onion extract potency to improve immune system was estimated to prevent disease infection. The testing of the garlic extract through food could be used as efforts to increase endurance of common carp fish Cyprinus carpio to koi herpesvirus infection that was considered from blood parameter. Fish that was used was measuring 9-11 cm with the treatment of food containing 30, 50, and 70 gr/100 ml onion extract. Fish was acclimated for seven days in 60×30×30 cm3 aquarium before used. Garlic extract diet in food gave increasing of fish immune system that was infected by koi herpesvirus. The increased of leucocytes of blood fish with onion extract diet was faster than possitive control. The dose of B treatment (50 gr/100 ml) was the best dose gave short incubation periode comparing other treatment. Survival rate (SR) of this B treatment was highest, i.e. 91.7%, while survival rate of negative control was 50%. Key word: common carp, Cyprinus carpio, onion, Allium sativum, koi herpesvirus ABSTRAK Salah satu jenis ikan konsumsi air tawar yang banyak digemari oleh masyarakat adalah ikan mas Cyprinus carpio karena rasa dagingnya gurih, memiliki kadar protein tinggi dan cukup mudah dalam pemeliharaannya. Permasalahan yang muncul saat ini adalah wabah Koi Herpes Virus (KHV). Potensi ekstrak bawang putih sebagai anti mikroba spektrum luas, diduga dapat mengobati dan mencegah penyakit ikan. Pengujian bawang putih secara in vivo melalui pakan dapat digunakan sebagai upaya untuk meningkatkan ketahanan tubuh ikan mas Cyprinus carpio terhadap infeksi penyakit KHV yang ditinjau dari gambaran darahnya. Ikan uji yang digunakan adalah ikan mas berukuran 9-11 cm dengan perlakuan pakan yang mengandung bawang putih sebanyak 30, 50, dan 70 gr/100 ml. Sebelum dilakukan penelitian ikan diadaptasikan selama 7 hari pada akuarium berukuran 60×30×30 cm3. Hasil penelitian menunjukan bahwa ekstrak bawang putih efektif untuk meningkatkan ketahanan tubuh ikan mas Cyprinus carpio yang diinfeksi oleh Koi Herpes Virus (KHV). Pengamatan gambaran darah ikan yang terinfeksi KHV setelah pemberian ekstrak bawang putih selama 30 hari dapat meningkatkan jumlah leukosit lebih cepat dibandingkan dengan perlakuan kontrol positif. Peningkatan jumlah limfosit dalam darah mampu meningkatkan pertahanan tubuh. Peningkatan jumlah leukosit ditunjukkan dengan meningkatnya jumlah limfosit dan penurunan jumlah monosit. Perlakuan B (50 gr/100 ml) merupakan dosis yang paling baik karena masa inkubasi KHV terlewati lebih cepat dibanding perlakuan lain yang ditandai dengan penurunan jumlah leukositnya. Hal ini didukung oleh gejala klinis yang ringan dibandingkan kontrol positif dan tingkat kelangsungan hidup yang tinggi sebesar 91,7%. Sedangkan pada kontrol negatif kelangsungan hidupnya sebesar 50%. Kata kunci: ikan mas, Cyprinus carpio, bawang putih, Allium sativum, KHV dan Herpes

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Intra-muscular and oral vaccination using a Koi Herpesvirus ORF25 DNA vaccine does not confer protection in common carp (Cyprinus carpio L.)

Fish & Shellfish Immunology ◽

10.1016/j.fsi.2018.03.037 ◽

2019 ◽

Vol 85 ◽

pp. 90-98 ◽

Cited By ~ 9

Author(s):

Carmen W.E. Embregts ◽

Roni Tadmor-Levi ◽

Tomáš Veselý ◽

Dagmar Pokorová ◽

Lior David ◽

...

Keyword(s):

Dna Vaccine ◽

Common Carp ◽

Cyprinus Carpio ◽

Oral Vaccination ◽

Koi Herpesvirus ◽

Common Carp Cyprinus Carpio ◽

Carp Cyprinus Carpio

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Potential transmission test of GP25 vaccine in normal flora bacteria of common carp culture media

Jurnal Akuakultur Indonesia ◽

10.19027/jai.14.90-97 ◽

2015 ◽

Vol 14 (1) ◽

pp. 90

Author(s):

Sri Nuryati ◽

, Alimuddin ◽

Ayu Dhita Juliadiningtyas

Keyword(s):

Dna Vaccine ◽

Common Carp ◽

Cyprinus Carpio ◽

Dna Vaccines ◽

Culture Media ◽

Pond Water ◽

Vaccine Safety ◽

Normal Flora ◽

Koi Herpesvirus ◽

Transmission Test

ABSTRACT Koi herpesvirus (KHV) is a virus that infects common carp and koi Cyprinus carpio. KHV attacks most stadia of common carp and koi and causes mortality up to 80–95% of the population. One way to prevent the spread of KHV is by applying DNA vaccine. This research was conducted to test potential transmission of DNA vaccine encoding glycoprotein 25 (vaccine GP25) toward normal flora bacteria of media cultivation of common carp. Bacteria was isolated from pond water of common carp and tested for their sensitivity to ampicillin. Research was performed by adding vaccine GP25 to ampicillin-sensitive bacteria at a dose of 12.5 µg/100 µL and incubated at 28 °C for 30, 60, 180, and 300 minutes then plated on media containing ampicillin. The grown bacteria cells were tested for the existence of plasmid bearing gen GP25 through bacteria colony cracking. The results of this research showed that there was no bacteria contained plasmid bearing gen GP25. Keywords: DNA vaccines, common carp, vaccine safety ABSTRAK Koi herpesvirus (KHV) merupakan virus yang menginfeksi ikan mas dan koi Cyprinus carpio. KHV menyerang hampir semua stadia ikan mas dan koi dan menyebabkan kematian hingga 80–95% dari populasi. Salah satu cara penanggulangan penyebaran KHV adalah dengan penggunaan vaksin DNA. Penelitian ini dilakukan untuk menguji potensi transmisi vaksin DNA glikoprotein 25 (vaksin GP25) pada bakteri flora normal pada media budidaya ikan mas. Bakteri diisolasi dari kolam budidaya ikan mas dan diuji sensitivitasnya terhadap antibiotik ampisilin. Penelitian dilakukan dengan menambahkan vaksin GP25 ke larutan bakteri sensitif ampisilin dengan dosis 12,5 µg/100 µL dan diinkubasi pada suhu 28 °C selama 30, 60, 180, dan 300 menit kemudian disebar pada media mengandung antibiotik ampisilin. Sel bakteri yang tumbuh diuji untuk mendeteksi keberadaan plasmid pembawa gen GP25 dengan menggunakan metode seleksi koloni bakteri (colony cracking). Hasil penelitian ini menunjukkan bahwa tidak ada bakteri yang mengandung plasmid pembawa gen GP25. Kata kunci: keamanan vaksin, ikan mas, vaksin DNA

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Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion

Jurnal Akuakultur Indonesia ◽

10.19027/jai.11.76-85 ◽

2013 ◽

Vol 11 (1) ◽

pp. 76 ◽

Cited By ~ 2

Author(s):

Soko Nuswantoro ◽

. Alimuddin ◽

Munti Yuhana ◽

Ayi Santika ◽

Sri Nuryati ◽

...

Keyword(s):

Dna Vaccine ◽

Common Carp ◽

Polymerase Chain Reaction Analysis ◽

Dna Vaccination ◽

Chain Reaction ◽

Immersion Method ◽

Post Vaccination ◽

Koi Herpesvirus ◽

Polymerase Chain ◽

Fish Survival

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

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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)

Jurnal Natur Indonesia ◽

10.31258/jnat.13.1.47-52 ◽

2012 ◽

Vol 13 (1) ◽

pp. 47 ◽

Cited By ~ 1

Author(s):

Sri Nuryati ◽

Alimuddin Alimuddin ◽

Sukenda Sukenda ◽

Retno Damayanti Soejoedono ◽

Ayi Santika ◽

...

Keyword(s):

Survival Rate ◽

Dna Vaccine ◽

Common Carp ◽

Cyprinus Carpio ◽

Vaccine Development ◽

Fish Muscle ◽

Negative Control ◽

Control Fish ◽

Pcr Analysis ◽

Fish Group

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.

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INDUKSI KEKEBALAN SPESIFIK PADA IKAN MAS, Cyprinus carpio Linn. TERHADAP INFEKSI KOI HERPESVIRUS (KHV) MELALUI TEKNIK KOHABITASI TERKONTROL

Jurnal Riset Akuakultur ◽

10.15578/jra.5.2.2010.257-276 ◽

2016 ◽

Vol 5 (2) ◽

pp. 257

Author(s):

Taukhid Taukhid ◽

Angela Mariana Lusiastuti ◽

Wulan Andiyani ◽

Rosidah Rosidah ◽

Sriati Sriati

Keyword(s):

Survival Rate ◽

Common Carp ◽

Cyprinus Carpio ◽

Clinical Signs ◽

Infected Fish ◽

Control Group ◽

Fiber Glass ◽

Koi Herpesvirus ◽

Specific Immunity ◽

Effective Period

Penelitian dengan tujuan untuk mengetahui masa induksi kekebalan spesifik hingga level protektif terhadap KHV pada populasi ikan mas yang diinfeksi secara buatan melalui teknik pemaparan terkontrol telah dilakukan pada skala laboratorium. Pemaparan terkontrol terhadap ikan positif KHV dilakukan selama 3 hari pada suhu 24oC-26oC, selanjutnya ikan dipindahkan ke wadah volume 300 liter yang diisi ikan uji sebanyak 200 ekor/wadah dan suhu air berkisar antara 31oC-34oC. Perlakuan yang diterapkan adalah periode induksi: (A) periode induksi selama 1 minggu dari akhir masa pemaparan terhadap KHV, (B) periode induksi selama 2 minggu dari akhir masa pemaparan terhadap KHV, (C) periode induksi selama 3 minggu dari akhir masa pemaparan terhadap KHV, (D) periode induksi selama 4 minggu dari akhir masa pemaparan terhadap KHV, dan (E) tanpa periode induksi setelah dilakukan pemaparan terhadap KHV. Pada hari ke-21, seluruh kelompok perlakuan diinfeksi KHV secara buatan dengan teknik kohabitasi. Pengamatan dilakukan terhadap tingkah laku, gejala klinis, dan mortalitas ikan uji yang dilakukan setiap hari hingga akhir percobaan. Rataan persentase sintasan ikan uji tertinggi pada akhir riset diperoleh pada periode induksi selama 3 minggu sebesar 53,75%; diikuti oleh periode induksi selama 2 minggu sebesar 33,75%; selama 1 minggu sebesar 18,75%; selama 4 minggu sebesar 12,5%; dan kelompok kontrol sebesar 7,5%.The research with the aim to study an effective period to develop specific immunity against Koi Herpesvirus on common carp population which was obtained by cohabitation technique had been done in laboratory scale. Cohabitation of KHV infected fish at 24oC-26oC was for 3 days, and then the fish was moved to fiber glass tank at 31oC-34oC to develop specific immunity. The treatments applied in the research were: (A) a week period induction, (B) two weeks period induction, (C) three weeks period induction, (D) four weeks period induction, and (E) without period induction as a control group. Fish test were challenge to KHV infection at the end of each defined period induction by cohabitation method for 2 weeks lasting. Examination on behavior, clinical signs, and mortality of fish test were taken place daily. The results showed that the highest survival rate was found on three weeks period induction (53.75%), and followed by two weeks period induction (33.75%), one week period induction (18.75%), four weeks period induction (12.5%), and the lowest was found on the control (7.5%).

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Potency of Garlic Extract Against Koi Herpesvirus (KHV) in Common Carp

Jurnal Akuakultur Indonesia ◽

10.19027/jai.6.147-154 ◽

2007 ◽

Vol 6 (2) ◽

pp. 147

Author(s):

Sri Nuryati ◽

D. Puspitaningtyas ◽

D. Wahjuningrum

Keyword(s):

Survival Rate ◽

Common Carp ◽

Clinical Symptoms ◽

Hemoglobin Concentration ◽

Blood Parameters ◽

Garlic Extract ◽

Virus Suspension ◽

Cell Numbers ◽

Koi Herpesvirus ◽

Leukocyte Number

Prevention of koi herpesvirus (KHV) infection using chemicals or medicines was ineffective way. In this study garlic extract was used to prevent KHV infection. Virus suspension of 0.1 ml and garlic extract of 0.1 ml in different dosage, i.e., 100, 200 and 300 ppt, was injected into common carp body. Hemoglobin concentration, red and white blood cell numbers, and leukocyte number were counted. The results of study showed that administration of 300 ppt of garlic extract could produce higher survival rate (67.5%), good blood parameters and clinical symptoms compared to other treatments. Keywords: garlic, KHV, common carp ABSTRAK Upaya penanggulangan wabah Koi Herpesvirus (KHV) menggunakan bahan-bahan kimia atau obat-obatan adalah tidak efektif. Pada penelitian ini dilakukan pemberian ekstrak bawang putih untuk menanggulangi infeksi KHV. Suspensi virus sebanyak 0,1 ml ditambahkan dengan 0,1 ml ekstrak bawang putih dengan berbagai konsentrasi, yaitu 100, 200 dan 300 ppt, disuntikkan ke dalam tubuh ikan mas. Kadar hemoglobin, jumlah sel darah merah dan sel darah putih jenis dan jumlah leukosit diamati. Hasil penelitian menunjukkan bahwa pemberian ekstrak bawang putih sebanyak 300 ppt menghasilkan kelangsungan hidup (67,5%) yang lebih baik dibandingkan dengan perlakuan lainnya, dan begitu pula dengan gambaran darah serta gejala klinisnya. Kata kunci: bawang putih, KHV, ikan mas

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First Detection of Koi Herpesvirus in Cultured Common Carp Cyprinus carpio in Japan

Fish Pathology ◽

10.3147/jsfp.39.165 ◽

2004 ◽

Vol 39 (3) ◽

pp. 165-167 ◽

Cited By ~ 83

Author(s):

Motohiko Sano ◽

Takafumi Ito ◽

Jun Kurita ◽

Takanori Yanai ◽

Naoki Watanabe ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Koi Herpesvirus ◽

Common Carp Cyprinus Carpio ◽

Carp Cyprinus Carpio

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Effication of Viral Nervous Necrosis DNA as Vaccines for Cromileptes altivelis

Indonesian Journal of Tropical Aquatic ◽

10.22219/ijota.v3i2.13891 ◽

2020 ◽

Vol 3 (2) ◽

Author(s):

Wiwien Mukti Andriyani ◽

Sri Murtini ◽

Alimuddin Alimuddin

Keyword(s):

Dna Vaccine ◽

Challenge Test ◽

Virus Titer ◽

Dna Vaccination ◽

Post Vaccination ◽

Viral Nervous Necrosis ◽

Antibody Induction ◽

Actin Promoter ◽

Two Stages ◽

Made In

Viral nervous necrosis (VNN) is a disease that often infects groupers. It has caused mass death in more than 34 species of marine fish. DNA vaccination might become a solution againts the infection. The construction of the pmBA-CP DNA vaccine consisting of the beta-actin promoter of medaka fish (Oryzias latipes) and capsid protein (CP) encoding VNN RNA2 has been made in previous studies. This study aimed to test the efficacy of the pmBA-CP DNA vaccine for VNN. The experiment consisted of two stages, namely (1) detection of anti-VNN antibody induction in vaccinated fish using ELISA, and (2) challenge test for fish vaccinated with the VNN virus. Grouper (body length 8 cm to 10 cm) were divided into two groups with a density of 5 fish 60 L–1. The fish in the first group were vaccinated with pmBA-CP intramuscularly at a dose of 12.5 µg per fish, while the second group of fish were not vaccinated. Antibody titer testing was carried out before treatment, and 1 d, 7 d, 14 d, 21 d, 28 d, and 35 d after vaccination. The challenge test was carried out on the 60th day after vaccination. The results showed that the S / P ratio in the vaccinated fish serum was higher than unvaccinated fish at 21 d to 35 d post-vaccination. DNA vaccination was able to induce anti-VNN antibodies of grouper. The results of the challenge test for vaccinated fish using VNN virus titer 103.5 FID50/0.2 mL showed 60% of survival rate. Thus, the pmBA-CP DNA vaccine could be useful for increasing grouper immunity, and support production of grouper.

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