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Rapid method for identification of transgenic fish zygosity Alimuddin, .; Yoshizaki, G.; Carman, O.
Jurnal Akuakultur Indonesia Vol. 6 No. 2 (2007): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (178.956 KB) | DOI: 10.19027/jai.6.177-182

Abstract

Identification of zygosity in transgenik fish is normally achieved by PCR analysis with genomic DNA template extracted from the tissue of progenies which are derived by mating the transgenic fish and wild-type counterpart.  This method needs relatively large amounts of fish material and is time- and labor-intensive. New approaches addressing this problem could be of great help for fish biotechnologists.  In this experiment, we applied a quantitative real-time PCR (qr-PCR) method to analyze zygosity in a stable line of transgenic zebrafish (Danio rerio) carrying masu salmon, Oncorhynchus masou D6-desaturase-like gene. The qr-PCR was performed using iQ SYBR Green Supermix in the iCycler iQ Real-time PCR Detection System (Bio-Rad Laboratories, USA).  Data were analyzed using the comparative cycle threshold method.  The results demonstrated a clear-cut identification of all transgenic fish (n=20) classified as a homozygous or heterozygous.  Mating of those fish with wild-type had revealed transgene transmission to the offspring following expected Mendelian laws. Thus, we found that the qTR-PCR to be effective for a rapid and precise determination of zygosity in transgenic fish. This technique could be useful in the establishment of breeding programs for mass transgenic fish production and in experiments in which zygosity effect could have a functional impact. Keywords: quantitative real-time PCR; zygosity; transgenic fish; mass production   ABSTRAK Identifikasi sigositas ikan transgenik biasanya dilakukan menggunakan analisa PCR dengan cetakan DNA genomik yang diekstraksi dari jaringan ikan hasil persilangan antara ikan transgenik dan ikan normal.   Metode ini memerlukan ikan dalam jumlah yang banyak, dan juga waktu serta tenaga.  Pendekatan baru untuk mengatasi masalah tersebut akan memberikan manfaat besar kepada peneliti bioteknologi perikanan.  Pada penelitian ini, kami menggunakan metode PCR real-time kuantitatif (krt-PCR) untuk menganalisa sigositas pada satu strain ikan zebra (Danio rerio) transgenik yang membawa gen D6-desaturase-like dari ikan salmon masu, Oncorhynchus masou.  krt-PCR dilakukan menggunakan iQ SYBR Green Supermix pada mesin iCycler iQ Real-time PCR Detection system (Bio-Rad Laboratories, USA).  Data dianalisis menggunakan metode pembandingan nilai cycle threshold.  Hasil penelitian menunjukkan bahwa semua ikan transgenik (n=20) yang diidentifikasi dapat diklasifikasikan secara jelas sebagai ikan homosigot atau heterosigot.  Persilangan antara ikan transgenik tersebut dengan ikan normal menunjukkan transmisi transgen ke keturunannya mengikuti hukum segregasi Mendel.  Dengan demikian, metode krt-PCR adalah efektif untuk penentuan sigositas secara cepat dan tepat pada ikan transgenik.  Teknik ini dapat berguna dalam program produksi ikan transgenik secara massal dan dalam percobaan dimana faktor sigositas memberikan pengaruh nyata. Kata kunci: kuantitatif real-time PCR; sigositas, ikan transgenik; produksi massal
Oral Administration of 17α-Methyltestosterone Increased Male Percentage of Freshwater Crayfish Cherax quadricarinatus Carman, O.; Jamal, M.Y.; Alimuddin, .
Jurnal Akuakultur Indonesia Vol. 7 No. 1 (2008): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (278.216 KB) | DOI: 10.19027/jai.7.25-32

Abstract

Cherax quadricarinatus is one of freshwater crayfish species that has enormous potential for expanding its farming in future.  Application of monosex male culture using steroid sex hormone administration method during the period of sex differentiation or early developmental stage might be increased efficiency in farming.  This study was aimed to increase male of C. quadricarinatus by oral administration of diet containing 17α-methyltestosterone (MT) towards production efficiency.  Two-week-old of Cherax quadricarinatus were fed ad libitum on diets containing various dose of MT, i.e., 25, 50, 75, 100 and 150 mg/kg diet or diet containing no MT as control, 3 times daily for 30 days.  After MT-treatment, crayfish were fed frozen Chironomus sp.  and shrimp diet.  Sex ratio, survival and growth rate (by length and weight) were observed at the end of experiment.  Sex was determined by visual observation; the male sex organ is located at the fifth walking leg while the female is at the third.  Data was analyzed by F and BNT tests.  The results of study show that administration of MT was significantly changed the male ratio of crayfish.  Treatment dose of 50 mg/kg diet was effective to increase male sex percentage from 24.93% (control) to be 59.96%. Growth was also significantly being improved, while survival rate was insignificant.  Thus, oral administration of MT is an effective way to increase male sex percentage of crayfish, although other methods and the time of hormone administration are needed to be verified to obtain maximal results. Keywords: monosex, 17α-methyltestosterone, sex reversal, Cherax quadricarinatus   ABSTRAK Salah satu jenis lobster air tawar yang berpotensi tinggi untuk dikembangkan usaha budidayanya adalah Cherax quadricarinatus. Aplikasi teknik budidaya tunggal kelamin (monoseks) dengan metode pemberian hormon seks steroid yang diberikan pada saat diferensiasi kelamin atau masa perkembangan awal ikan diduga dapat meningkatkan efisiensi usaha.  Penelitian ini ditujukan untuk meningkatkan persentase C. quadricarinatus jantan menggunakan metode seks reversal melalui pemberian pakan yang mengandung 17α-metiltestosteron (MT) sebagai upaya efisiensi produksi.  C. quadricarinatus umur 2 minggu diberi pakan yang mengandung MT dengan dosis 25, 50, 75, 100 dan 150 mg/kg pakan atau tanpa hormon secara ad libitum, 3 kali sehari selama 30 hari.  Setelah perlakuan lobster uji diberi pakan alami Chironomus sp. beku dan pakan udang. Parameter yang diamati meliputi nisbah kelamin, kelangsungan hidup dan pertumbuhannya (panjang dan berat mutlak), yang dilakukan pada akhir penelitian. Identifikasi jenis kelamin dilakukan secara visual; alat kelamin lobster jantan terdapat pada bagian pangkal kaki jalan kelima, yang betina terletak pada bagian dasar kaki jalan ketiga.  Data dianalisis menggunakan uji F dan BNT. Hasil penelitian menunjukkan bahwa pemberian hormon MT berpengaruh nyata terhadap persentase kelamin jantan lobster. Perlakuan dengan dosis 50 mg/kg pakan efektif untuk meningkatkan persentase jantan C. quadricarinatus dari 24,93% (kontrol) menjadi 59,96%.  Pertumbuhan panjang dan berat mutlak  juga menunjukkan pengaruh yang berbeda nyata, sementara kelangsungan hidup tidak berbeda.  Dengan demikian pemberian hormon MT melalui pakan cukup efektif untuk meningkatkan persentase lobster jantan yang dihasilkan, meskipun penggunaan metode lain dan waktu pemberian hormon MT masih perlu diteliti untuk memperoleh hasil maksimal.  Kata kunci: tunggal kelamin, 17α-metiltestosteron, seks reversal, Cherax quadricarinatus
Characterization of β-Actin Promoter from Nile Tilapia (Oreochromis niloticus) Alimuddin, .; Octavera, A.; Arifin, O.Z.; Sumantadinata, K.
Jurnal Akuakultur Indonesia Vol. 7 No. 2 (2008): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (450.672 KB) | DOI: 10.19027/jai.7.115-127

Abstract

Promoter is one of the factors determining the successful of transgenesis.  In this study we isolated and characterized β-actin promoter from Nile tilapia (tiBP) towards production of autotransgenic tilapia.  β-actin promoter has high activity in muscle.  Sequence of tiBP promoter was isolated by using PCR method. Sequencing was performed using ABI PRISM 3100 machine. Analysis of sequences was conducted using GENETYX version 7 and TFBind softwares. DNA fragment of PCR amplification product digested from the vector cloning was then ligated with pEGFP-N1 to generate ptiBP-EGFP construct. The construct was microinjected into one-cell stage of zebrafish (Danio rerio) embryos to test the tiBP promoter activity. EGFP gene expression was observed by fluorescence microscope.  The result of sequence analysis showed that the length of DNA fragment obtained is about 1.5 kb and containing the evolutionary conserved sequences of transcription factor for β-actin promoter including CCAAT, CArG and TATA boxes.  Furthermore, tiBP sequence in ptiBP-EGFP construct could regulated GFP expression in muscle of zebrafish embryos injected with the construct. The results suggested that PCR amplification product is the regulator sequence of tilapia β-actin gene. Autotransgenic tilapia can be then produced by changing GFP gene fragment of ptiBP-EGFP construct with genes from tilapia encoding important traits in aquaculture. Keywords:  cloning, β-actin promoter, autotransgenic, EGFP, Oreochromis niloticus, Danio rerio   ABSTRAK Promoter merupakan salah satu faktor penentu keberhasilan transgenesis.  Pada penelitian ini kami mengisolasi dan mengkarakterisasi promoter β-actin dari ikan nila (tiBP) dalam rangka pembuatan ikan nila autotransgenik. Promoter β-actin memiliki aktivitas tinggi pada jaringan otot. Sekuens promoter tiBP diisolasi menggunakan metode PCR.  Sekuensing dilakukan menggunakan mesin ABI PRISM 3100. Analisa sekuens menggunakan software GENETYX versi 7 dan TFBind.  Fragment DNA hasil amplifikasi PCR yang didigesti dari vektor kloning selanjutnya diligasi dengan pEGFP-N1 untuk membuat konstruksi ptiBP-EGFP. Konstruksi ptiBP-EGFP dimikroinjeksi ke embrio ikan zebra (Danio rerio) fase 1 sel untuk menguji aktivitas promoter tiBP. Ekspresi gen EGFP diamati menggunakan mikroskop fluoresens. Analisa sekuens menunjukkan bahwa panjang fragmen DNA hasil amplifikasi PCR sekitar 1,5 kb dan memiliki faktor transkripsi yang konserf untuk promoter β-actin, yaitu CCAAT, boks CArG dan TATA.  Selanjutnya, sekuens tiBP dalam konstruksi ptiBP-EGFP mampu mengendalikan ekspresi gen EGFP pada jaringan otot embrio ikan zebra yang dimikroinjeksi dengan konstruksi tersebut.  Dengan demikian dapat disimpulkan bahwa fragmen DNA hasil amplifikasi PCR tersebut merupakan sekuens promoter β-actin ikan nila. Pembuatan ikan nila autotransgenik selanjutnya dapat dilakukan dengan mengganti gen EGFP pada pktBA-EGFP dengan gen-gen asal ikan nila yang mengkodekan karakter penting dalam budidaya ikan. Kata kunci:  kloning, promoter β-actin, autotransgenik, EGFP, Oreochromis niloticus, Danio rerio
Hematology of common carp following DNA vaccination and koi herpesvirus challenge test Nuryati, Sri; Maswan, N.A.; Alimuddin, .; Sukenda, .; Sumantadinata, K.; Pasaribu, F.H.; Soejoedono, R.D.; Santika, A.
Jurnal Akuakultur Indonesia Vol. 9 No. 1 (2010): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (194.781 KB) | DOI: 10.19027/jai.9.9-15

Abstract

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
Effectiveness of B-actin promoter on driving target gene expression in common carp transgenesis Hidayani, Andi Aliah; Carman, Odang; Alimuddin, .
Jurnal Akuakultur Indonesia Vol. 10 No. 1 (2011): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (925.474 KB) | DOI: 10.19027/jai.10.16-23

Abstract

Promoter in transgene construct plays an important role on regulating of transgene expression level in transgenic fish. In fish transgenesis, researcher convinced that use all-fish gene construct is safety and prospective. This study was performed to compare effectiveness b-actin promoter, - the promoter which has ubiquitous, constitutive, housekeeping characteristics, from common carp (homologous) and from tilapia and medaka b-actin promoters (heterologous) in driving of green fluorescent protein (GFP) expression as a model of target gene on common carp transgenesis. These gene constructs were separately microinjected into cytoplasm of 60 one-cell-stage common carp embryos. The results suggested that 70% survival rate at embryo stage and 45% hatching rate values showed that the microinjection was performed successfully. Percentage of embryos expressing GFP gene were slightly higher when injected using common carp and medaka promoters than those of using tilapia promoter. Percentage of larvae expressing GFP using common carp promoter was similar with medaka promoter. Furthermore, GFP expression using common carp b-actin promoter could be detected at one-week-old larvae, while GFP expressing using medaka b-actin promoter was lasted at 2-day-old larvae. The results demonstrated that homologous promoter more effective in driving of a target gene expression than that of heterologous promoter.  Key words: homologous promoter, GFP, transgenesis, common carp   ABSTRAK Promoter dalam konstruksi transgen berperan penting dalam pengaturan tingkat ekspresi transgen pada ikan transgenik. Dalam transgenesis ikan, peneliti meyakini bahwa penggunaan konstruksi gen "all-fish" adalah aman dan prospektif.  Penelitian ini dilakukan untuk membandingkan efektivitas promoter β-aktin, - promoter yang memiliki ciri ubiquitous, constitutive, dan housekeeping, dari ikan dari ikan mas (homolog) dan ikan nila dan ikan medaka (heterolog) dalam mengendalikan ekspresi gen GFP sebagai model gen pada transgenesis ikan mas. Setiap  konstruksi gen tersebut diinjeksikan secara terpisah ke sitoplasma embrio ikan mas fase 1 sel sebanyak 60 embrio. Hasil penelitian dengan kelangsungan hidup embrio 70% dan derajat penetasan 45% menunjukkan bahwa kegiatan mikroinjeksi berhasil dengan baik.  Persentase embrio mengekspresikan gen GFP yang diinjeksi konstruksi gen dengan promoter β-aktin ikan mas dan ikan medaka sedikit lebih tinggi dibandingkan dengan yang menggunakan promoter β-aktin ikan nila.  Selanjutnya, ekspresi gen GFP yang dikendalikan oleh promoter β-aktin ikan mas dapat dideteksi pada larva berumur 1 minggu, sedangkan ekspresi GFP dengan promoter β-aktin ikan medaka hanya bisa terdeteksi hingga larva berumur 2 hari.  Hasil penelitian menunjukkan bahwa promoter homolog adalah lebih efektif dalam mengatur ekspresi gen target dibandingkan dengan promoter heterolog. Kata kunci: promoter homolog, GFP, transgenesis, ikan mas
Production and bioactivity test of recombinant protein common carp growth hormone Utomo, Deny Sapto Chondro; Alimuddin, .; Sudrajat, Agus Oman; Faizal, Irvan
Jurnal Akuakultur Indonesia Vol. 10 No. 1 (2011): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1361.474 KB) | DOI: 10.19027/jai.10.44-50

Abstract

This study aimed to produce recombinant growth hormone (rGH) protein of common carp (Cyprinus carpio) and evaluate its bioactivity. DNA fragment encoding mature GH protein of common carp (mCcGH) was amplified by polymerase chain reaction (PCR) method and PCR products were then ligated into pCold I to generate pCold I-mCcGH protein expression vector. Escherichia coli BL21 (DE3) harboring pCold I-mCcGH was cultured in the 2xYT medium at 15 °C for 24 hours and protein production was induced by isopropyl-beta-thio galactopyranoside (IPTG). The inclusion bodies containing rGH protein from E. coli transformants were isolated by sonication method and analyzed by SDS-PAGE. The result showed that rGH with molecular weight of about 25 kDa was obtained. Common carp juveniles with average body weight of 5.2±0.4 g were intramuscularly injected once a week for 4 weeks with rGH protein solution from 1 μg bacterial cells per gram fish body weight. The result showed that juveniles fish injected with rGH grew 106.56% higher than control. This result indicated that rGH produced in E. coli BL21 possessed biological activity and it may be useful to improve growth of aquaculture species.Key words: growth hormone, recombinant protein, common carp ABSTRAKPenelitian ini bertujuan menghasilkan protein rekombinan hormon pertumbuhan (growth hormone, GH) dari ikan mas (Cyprinus carpio) dan menguji bioaktivitasnya. Fragmen DNA penyandi protein matang (mature) GH ikan mas (mCcGH) diamplifikasi dengan menggunakan metode PCR dan hasilnya kemudian diligasi ke dalam pCold-I untuk menghasilkan konstruksi vektor ekspresi pCold-I-mCcGH. Plasmid pCold-I-mCcGH ditransformasi ke bakteri Escherichia coli BL21 (DE3), dikultur dalam media 2xYT cair pada suhu 15°C selama 24 jam dan produksi protein diinduksi dengan menggunakan isopropyl-beta-thio galactopyranoside (IPTG). Badan inklusi yang mengandung protein rekombinan GH (rGH) dari bakteri E. coli transforman diisolasi menggunakan metode sonikasi dan dianalisis dengan menggunakan SDS-PAGE. Hasil penelitian menunjukkan bahwa rGH dengan bobot molekul sekitar 25 kDa berhasil diproduksi. Benih ikan mas dengan bobot rata-rata 5,15±0,4 g diinjeksi secara intramuskular satu kali per minggu selama 4 minggu dengan larutan rGH hasil ekstraksi dari 1 µg pelet bakteri/g bobot ikan. Benih yang disuntik dengan rGH tumbuh sekitar 100% lebih cepat bila dibandingkan dengan kontrol yang tidak diinjeksi rGH. Hasil ini mengindikasikan bahwa rGH yang diproduksi dalam bakteri E. coli memiliki bioaktivitas dan dapat bermanfaat untuk memacu pertumbuhan spesies ikan-ikan budidaya.Kata kunci: hormon pertumbuhan, protein rekombinan, ikan mas
Performance of Nile tilapia juvenile fed diet containing different dosages ofrecombinant common carpgrowth hormone Hardiantho, Dian; Alimuddin, .; Prasetiyo, Arief Eko; Yanti, Dwi Hany; Sumantadinata, Komar
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 (876.198 KB) | DOI: 10.19027/jai.11.17-22

Abstract

The studywas conductedtodetermine thedose of recombinant common carp growth hormone (rCcGH) supplemented in the commercial diet that generates the best performanceonbodyweight, biomass, andfeed conversion rate of juvenile tilapia.The dose of rCcGH administered was10, 20, and30mg/kg of commercial feed, andnorGH supplementation as control. Eachtreatment was designed in triplicates. Juveniles at average bodylengthof about2cm(average body weight of 0.7 g) were reared in a density of25fish/m2, for three weeks in hapa net at a size of 2×2×1 m3. Fish were fed diet containing rCcGH twicea week with an interval ofthreedays. The juvenile were fed with the rCcGH supplemented diet three times daily at satiation. The results showedthat average body weight andbiomass of fish treated by rCcGH with the doses of 20 and 30 mg/kg feed was higher (p<0.05) than that of 10 mg/kg and control. In addition, feed conversionrate of 20 and 30 mg/kg feed treatments was lower (p<0.05) than that of 10 mg/kg and control. RT-PCR analysisshowedthat expression level of IGF-1 gene in juvenile liver was increased in parallel with the increasing of rCcGH dosages supplemented in diet.This suggestedthatIGF-1 plays an important rolein the induction ofgrowth,andfeed conversionefficiencyof tilapiafed diet containing rCcGH. The bestperformance of juvenile can be obtainedby feeding with diet containingrCcGH20‒30mg/kgof feed. Keywords: rCcGH, IGF-I, recombinant growthhormone,performance, Nile tilapia.
Growth and survival of giant gourami juvenile immerse indifferent frequencies using recombinant growth hormone Syazili, Aras; Irmawati, .; Alimuddin, .; Sumantadinata, Komar
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 (786.856 KB) | DOI: 10.19027/jai.11.23-27

Abstract

This study was aimed to determine the immersion frequency of recombinant growth hormone (rGH) in the growth stimulation of giant gourami juvenile. A total of 50 larvae at second day after first feeding, was immersed in 30 ppt salt water for 2 minutes, and then transferred into 200 mL of 9 ppt salt water containing 30 mg/L rGH in inclusion bodies form. Immersion was performed for 60 minutes and repeated once a week for two, three, four, and five weeks. As the control, fish was immersed with inclusion bodies without rGH (control-1), bovine serum albumin (control-2), 9ppt saline water without rGH (control-3) and in freshwater (control-4). Fish were kept in 7L aquaria during the first four weeks, transferred into 50L aquaria for two weeks, and then in hapa (2×1×1 m3 in size) for two weeks. The results showed that the highest (p<0.05) average body weight of giant gourami juvenile (2.23±0.19 g) was obtained at four times (four weeks) immersion treatment. Similar body weight of fish in two times (1.72±0.29 g), three times (1.68±0.34 g), and five times (1.81±0.17 g) immersion was obtained. Fish body weight of all controls was lower than the rGH-immersed fish. Average body weight of four time rGH-immersed fish (4.01 g) remained higher than other treatment and control (1.96 to 2.36 g) on 8th week of fish rearing. Survival of four times (96.67±1.15%), and five times rGH-immersion fish (95.33±4.16%) was similar, and higher than other treatments and controls (77.33 to 89.33%). Thus, higher growth rate and survival of giant gourami juvenile could be achieved by four times rGH immersion. Keywords: recombinant protein, growth hormone, immersion frequency, giant gourami
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
Growth, feed conversion and retention, and proximate of eel juvenile treated by immersion of recombinant giant grouper growth hormone Handoyo, Boyun; Alimuddin, .; Utomo, Nur Bambang Priyo
Jurnal Akuakultur Indonesia Vol. 11 No. 2 (2012): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (929.363 KB) | DOI: 10.19027/jai.11.132-140

Abstract

This study was aimed to determine the immersion dose of recombinant giant grouper growth hormone (rElGH) to increase growth of eel juvenile (glass eel). After shock salinity treatment (NaCl 3% for 2 min), glass eel were immersed in water containing 0.9% NaCl, 0.01% bovine serum albumin, and different of dose of rElGH (0, 0.12, 1.2, 12 and 120 mg/L). Glass eel were fed on blood worm for the first month, and commercial diet for the second month of rearing. Fish rearing was performed in 60 L glass aquarium at density of 150 fish per aquarium. The results showed that higher in growth body weight was obtained in immersion dose of 12 mg/L, by increment of about 37.4% higher compared to that of control. The immersion dose of 12 mg/L also increased specific growth rate by 29.3% higher (p<0.05) compared to control. Furthermore, at that immersion treatment, feed conversion ratio of blood worm and commercial diet decreased by 33.7% and 25.6% compared to control, respectively. Protein (7.15±0.08%) and lipid (9.95±0.10%) retentions in 12 mg/L rElGH-treated fish were higher (p<0.05) than those of control (protein 6.17±0.07%; lipid 5.73±0.06%). Lower in protein content (12.73%), while higher content in lipid (8.35%) and crude carbohydrate (3.22%) were found in 12 mg/L rElGH-treated fish compared to those of control (13.24%, 5.90%, and 1.76%, respectively). Thus, in general, rElGH immersion dose of 12 mg/L could generate high performances of eel juvenile, and application of rElGH can be useful to increase aquaculture eel production.Keywords: immersion dose, growth hormone, growth, glass eel
Co-Authors . Irmawati . Kurdianto . Suharsono A. Octavera A. Santika Abdul Halim Talli Abyadul Fitriyah Ade Sunarma Adi Sucipto, Adi Adinda Rizky Tasrianty Afdal Madein Afif Abdurrahman Afpriyaningrum MD Agus Oman Sudrajat Agus Priyadi Ahmad Faisal Ahmad Muzaki Ahmad Ramzi Ghulam Syam Ahmad Zaki Alfiah Mutmainnah Ali Alyaminy, Humaidatul Aminah Zaim Aman Sentosa Panggabean Amin, Muhmmadiyah Andi Aliah Hidayani Andi Haerul Andi Parenrengi Andi Parenrengi Andi Parenrengi Andi Tenriulo Andi, K. Petta Lolo, Arjuni Andi, Maulana K. Angela Mariana Lusiastuti Anggriani, Emi Anhar ANNA OCTAVERA Anny Hary Ayu Apriwanto, Indra Aras Syazili Arief Boediono Arief Eko Prasetiyo Arien Heryansyah Asdar Asep Permana Asep Sopian Asri Ainun Amaliah Aulia Saputra Awi Dassa Ayi Santika Azis Ali Baderiah Baderiah Bahri, Nur Alim Benyamin Timotiwu, Paul Bohari Yusuf Boyun Handoyo Budimawan Chaeroni, Ahmad Chaerunnisa Chotban, Sippah DARMAWAN SETIA BUDI Darussalam Darwin, Khadijah DEDI JUSADI Delfian Masrura Deny Sapto Chondro Utomo Desy Nurul Astuti Devi Wardan Sari Dewi Yuniati Dian Hardiantho Dian Hardiantho Diki Nurbaldatun Islam Dinamella Wahjuningrum Dinar Tri Soelistyowati Djadir Dwi Hany Yanti Eko Pramono Emma Suryati Emma Suryati Enang Harris Enang Harris Endi Permata Eni Kusrini Eni Kusrini Erma Primanita Hayuningtyas F.H. Pasaribu Fachruddin Fachruddin Fadhila Maharani Putri Fajar Anggraeni Fatihah, Anariski Fatimah Akbal Fauzan Wahib Alsani Fauziah Zainuddin Firman Firmansyah Fitriadi Fitriyah Husnul Khotimah Fitry Hasdanita Fredinan Yulianda G. Yoshizaki GEMI TRIASTUTIK Gloria Ika Satriani GORO YOSHIZAKI Goro Yoshizaki Gunawan Gunawan Gustikasari Amin H. Arfah Hadi Nasbey Hamda Hamid Habbe Hamzah Hanipa Hardiantho D Harmayani, Ria Harton Arfah Hary Krettiawan Haryanti Haryanti Hasan Nasrullah Hassan, Muhammad Nasiru Herjayanto, Muh. Hisyam Ihsan Ibnu Izzah Idrus Sahar Muhammad Iis Diatin Ika Kartika Ikhwanul Arifan ILHAM Ilham Minggi Imron Imron Indah Mastuti Irma Andriani IRMA ANDRIYANI Irmawati, Budimawan Irvan Faizal Irvan Faizal IRVAN FAIZAL Irwan Irzal Effendi Ita Djuwita Jasmadi Jasmadi Jean-Christophe Avarre Joko Santoso Jubaedah Julie Ekasari Jumrah123 K. Chaitanya Deepti K. Sumantadinata Kadriani, Nur Annisa Kamaruddin Kartini, Selvira Febriani Katarina Manik, Tumiar Ketut Mahardika Ketut Sugama Khaerullah khairuzzazi Komar Sumantadinata Komar Sumantadinata Komar Sumantadinata KOMAR SUMANTADINATA Leman, Swanggi Lianasari, Ika Yekti Lidyasari Liza Luky Adrianto M. SASMITO JATI M. Zairin Junior M.Y. Jamal Madein, Afdal Maharani, Adinda Marwah, Andi Marwah Mashud MEDIATY Megawati, Novi Mia Setiawati Miftahul Jannah Mira Mawardi MMSI Irfan ,S. Kom Mochamad Syaifudin Muh Alias L. Rajamuddin Muh. Akmal, Andi Muh. Alias L. Rajamuddin MUHAMMAD AGUS SUPRAYUDI Muhammad Ammar Naufal Muhammad Awal Ramadhan Muhammad Firdaus Muhammad Ikhsan Muhammad Jufri Muhammad Muttaqin Muhammad Sabir Muhammad Safir Muhammad Wawan Muhammad Zairin Jr. Muhammad Zairin Jr. Muhammadiyah Amin MUNTI YUHANA Muslim Muslim Mustika Sari, Mustika Musyfikah Ilyas Muzahar N.A. Maswan Nadhirah Nagu Nagu, Nadhirah Nasrullah Nenima Halawa Nining Asniar Ridzal Noerlitasari Nugraha, Adrianto Fahrizal Nunak Nafiqoh Nur Bambang Priyo Utomo Nur Bambang Priyo Utomo Nur, Rezki Amaliah Nurdin Arsyad Nurfadilla Nurhalija Nurhayati Nurhidayat, Muhammad Nurlaelah Nurlaelah Syarifuddin Nurly Faridah Nurly Faridah Nurul Fadhilah Nurwati Djam’an O. Carman O.Z. Arifin Odang Carman Oman Unju Subandi P. Ratna Kumar Permata Sari, Anggun Petrus Rani Pong-Masak Priatama, Rizki Purwaningsih, Yuli Pustika Ratnawati Putri, Angelina Atika R Rifai R.D. Soejoedono Rahmi, Kurnia Anggraini Ratu Siti Aliah Ratu Siti Aliah RATU SITI ALIAH Reni Agustina Lubis Ridwan RIDWAN AFFANDI Risfiyanty, Dwi Kartika Riswandi Rizky Ayu Fatimah Ridwan Rodliyah Rohayu, Rina Rosidah Rusli RUSTIDJA S. Mariam Samsu Adi Rahman Sani, NurHaslina Sanusi Saputra, Ade Maulana Sari , Nur Indah Sari Budi Moria Sembiring Sarmila Sekar Sulistyaning Hadiwibowo Setiawati M Shabarullah Shalsyabila , Zabina Shandi Irawan Sholeh Ridwan Siti Aslamyah Siti Fadilah Sitti Fakhriyyah SLAMET SUBYAKTO Soelistyowati DT Soerja Koesnarpadi Soko Nuswantoro Sri Khayati sri murtini . Sri Nuryati Sri Nuryati Srihartini, Yusi St. Fatmawati. L St. Hidayah Triana St. Mukarramah Sukenda . Sukenda Sukenda Sukmawati Ali Sumardin Raupu Sundari Sutamrin Syafrianto, Syafrianto Syuhada Restu Danupratama T. Takeuchi Tahrim, Tasdin Talib, Dr. Ahmad Tasdin Tahrim Tedy Murtedjo Tri Setyo Guntoro Usman Mulbar Utut Widyastuti Utut Widyastuti Utut Widyastuti Utut Widyastuti Wa Ode NurRadhiah Ridjalu Wahidah Sanusi Wartono Hadie Wicaksono, Aryo Wenang WIDANARNI WIDANARNI Y. Hadiroseyani Yulintine Yulintine Yuni Puji Hastuti Yuningsih, Andri Yusra, Andi Zairin Jr M Zakki Zainun