Indonesian eel (Anguilla bicolor bicolor) has recently been cultivated intensively in Indonesia. The growth of Indonesian eel is slow. This study was performed to determine a proper delivery method of recombinant giant grouper (Epi-nephelus lanceolatus) growth hormone (rElGH) and observe aquaculture and nutrient related performances of Indonesian eel elver after rElGH treatment. Three delivery methods of rElGH, namely immersion, orally, and combination of immersion and orally were tested. The dosage of rElGH by immersion (12 mg L"1 water) and orally administration (30 mg kg-1 feed) were obtained in previous studies. Fish were maintained in 23 L glass aquaria for 120 days, at density of 45 fish in each aquarium. The results showed that higher (P<0.05) biomass of harvest (73.68±2.07 g) was obtained in immersion and oral combination treatment, by increment of about 102.9% compared to control (36.32±0.97 g). Administration of rElGH also improved protein and lipid retentions by 142.5%, and 720.0% compared to those of control, respectively (P<0.05). In addition, rElGH treatment increased appetite, while artificial feed conversion ratio (4.75) was 55.15% lower (P<0.05) compared to that of control (7.37). Combination of rElGH immersion and oral administrations also increased hepatosomatic index, and insuline-like growth factor-1 gene expression level. Thus, rElGH administration via immersion and oral combination was a proper method to improve growth of eel elver, and application of rElGH can be useful to increase aquaculture production. Abstrak Ikan sidat (Anguilla bicolor bicolor) telah dibudidayakan secara intensif di Indonesia. Pertumbuhan ikan sidat relatif lambat. Penelitian ini dilakukan untuk menentukan metode pemberian hormon pertumbuhan rekombinan ikan kerapu kertang (Epinephelus lanceolatus) (rElGH) untuk memacu pertumbuhan, dan mengevaluasi performa budi daya dan pemanfaatan pakan pada elver ikan sidat. Tiga metode pemberian rElGH diuji, yakni perendaman, oral, dan kombinasi perendaman dan oral. Dosis rElGH yang digunakan merupakan hasil penelitian sebelumnya, yakni 12 mg L-1 untuk me-tode perendaman dan 30 mg kg-1 pakan untuk metode oral. Ikan dipelihara dalam akuarium volume 23 L selama 120 hari, dengan padat tebar 45 ekor per akuarium. Hasil penelitian menunjukkan bahwa biomassa tertinggi (p<0,05) diper-oleh pada perlakuan kombinasi perendaman dan oral (73,68±2,07 g), sekitar 102,9% lebih tinggi daripada kontrol tanpa pemberian rElGH (36,32±0,97 g). Perlakuan rElGH juga meningkatkan retensi protein dan lemak, masing-masing 142,5% dan 720,0% lebih tinggi daripada kontrol (p<0,05). Selanjutnya, perlakuan rElGH meningkatkan nafsu makan, dan konversi pakan (4,75) sekitar 55,2% lebih rendah (p<0,05) daripada kontrol (7,37). Perlakuan kombinasi perendaman dan oral juga meningkatkan indeks hepatosomatik, dan ekspresi gen insuline-like growth factor I. Sebagai kesimpul-an, pemberian rElGH melalui kombinasi perendaman dan oral merupakan metode terbaik dalam memacu pertumbuhan elver ikan sidat, dan penerapan rElGH dapat digunakan untuk meningkatan produksi budi daya.

In tilapia aquaculture, all male populations are preferred because they achieve higher growth rates and prevent uncontrolled reproduction. Sex reversal techniques are largely used for the control of sex in fish farming and in fundamental studies on sex determinism mechanisms. The study was conducted to determine the effect of immersion Nile tilapia larvae in water containing different honey source on male percentage and aromatase gene expression. In experiment I, a total of 30 tilapia larvae at 12 days post hatch were immersed in water containing honey derived from the forest, cultured and mangrove bees, at a dose of 10 ml L-1 for 10 hours. Fish were maintained in the same condition for two months. The results showed that percentage of male fish was similar among honey treatments (p>0.05), and they were significantly different with the control (p<0.05). In experiment II, fish were immersed in two bioactive compounds of honey, namely chrysin and potassium solution in a dose of 20 mg L-1 and 0.026 g L-1, respectively, to verify the bioactive affects sex differentiation. Aroma-g expression was analyzed by RT-PCR method. Tissue was collected at 1, 6, 12, 24 and 48 hours after immersion, and 2-month-old fish. Size fragment DNA aroma-g of female 200 bp. Chrysin and potassium immersion increased male percentage (p<0.1), this indicated that both materials were involved in Nile tilapia sex differentiation. RT-PCR analysis showed that honey, chrysin and potassium down-regulated aroma-g expression at 12 hours post immersion. Thus, honey can be used for sex reverse of Nile tilapia, and the mechanism is most likely as aromatase inhibitors. AbstrakBudi daya ikan nila dengan populasi jantan semua (monoseks) lebih memberikan keuntungan karena laju pertumbuhan-nya lebih cepat dan dapat mencegah pemijahan liar.Teknik pengarahan diferensiasi kelamin(sex reversal) digunakan untuk mengarahkan pembentukan jenis kelamin pada budi daya ikan.Penelitian ini dilakukan untuk mengevaluasi pe-ngaruh perendaman larva ikan nila menggunakan tiga sumber madu berbeda terhadap persentase ikan jantan dan ekspresi gen aromatase. Pada percobaan satu, 30 larva ikan nila berumur 12 hari setelah menetas direndam menggunakan madu hutan, madu ternak dan madu bakau, dengan dosis 10 ml L-1 air selama 10 jam. Ikan dipelihara dalam kondisi yang sama selama dua bulan. Hasil penelitian menunjukkan bahwa persentase ikan jantan tidak berbeda nyata antar perlakuan madu (p>0,05), tetapi semuanya berbeda nyata dengan kontrol (p<0,05). Pada percobaan kedua, larva ikan nila direndam dalam air mengandung dua bahan bioaktif madu, yakni chrysin dan kalium dengan dosis masing-masing 20 mg L-1 dan 0,026 g L-1. Ekspresi gen aromatase tipe gonad (aroma-g) dan tipe otak (aroma-o) dianalisis menggunakan metode RT-PCR. Sampel jaringan diambil pada waktu 1, 6, 12, 24, dan 48 jam pascaperlakuan madu, chrysin, dan kalium, serta setelah ikan berumur dua bulan.Ukuran fragmen DNA aromatase pada gonad betina sekitar 200 bp. Perendaman chrysin dan kalium meningkatkan persentase ikan jantan (p<0,1). Analisis RT-PCR menunjukkan bahwa madu, chrysin, dan kalium dapat menekan ekspresi gen aroma-g pada jam ke-12 pascaperendaman. Dengan demikian dapat disimpulkan bahwa madu, chrysin dan kalium dapat digunakan untuk pengarahan diferensiasi ikan nila, dan mekanis-menya seperti penghambat aromatase.

Promoter in the expression vector plays an important role on regulating of gene expression in transgenic fish. In fish transgenesis, researcher convinced that the use of all-fish expression vector is safety and prospective. The study was performed to isolate P-actin promoter, the promoter which has ubiquitous, constitutive, housekeeping characteristics, of common carp as a first step to construct all-common carp expression vector. Common carp P-actin promoter (ccBA) was isolated using PCR method with FBP1, RBP1, and RBP2 primers. Sequencing was performed using ABI PRISM 3100 machine, and analysis of sequences was conducted using GENETYX version 7 software. The results of sequence analysis showed that the length of DNA fragment obtained was approximately 1.5 kb. Results of homology with P-actin promoter sequence of a gene bank database (Accession No.: M24113) was 97.5%. The evolutionary conserved of transcription factor for P-actin promoter including CCAT, CArG, and TATA boxes were found in the sequence. Ubiquitous and higher expression of green fluorescent protein driven by ccBA promoter in muscle of common carp larvae was detected. It is most likely that the isolated sequence is a common carp P-actin promoter. AbstrakPromoter dalam vektor ekspresi berperan penting dalam mengatur ekspresi gen pada ikan transgenik. Dalam transgenesis ikan, peneliti yakin bahwa penggunaan vektor ekspresi semua ikan aman dan prospektif. Penelitian ini dilakukan untuk mengisolasi promoter P-aktin, promoter yang memiliki karakteristik ubiquitous, constitutive, house keeping, dari ikan mas sebagai langkah awal untuk mengkonstruksi vektor ekspresi semua ikan mas. Promoter P-aktin ikan mas (ccBA) diisolasi menggunakan metode PCR dengan primer FBP1, RBP1, dan RBP2. Sequensing dilakukan dengan menggunakan mesin ABI PRISM 3100, dan analisis sekuen dilakukan dengan menggunakan perangkat lunak GENE-TYX versi 7. Hasil analisis sekuen menunjukkan bahwa panjang fragmen DNA yang diperoleh adalah sekitar 1,5 kb. Hasil homologi dengan sekuen promoter P-aktin dari pangkalan data bank gen (No. Aksesi: M24113) adalah sebesar 97,5%. Faktor transkripsi yang tetap secara evolusioner untuk promoter P-aktin promoter termasuk CCAT, CArG, dan boks TATA ditemukan dalam sekuen. Ubiquitous dan ekspresi tertinggi protein pendaran hijau (GFP) dikendalikan oleh promoter ccBA dalam otot larva ikan mas yang dideteksi. Dengan demikian, kemungkinan besar bahwa sekuen yang terisolasi adalah promoter P-aktin ikan mas.

The purpose of this study was to describe the effect of dietary protein levels enriched with rGH on the ratio of length of the intestine to body weight of giant gourami. The diet with different protein levels (i.e. 21%, 28%, 34%; isoenergy) coated with egg yolk containing rGH. Each treatment has a control without rGH addition. Juveniles of giant gourami (body weight15.83 ± 0.13 g) were fed with a diet containing rGH twice a week. Fish were reared in 100 L glass aquaria with density of 10 fish per aquaria for 42 days. At the end of the treatment, body weight and length of the intestine were measured to determine the effect of treatment on the ratio of these two parameters. The result of research showed that at each level of treatment, the dietary protein levels and rGH supplementation gave a significant effect to increase the ratio of intestinal length to body weight (p <0,05). Ratio of intestinal length to body weight of fish was increase by decre-asing protein level in dietary (p <0,05). No interaction between protein levels and rGH additon on the ratio of intestinal length to body weight (p> 0,05). AbstrakPenelitian ini bertujuan untuk menjelaskan pengaruh pemberian pakan dengan kadar protein berbeda yang diperkaya dengan rGH pada nisbah panjang usus dan bobot tubuh ikan gurami. Pakan dengan kadar protein berbeda (21%, 28%, 34%;isoenergi) dibalut kuning telur yang mengandung rGH. Masing-masing perlakuan memiliki kontrol tanpa penam-bahan rGH. Yuwana ikan gurami (bobot tubuh15,83±0,13 g) diberi pakan mengandung rGH dua kali seminggu. Yuwa-na ikan dipelihara dalam akuarium volume100 L dengan padat tebar 10 ekor per akuarium selama 42 hari. Pada akhir perlakuan dilakukan pengukuran bobot tubuh dan panjang usus. Hasil penelitian menunjukkan bahwa pada masing-ma-sing taraf perlakuan kadar protein pakan, pemberian rGH berpengaruh nyata (p<0,05) terhadap peningkatan nisbah pan-jang usus dan bobot tubuh ikan. Penurunan kadar protein pada pakan terbukti meningkatkan nisbah panjang usus dan bobot tubuh ikan (p<0,05). Tidak terdapat interaksi antar perlakuan kadar protein dan penambahan rGH terhadap rasio panjang usus terhadap bobot tubuh (p>0,05).

The objective of this study was to compare the growth response of 3 different sizes of humpback grouper juveniles, which were come from the same brood stock group and spawning season after “stop and go” experiment of recombinant Epinephelus lanceolatus growth hormone (rElGH); that are with, without and retreated with rElGH for 42 days respectively. Each size group was divided into two treatment groups. The first group was treated with 50 mg crude rElGH kg-1 in commercial diet (pC) and a second group as a control. Weight gain of pC compares to control for small size group, medium size group and large size groups of juveniles subsequently for first stage were 85.89%, 39.66% and 16.34%; second stage were -34.57%, -14.76%, and -5.27%; and third stage were 56.16%, 50.24% and 59.14%. Specific growth rate differences of small, medium and large size of pC compared to control in first stage were 41.6%, 19.06% and 7.52%; second stage were -44.81%, -27.23% and -14.66%; and third stage were 55.9%, 40.62% and 48.42%. No significant difference of condition factor among all sizes of pC and control fish. Protein content and retention, and liver glycogen content from pooled sample of all size fish groups pC treatment in the second stage were decreasing compared to the first stage, respectively, 11.49%, 35.14% and 84.73%. It can be concluded that rElGH treatment improved growth performance of all size fish groups, however small juveniles have highest growth response compared to medium and large juvenile groups. The ceasing of rElGH treatment on second experiment stage is most likely causing the loss of accelerating growth factor then decreasing growth performance, protein content and retention, and liver glycogen content. AbstrakPenelitian bertujuan membandingkan respons pertumbuhan tiga kelompok ukuran benih ikan kerapu bebek dari kelompok induk dan periode pemijahan yang sama terhadap hormon pertumbuhan rekombinan ikan kerapu kertang Epine-phelus lanceolatus (rElGH); melalui eksperimen “putus dan sambung” yaitu dengan, tanpa, dan perlakuan kembali rElGH masing-masing selama 42 hari. Setiap kelompok ukuran dibagi menjadi dua kelompok perlakuan, kelompok pertama diberi perlakuan rElGH dengan dosis 50 mg rElGH-HP55 kg-1 pakan (pC) sedangkan kelompok kedua sebagai kontrol. Pertambahan bobot badan kelompok pC dibandingkan dengan kontrol pada benih berukuran kecil, sedang dan besar berturut-turut pada eksperimen tahap pertama 85,89%, 39,66% dan 16,34%; tahap kedua -34,57%, -14,76%, dan -5,27%, dan tahap ketiga 56,16%, 50,24% dan 59,14%. Perbedaan laju pertumbuhan spesifik benih berukuran kecil, se-dang dan besar perlakuan pC terhadap kontrol pada eksperimen tahap pertama 41,6%, 19,06% dan 7,52%; tahap kedua -44,81%, -27,23% dan -14,66%; dan tahap ketiga 55,9%, 40,62% dan 48,42%. Faktor kondisi pC dan kontrol pada se-mua kelompok ukuran tidak berbeda nyata. Kandungan dan retensi protein, dan kandungan glikogen hati gabungan sampel dari semua kelompok ukuran ikan perlakuan pC pada eksperimen tahap kedua menurun dibandingkan eksperi-men tahap pertama, masing-masing sebesar 11,49%, 35,14% dan 84,73%. Dapat disimpulkan pemberian rElGH mema-cu pertumbuhan semua kelompok ukuran benih ikan, namun benih berukuran kecil mempunyai respons pertumbuhan lebih tinggi daripada kelompok benih berukuran sedang dan besar. Penghentian pemberian rElGH menyebabkan ber-hentinya faktor pemacu pertumbuhan, sehingga performa pertumbuhan, kandungan dan retensi protein, dan kandungan glikogen hati menurun.

Bacterial disease in catfish has many detrimental to farmers, especially in the larvae stadium in the hatchery up to ready size of stocking seed. Therefore we need technology that can produce disease resistant of catfish. In that regard, one of the antimicrobial enzyme namely lysozyme which plays an important role in the maternal immunity can be introduced into the catfish genome. This study aimed to evaluate the success of the lysozyme gene insertion into the genome of striped catfish to generate maternal disease resistance. Gene transfer method was done by using electroporation on striped catfish spermatozoa. Lysozyme plasmid electroporation performed with a dose 100^g ml-1 and 125V.cm-1, pulse length 30 milliseconds with a pulse interval 0.1 milliseconds and pulse number 5 times. Electroporated spermatozoa were used to fertilize the egg. Lysozyme gene insertion success was indicated by tests performed on embryogenesis, larval, and seed stage. The test results at both DNA and RNA level showed a positive result. Individual that brings the lysozyme gene will be used as a candidate for breed line maternal immune as other ways to formation of resistant disease varieties. AbstrakSerangan penyakit bakterial pada ikan patin telah banyak merugikan para pembudi daya ikan patin terutama pada seg-men perbenihan hingga ukuran siap tebar. Oleh karena itu diperlukan teknologi yang mampu menghasilkan ikan patin yang tahan penyakit. Berkaitan dengan hal itu, salah satu enzim antimikroba yaitu lisozim yang memainkan peranan penting dalam imunitas bawaan dapat diintroduksikan ke dalam genom ikan. Penelitian ini bertujuan untuk mengevalu-asi keberhasilan insersi gen lisozim ke dalam genom ikan patin siam sebagai galur ikan patin tahan penyakit. Transfer gen dilakukan dengan menggunakan teknik elektroporasi pada spermatozoa ikan patin siam. Elektroporasi dilakukan dengan konstruksi gen lisozim berupa plasmid DNA dosis 100 ^g.ml-1 dengan voltase 125 V. cm-1, panjang kejutan 30 milidetik dengan interval kejutan 0.1 milidetik dan jumlah kejutan lima kali. Spermatozoa hasil elektroporasi digunakan untuk membuahi telur. Pengujian keberhasilan insersi gen lisozim dilakukan pada tahap embrio larva, dan pada benih. Hasil pengujian, baik pada tingkat DNA maupun pada tingkat RNA dari sampel spermatozoa dan larva (whole cell), memperlihatkan hasil yang positif. Individu ikan patin yang membawa gen lisozim dan telah terintegrasi ke dalam ge-nomnya akan digunakan sebagai kandidat dalam pembentukan galur ikan patin tahan penyakit.

Good sperm quality and inactive condition during the electroporation process determines the success of transgenesis. The study aims to observe the morphology and physiology of sperm, and evaluate the effects of different voltage and number of electric shocks on motility and viability of climbing perch sperm. Observation of morphological characters by microscope with a magnification of 400X, physiological character with various parameters, such as the level of os-molarity, volume, pH, density and total of sperm. The climbing perch sperm consisted of three parts, namely the ovoid head, middle, and tail. Sperm viability in the water was 116±34 seconds. Sperm quantity, volume of semen, pH, and sperm density were increased after ovaprim injection (p<0.05). Osmolarity levels of body and testes of climbing perch ranged at 390-405 mOsmol kg-1. Sperm diluted in 13 g L-1 NaCl solution could be kept for a maximum of 60 minutes at room temperature (26°C) and cold temperature (4°C), while without NaCl solution could be 360 minutes. Fertilization and hatching rates of eegs fertilized by sperm form the minced testes, and larval survival up to 10 days post-hatching were similar with semi-natural spawning. Electroporation of climbing perch sperm diluted in 13 g/L NaCl solution was successfully performed using a program with a long shock of 0.5 milliseconds and the interval shock of 0.1 seconds. Increasing the voltage and number of shock decreased motility and viability of sperm (p<0.05). Those electric shock parameters can potentially be used to generate climbing perch transgenic by electroporation method. AbstrakKualitas sperma yang baik dan dalam keadaan tak aktif selama proses elektroporasi sangat menentukan keberhasilan transgenesis. Penelitian ini bertujuan untuk mengeksplorasi karakter morfologis dan fisiologis sel sperma, serta menge-valuasi efek voltase dan jumlah kejut listrik berbeda terhadap motilitas dan viabilitas sperma ikan betok. Pengamatan karakter morfologi dilakukan pada mikroskop dengan pembesaran 400x, karakter fisiologi dengan berbagai parameter yakni tingkat osmolaritas, volume, pH, kepadatan dan total sperma. Hasil penelitian menunjukkan bahwa sperma ikan betok terdiri atas tiga bagian, yakni kepala yang berbentuk oval, bagian tengah, dan ekor sperma. Viabilitas sperma ikan betok dalam air adalah selama 116±34 detik. Induksi ovaprim dapat meningkatkan volume sperma, pH, kepadatan dan total sperma (p<0,05). Tingkat osmolaritas tubuh dan testis ikan betok (390-405 mOsmol kg-1). Penyimpanan sperma yang diencerkan dengan larutan NaCl 1,3% hanya dapat bertahan maksimum selama 60 menit pada suhu ruang (26°C) dan suhu dingin (4°C), sedangkan tanpa pengenceran dapat bertahan selama 360 menit. Derajat pembuahan dan kelangsungan hidup larva hingga 10 hari pascatetas menggunakan sperma dari testes cacah sama dengan pemijahan semi alami. Elektroporasi sperma ikan betok dengan pengencer larutan NaCl 1,3% berhasil dilakukan menggunakan lama kejut 0,5 milidetik, dan jeda antar kejut 0,1 detik. Peningkatan voltase dan jumlah kejut yang diberikan saling berinter-aksi menurunkan motilitas dan viabilitas sperma (p<0,05). Metode kejut listrik tersebut berpotensi digunakan untuk menghasilkan ikan betok transgenik dengan metode elektroporasi.

This study was aimed toenhance the growth of the giant gourami juve-nile by immersion with recombinant Cyprinus carpio growth hormone (rCcGH). Recombinant growth hormone was expressed by plasmid pCold-I/CcGHin E. coli BL21 (DE3). The inclusion bodies were isolated from E. coli using lyso-zyme and centrifugation method. Immersion with water containing 0.9% NaCl and 0.01% bovine serum albumin and inclusion bodies with different doses of rCcGH, 5 mgL-1 (C1), 15 mgL-1 (C3), and 30 mg L-1 (C6) was performed for 1 hour on weekly basis for the first four weeks of experimental period. Fish were subjected to salinity shock of 30 ppt NaCl for 2 minutes before it was transferred into the water containing rCcGH. Controls fish were without immersion and salinity shock (control), immersed in 30 ppt NaCl for 2 minutes (SS), immersed in 0.01% BSA (BSA), and immersed in medium containing BSA and 30 mg L-1 inclusion bodies of pCold without rCcGH (pCold).The giant gourami ju-venile treated with 30 mg L-1 rCcGH were 72.90% heavier and 21.04% longer (p<0.05) than the control, and 43.07% heavier and 14.64% longer (p<0.05) than the pCold. No significant difference in survival rate was obtained between treatments and controls. Biomass of fish treated 30 mgL-1 rCcGH was the highest among others. The rCcGH was able to promote soluble protein synthesis and lipid utilization as energy sources to spare protein (protein sparing effect). Thus, immersion with rCcGH could be applied to enhance the growth of giant gourami juvenile. AbstrakPenelitian ini bertujuan untuk memacu pertumbuhan benih ikan gurame melalui perendaman dengan hormon pertumbuhan rekombinan ikan mas (rCcGH). Hormon pertumbuhan rekombinan diekspresikan oleh plasmid pCold-I/CcGH di dalam Escherichia coli BL21(DE3). Badan inklusi diisolasi dari E. coli menggunakan lisozim dan metode sentrifugasi. Perendaman dilakukan selama 1 jam di dalam air yang mengandung 0,9% NaCl, 0,01% albumin serum sapi (BSA), dan badan inklusi rCcGH pada dosis5 mg L-1 (C1),15 mg L-1 (C3), dan 30mg L-1 (C6),sekali seminggu pada 4 minggu pertama. Ikan diberi kejutan salinitas 30 ppt NaCl selama dua menit sebelum ikan dipindahkan ke dalam air yang mengandung rCcGH.Sebagai kontrol ialah: benih ikan gurame tanpa perendaman (kontrol), benih ikan gurame yang diberi kejutan salinitas (SS), benih ikan gurame yang direndam di dalam air media yang mengandung BSA (BSA), dan benih ikan gurame yang direndam di dalam air yang mengandung BSA dan protein pCold-I tanpa rCcGH (pCold). Setelah tujuh minggu pemeliharaan, kelompok ikan yang direndam dalam air yang mengandung 30 mg L"1 rCcGH,72,90% bobot lebih berat dan 21,04% badan lebih panjang di-bandingkan dengan kontrol serta 43,07% bobot lebih berat dan 14,64% badan lebih panjang dibandingkan dengan pCold (p>0,05). Kelangsungan hidup antar perlakuan dan kontrol tidak berbeda nyata. Biomassa kelompok ikan yang direndam dengan 30 mg L-1 adalah yang tertinggi dibandingkan perlakuan lainnya. GH rekombinan ikan mas meningkatkan sintesis protein terlarut dan pemanfaatan lipid sebagai sumber energi sehingga mengoptimalkan pemanfaatan protein untuk pertumbuhan (protein sparing effect). Dengan demikian, perendaman dengan rCcGH dapat diaplikasikan untuk memacu pertumbuhan benih ikan gurame.