Identification of growth hormone (GH) gene in a target fish is the first step in the construction of “all fish genes transfer vector” to generate transgenic fish. The research was done to identify and characterize the GH gene of Pangasionodon hypophthalmus. There were several activities performed in identifying the GH gene: RNA extraction, cDNA synthesis, PCR amplification, and DNA fragment isolation. The characterizations were done using the nucleotide sequencing engine ABIPRISM 3100. The results were then analyzed using BLASTN/P and GENETYX version 7 program. The full-length GH gene of P. hypophthalmus was 1151 bp in length, coding for an open reading frame (ORF) of 603 bp. The 5’ and 3’ untranslated regions of the GH gene were 22 bp and 526 bp long, respectively. The GH gene of P. hypophthalmus had some common characteristics that are owned by GH genes, such as single tryptophan residue (W) on the 104th amino acid, 5 cysteine residues (C) on the amino acid 71, 135, 173, 190, and 198 and a motif of Asn-Xaa-Thr on C terminus which is the potential location for N-linked glycosilation. Polyadenylation signal (aataaa) was on the 14 bp at the upstream of polyadenylation location. Growth hormone of P. hypophthalmus consisted of over 200 amino acids from GH cDNA deduction. The highest proportion of amino acid composition was leusin (14%) while the lowest was tryptophan (0.5%).
The success of transgenic fish production has been achieved through eggs fertilization using electroporated sperms carrying exogenous DNA. This study was conducted in order to obtain the optimal electroporation condition for stripped catfish sperm. A plasmid containing green fluorescent protein (GFP) gene driven by carp β-actin promoter was transferred into sperm using electrophoresis method towards transgenic stripped catfish (Pangasionodon hypophthalmus) production. Electroporation was carried out using square wave shock with pulse length of 30 ms and pulse interval of 0.1 sec. Treatments are combination between voltage (50 V, 75 V, and 100 V) and pulse number (1 and 3). Exogenous DNA concentration used was 10 μg/mL of Tris-EDTA. Results showed that increasing the voltage from 50 to 100 decreased sperm motility, while pulse number did not affect sperm motility. Voltage of 50 gave the best motility of sperm, although sperm viability relatively similar between treatments and control except at 100 V with 3 pulses number. Further, electroporation-treated sperms were able to fertilize eggs. Higher hatching rate of eggs was obtained in electroporation treatment at 50 V with pulse number of 1 and 3. The persistence of transferred GFP was detected in electroporated and incubated sperms (control). However, GFP was only detected in larvae from eggs that were fertilized by electroporated sperm. Thus, electroporation could be applied to produce transgenic stripped catfish.
Penggunaan konsentrasi DNA yang tinggi dalam elektroporasi sperma meningkatkan pengikatan DNA eksogen pada sperma, dan meningkatkan persentase ikan yang membawa gen asing. Pada penelitian ini, konstruksi gen pCcBA-PhGH yang mengandung promoter β-aktin ikan mas (pCcBA) dan cDNA hormon pertumbuhan (PhGH) ikan patin siam (Pangasianodon hypophthalmus) dibuat dan selanjutnya ditransfer menggunakan metode elektroporasi pada sperma yang berperan sebagai perantara. Elektroporasi dilakukan dengan tipe kejutan square wave dengan panjang kejutan (pulse length) 30 milidetik, interval kejutan (pulse interval) 0,1 detik, kuat medan listrik (electric field strength) 125 V/cm, dan jumlah kejutan (pulse number) 3 kali. Hasil penelitian menunjukkan bahwa keberhasilan transfer gen PhGH eksogen pada ikan patin siam meningkat dengan meningkatnya konsentrasi DNA yang digunakan. Persentase ikan yang membawa gen asing pada konsentrasi DNA 10 μg/mL, 50 μg/mL, dan 90 μg/mL, secara berturut-turut adalah 28,57%; 78,57%; dan 85,71%. Bobot ratarata yuwana ikan patin siam transgenik F0 umur 2 bulan yang dihasilkan menggunakan konsentrasi DNA 50 μg/mL dan 90 μg/mL adalah 22,6% dan 19,0% lebih berat dibandingkan non-transgenik, tetapi pada konsentrasi 10 μg/mL lebih rendah (-8.45%). Populasi yuwana ikan patin siam berumur 4 bulan yang diintroduksi gen asing dengan konsentrasi 90 μg/mL memiliki bobot rataan 53,38% lebih berat dibandingkan kontrol non-transgenik. Dengan menggunakan metode RT-PCR, ekspresi gen PhGH terdeteksi pada sirip ikan transgenik, sedangkan pada ikan non-transgenik tidak terdeteksi. Dengandemikian, elektroporasi sperma menggunakan konsentrasi DNA 90 μg/mL efektif meningkatkan keberhasilan transfer gen, dan over-ekspresi gen PhGH eksogen meningkatkan pertumbuhan ikan patin siam.
Ikan lele Afrika (Clarias gariepinus) merupakan spesies asli Afrika yang telah diintroduksikan dan dibudidayakan secara komersial di Indonesia. Upaya peningkatan efisiensi produksi ikan lele terus ditingkatkan guna meningkatkan keuntungan. Salah satu upaya untuk meningkatkan efektivitas budidaya ikan lele adalah melalui penggunaan probiotik. Penelitian ini bertujuan untuk mengevaluasi pemanfaatan probiotik pada pembesaran ikan lele. Perlakuan yang diberikan berupa pemberian pakan hasil fermentasi probiotik dan pakan tanpa fermentasi (kontrol) dengan tiga ulangan. Pengujian dilakukan pada kolam terpal berukuran 3 m3 dengan padat tebar yang digunakan yaitu 500 ekor/wadah dan dipelihara selama 35 hari. Berdasarkan hasil pengujian, pemberian probiotik pada pakan dengan cara fermentasi mampu meningkatkan bobot dan biomassa panen secara signifikan (P<0,1). Bobot akhir ikan lele yang diberi pakan hasil fermentasi probiotik mencapai 76,9 ± 0,2 g; sedangkan kontrol 74,2 ± 0,2 g. Biomassa akhir ikan lele yang diberi pakan hasil fermentasi probiotik mencapai 37,91 ± 0,29 kg; sedangkan kontrol 34,65 ± 1,70 kg. Pemberian pakan yang difermentasi probiotik mampu meningkatkan retensi protein sebesar 1,02%; retensi karbohidrat sebesar 10,26%; dan retensi lemak sebesar 7,22%. Selain itu, penggunaan probiotik mampu menekan biaya produksi sebesar Rp 561,00/kg dan meningkatkan keuntungan sebesar 5%.African catfish (Clarias gariepinus) is a native African species that has been introduced and cultivated commercially in Indonesia. Efforts to increase the efficiency of catfish production were conducted in order to increase profit. One effort to increase the effectiveness of catfish farming is through the use of probiotics. This study was aimed to evaluate the use of probiotics in catfish farming. The treatments were fermented feed by probiotic and non fermented feed (control) and repeated three times. The experiment was conducted on a 3 m3 tarpaulin pond, with a density 500 fishes/pond, and reared for 35 days. Based on the results, the fermented feed by probiotic could increase the weight and biomass of harvested fish significantly. The weight of catfish fed with fermented feed reached 76.9 ± 0.2 g while the control was 74.2 ± 0.2 g. The final biomass of catfish fed with fermented feed reached 37.91 ± 0.29 kg while the control was 34.65 ± 1.70 kg. Feeding fermented feed by probiotic Gut Bio Aero increased the protein retention by 1.02%, carbohydrate retention by 10.26%, and lipid retention by 7.22%. The application of probiotic could reduce production cost by 561 IDR/kg and increased profit by 5%.
ABSTRAKSalah satu jenis ikan nila yang memiliki potensi nilai ekonomis yang tinggi dan dapat dibudidayakan di air tawar maupun di air payau adalah ikan nila merah. Dalam rangka pengembangannya diperlukan upaya untuk meningkatkan kualitas genetik di antaranya dengan cara seleksi. Kegiatan ini bertujuan untuk mengetahui estimasi nilai heritabilitas dan respons seleksi populasi jantan dan betina F-0 ikan nila merah hasil seleksi. Pemijahan dilakukan secara full-sib dengan perbandingan jantan dan betina 1:1 dan menghasilkan 10 famili dari 50 famili yang dibentuk. Pembesaran benih dilakukan di tambak bersalinitas 30 ppt menggunakan waring berukuran 3 m x 5 m x 1,5 m dengan padat tebar 10 ekor/m 2 . Seleksi calon induk pada populasi jantan dan betina ikan nila merah dilakukan setelah mencapai bobot 200-300 g. Cut off seleksi ditentukan pada bobot berkisar 225-354 g (jantan) dan 201-258 g (betina). Parameter yang diamati meliputi koefisien keragaman panjang dan bobot, diferensial seleksi, estimasi nilai heritabilitas, dan respons seleksi. Hasil penelitian menunjukkan koefisien keragaman karakter panjang pada populasi jantan berkisar dari 7,89%-11,18% dan bobot 21,22%-34,12% dengan diferensial seleksi 45,87-115,18 g; sedangkan populasi betina sebesar 6,08%-13,22% dan 16,39%-31,55%; serta 46,12-71,67 g. Hasil analisis estimasi nilai heritabilitas (dalam arti luas) pada karakter bobot ikan nila merah jantan 0,47±0,19 dan betina 0,19±0,11. Adapun prediksi respons seleksi yang akan diperoleh adalah 33,06 g (jantan) dan 11,65 g (betina). KATA KUNCI: heritabilitas, respons seleksi, ikan nila merah, payau ABSTRACT:Heritability estimates and response to selection of red tilapia in brackishwater pond.
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