Clarias gariepinus growth hormone (CgGH) transgenesis was previously used to develop a population of second-generation (G2) transgenic mutiara catfish (C. gariepinus). The third generation of these fish (1-month old fingerlings) had 2–3 times improvement in growth compared to non-transgenic fish in the commercial feed test for 6 weeks of rearing. We assessed the impact of CgGH transgene expression on growth and protein and amino acid content of the G3 generation of these transgenic mutiara catfish relative to non-transgenic catfish. Since variation in composition of feed mixes can affect protein and amino acid content of fish, we tested three mixtures of commercial feed and boiled tuna (Euthynnus affinis): feed A (50:50 feed to tuna), B (65:35), and C (80:20) to transgenic catfish. Feed A* (50:50) was fed as a control to non-transgenic catfish. Feed efficiency, including feed conversion ratio and protein use efficiency (i.e., protein retention and protein productive value), was assessed. Feed efficiency, protein content, and essential amino acid content in G3 transgenic catfish (feed A and B) were higher than in non-transgenic fish (feed A*). The latter were deficient in lysine and methionine. Transgenic catfish fed with feed C (80:20) showed lysine deficiency and lower growth than fish fed feeds A and B. Feed B (65:35) was the optimal feed mixture utilized; it increased growth, protein levels, and feed conversion efficiency in G3 catfish. The growth of transgenic fish was higher than non-transgenic fish when supported by feeding with balanced nutrients.
This study aims to reveal effective concentration of moringa leaf extract which is inducing body resistance of sangkuriang catfish against Aeromonas hydrophila. This study used as experimental method of Completely Randomized Design (CRD) with six treatments and three replications. The treatment used in this research are the addition of Moringa leaf extract into feed with concentration of 0 ppm (A), 100 ppm (B), 125 ppm (C), 150 ppm (D), 175 ppm (E) and 200 ppm (F). Observed Parameters are total leukocyte count, increase in leukocyte, clinical sign and survival rate. Increase in leukocyte and survival rate data were analyzed using F test and Duncan test with significance level 0,05, while clinical sign data was analyzed descriptively. The results showed that Moringa leaf extract was effective to induce the immune of sangkuriang catfish. The 150 ppm concentration resulted the highest total leukocyte count of 23.46 ± 6.46% and didn’t have clinical symptons after challenge with Aeromonas hydrophila and highest survival rate 80.0 ± 5.00%. Based on the regression analysis, 153.95 ppm of Moringa leaf extract is the optimum concentration with 95.62% on the survival of sangkuriang catfish after challenge with Aeromonas hydrophila.AbstrakPenelitian ini bertujuan untuk menentukan konsentrasi yang efektif dari ekstrak daun kelor untuk menginduksi ketahanan tubuh ikan lele sangkuriang terhadap serangan bakteri Aeromonas hydrophila. Penelitian ini menggunakan metode eksperimental Rancangan Acak Lengkap dengan enam perlakuan dan tiga ulangan. Perlakuan penelitian yang digunakan adalah penambahan ekstrak daun kelor ke dalam pakan dengan konsentrasi 0 ppm (A), 100 ppm (B), 125 ppm (C), 150 ppm (D), 175 ppm (E) dan 200 ppm (F). Parameter yang diamati meliputi jumlah total sel darah, peningkatan jumlah sel darah putih, gejala klinis, dan sintasan. Data peningkatan jumlah sel darah putih dan sintasan dianalisis menggunakan uji F dan uji jarak berganda pada taraf kepercayaan 5%, sedangkan data gejala klinis dianalisis secara deskriptif. Hasil penelitian menunjukkan ekstrak daun kelor efektif untuk menginduksi ketahanan tubuh benih lele sangkuriang. Konsentrasi 150 ppm menghasilkan rata-rata jumlah sel darah putih tertinggi 23,46±6,46% dan setelah diuji tantang tidak menunjukkan gejala klinis, serta tingkat sintasan yang tertinggi sebesar 80,0±5,00%. Berdasarkan analisis regresi konsentrasi ekstrak daun kelor memberikan pengaruh sebesar 95,62% dengan konsentrasi optimum sebesar 153,95 ppm terhadap sintasan ikan lele sangkuriang setelah di uji tantang dengan Aeromonas hydrophila.
Aplikasi teknologi transfer gen melalui perantaraan sperma dimungkinkan untuk memperbaiki kualitas genetik ikan. Keberhasilan transfer gen hormon pertumbuhan (Growth Hormone =GH) ikan budidaya melalui elektroporasi sperma dapat menghasilkan ikan transgenik. Pada penelitian ini dilakukan transfer konstruksi vektor ekspresi gen GH lele dumbo (pTarget-CMV-CgGH) ke sperma lele dumbo (strain mutiara) menggunakan kejut listrik 125 Vcm-1 dengan jumlah pulsa 3 dan 5 serta 50 Vcm-1 dengan jumlah pulsa 3 dan 5. Konsentrasi vektor yang digunakan 60 ng/?L, lama kejutan 30 milidetik dan interval kejut 0,1 detik. Dari hasil elektroporasi sperma lele mutiara sebesar 125 Vcm-1 (jumlah pulsa 3 dan 5) serta 50 Vcm-1 (jumlah pulsa 3 dan 5) ini, terdeteksi fragmen berukuran 600 bp (gen GH lele dumbo) pada sperma dan fragmen 1000 bp (96 % identik dengan GH Clarias gariepinus, no. aksesi AF416488.1) pada ikan umur 4 bulan menunjukkan keberhasilan transfer gen GH lele dumbo (GH eksogen) ke sperma lele mutiara. Rata-rata kelipatan tumbuh ikan lele mutiara transgenik umur 4 bulan (125 Vcm-1 dan 50 Vcm-1 dengan jumlah pulsa 3 dan 5) berturut- turut 3,53 kali; 3,05 kali; 2,75 kali dan 2,76 kali dari ukuran ikan non transgenik (over-ekspresi GH eksogen). GH eksogen terdeteksi pada ikan lele mutiara transgenik dan tidak terdeteksi pada ikan lele mutiara non transgenik (non elektroporasi). Ikan lele mutiara transgenik berhasildirakit dengan menyisipkan gen GH lele dumbo menggunakan teknik elektroporasi sperma.
Isolasi gen hormon pertumbuhan lele dumbo (C. gariepinus) dari hipofisa ikanmerupakan sumber DNA yang selanjutnya dapat digunakan sebagai cetakan untuk mengkopi gen tersebut menggunakan primer Amc-GH-F dan Amc-GH-R. Fragmen DNA lele dumbo hasil isolasi terdeteksi berdasar elektroforesis gel agarosa 1%. Amplifikasi gen penyandi hormon pertumbuhan (Growth Hormone / GH) lele dumbo menggunakan primer Amc-GH-F (5’-GCAGAAATGGCTCGAGGTAAGG-3’) dan Amc-GH-R (5’-CAGGGTGCAGTTGGAATCC-3’) dapat mengkopi sekuen gen GH lele dengan ukuran fragmen PCR sekitar 1.400 bp. Sementara amplikon gen GH American catfish (Rhamdia quelen) menggunakan primer Amc-GH-F dan Amc-GH-R sebesar 1.465 bp. Hasil analisis sekuensing gen penyandi GH menggunakan program BlastP dan Genetyx versi 7.0, menunjukkan bahwa sekuen gen penyandi GH lele dumbo memiliki homologi 80% dengan sekuen GH C. gariepinus pada bank gen (no. aksesi AF 416488.1), sehingga sebagian besar sekuen gen penyandi hormon pertumbuhan ikan tersebut dapat diamplifikasi secara in vitro.
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