Akselerasi moulting larva udang Vaname (Litopenaeus vannamei) dengan pemberian kalsium hidroksida Ca(OH)2

Kualitas dan pertumbuhan larva udang vaname sangat dipengaruhi oleh proses dan frekuensi molting, dimana pertumbuhan erat kaitannya dengan proses pergantian cangkang. Kalsium hidroksida Ca(OH)2  merupakan salah satu mineral kalsium yang berhubungan dengan kadar kalsium kulit dan kadar kalsium lingkungan seiring pertukaran kalsium secara terus-menerus antara tubuh dan lingkungan. Tujuan penelitian ini adalah mengevaluasi akselerasi dan frekuensi molting larva udang vaname serta pengaruh lanjut Kalsium Hidroksida Ca(OH)2 terhadap laju pertumbuhan larva udang vaname. Desain penelitian adalah eksperimental menggunakan Rancangan Acak Lengkap dengan 4 taraf dosis Ca(OH)2 : A. Tanpa penambahan Ca(OH)2 (kontrol), B. 2 mg/ L air, C. 4 mg/ L air, dan D. 6 mg/L air. Hasil penelitian menunjukkan bahwa dosis Ca(OH)2 sebanyak 4 mg/L air menunjukkan intensitas/persentase molting sebesar 67%, laju pertumbuhan berat mutlak sebesar 2.35 mg, pertumbuhan panjang mutlak sebesar 4,93 mm, dan tingkat kelulushidupan udang (SR) sebesar 49%

Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii

Promoter regulates expression level of foreign gene in transgenic organism. This study was performed to select asuitable promoter as the fi rst step towards production of valuable trait-enhanced seaweed by transgenic technology. Greenfl uorescent protein (GFP) gene was used as a reporter to determine the activity of promoter in seaweed Kappaphycusalvarezii. GFP gene constructs driven by cytomegalovirus (pCMV-GFP), caulifl ower mosaic virus (pCaMV-GFP),medaka β-actin (pmBA-GFP) and Japanese fl ounder keratin (pJfKer-GFP) promoters were introduced by electroporationmethod. Electroporation was performed using a gene pulser (BIORAD) with voltage of 300 V, pulse length of 0.5 ms,pulse numbers of 4, and pulse interval of 0.1 s. Promoter activity was determined by analyzing GFP gene expressionlevel using a fl uorescent microscope. The results showed that CMV regulated highest number of fi lament callus(34.10%±1.49) expressing GFP at medium to strong fl uorescence levels. CaMV promoter had relatively similar activitywith CMV, but lower number of fi lament callus expressing GFP (10.48%±0.25). mBA promoter drove GFP expressionat medium level and similar number of fi lament callus (8.85%±2.31) expressing GFP with CaMV, while JfKer promoterhad lowest activity by means in number of fi lament callus expressing GFP (4.79%±0.26) and GFP expression level. PCRanalysis for transgenic confi rmation showed a DNA band of PCR product from pCMV-GFP and pCaMV-GFP expressingfi lament callus in the same size (about 0.6 kb) with positive control of plasmid. Thus, CMV and CaMV promoters wasan appropriate promoter and foreign gene could be transferred to fi lament callus by electroporation method. Combiningthis achievement with developing a culture method of fi lament callus to be thallus, stable transgenic breeding in K.alvarezii can be feasible

Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii

Promoter regulates expression level of foreign gene in transgenic organism. This study was performed to select a suitable promoter as the fi rst step towards production of valuable trait-enhanced seaweed by transgenic technology. Green fl uorescent protein (GFP) gene was used as a reporter to determine the activity of promoter in seaweed Kappaphycus alvarezii. GFP gene constructs driven by cytomegalovirus (pCMV-GFP), caulifl ower mosaic virus (pCaMV-GFP), medaka β-actin (pmBA-GFP) and Japanese fl ounder keratin (pJfKer-GFP) promoters were introduced by electroporation method. Electroporation was performed using a gene pulser (BIORAD) with voltage of 300 V, pulse length of 0.5 ms, pulse numbers of 4, and pulse interval of 0.1 s. Promoter activity was determined by analyzing GFP gene expression level using a fl uorescent microscope. The results showed that CMV regulated highest number of fi lament callus (34.10%±1.49) expressing GFP at medium to strong fl uorescence levels. CaMV promoter had relatively similar activity with CMV, but lower number of fi lament callus expressing GFP (10.48%±0.25). mBA promoter drove GFP expression at medium level and similar number of fi lament callus (8.85%±2.31) expressing GFP with CaMV, while JfKer promoter had lowest activity by means in number of fi lament callus expressing GFP (4.79%±0.26) and GFP expression level. PCR analysis for transgenic confi rmation showed a DNA band of PCR product from pCMV-GFP and pCaMV-GFP expressing fi lament callus in the same size (about 0.6 kb) with positive control of plasmid. Thus, CMV and CaMV promoters was an appropriate promoter and foreign gene could be transferred to fi lament callus by electroporation method. Combining this achievement with developing a culture method of fi lament callus to be thallus, stable transgenic breeding in K. alvarezii can be feasible

KAJIAN INDUKSI KALUS RUMPUT LAUT Kappaphycus alvarezii UNTUK PRODUKSI EMBRIO SOMATIK

Untuk mendukung program transgenesis pada rumput laut, embrio somatik dapat digunakan sebagai material untuk transfer gen baik secara individu sel ataupun kluster sel embriogenik, sehingga mempercepat keberhasilan dengan peluang transformasi yang lebih tinggi. Penelitian ini bertujuan untuk mengkaji induksi kalus rumput laut K. alvarezii untuk produksi sel embrio somatik (e.s.) dengan beberapa rasio zat pengatur tumbuh (ZPT) dan konsentrasi agar media induksi, sampai sel menjadi filamen. Penelitian terdiri atas dua tahap: Tahap (1) induksi kalus, dengan rasio ZPT asam indol asetat (IAA):kinetin = 0,5:0,0 mg/L; 1,0:1,0 mg/L; dan 2,0:0,2 mg/L dengan konsentrasi agar media induksi = 0,6%; 0,8%; 1,0%; dan 1,5%. Tahap (2) regenerasi massa sel e.s., dengan rasio IAA:kinetin = 0,1:1,0 mg/L; 0,0:0,1 mg/L dan tanpa ZPT dengan konsentrasi agar media = 0,4%; 0,6%; dan 0,8%. Untuk perkembangan sel-sel e.s. lebih lanjut dipelihara pada kultur cair. Hasil penelitian menunjukkan pada tahap induksi kalus, rasio IAA: kinetin = 1:1 mg/L dengan konsentrasi agar media 0,8% dan 1,0% menghasilkan persentase induksi kalus tertinggi (90%). Pada tahap regenerasi massa sel e.s., ZPT tidak berpengaruh terhadap perkembangan massa sel e.s., di mana tanpa ZPT dengan konsentrasi agar 0,6% memperlihatkan perkembangan tertinggi (rata-rata diameter massa sel 5 mm). Pada media cair, perkembangan sel e.s. dari single cell ukuran 3-4 mm menjadi filamen-filamen ukuran rata-rata 0,5 mm dapat dicapai dalam satu bulan kultur. Keberhasilan produksi sel e.s. K. alvarezii, selain sebagai material untuk transfer gen juga dapat dijadikan acuan dalam produksi benih rumput laut kultur jaringan.To support the program of seaweed transgenesis, somatic embryo can be used as a materials for gene transfer purpose either by individual or cluster of cells in accelerating the higher rate of transformation. This research aims to study the callus induction of seaweed K. alvarezii for production of somatic embrio (s.e) cell by different ratio of growth regulators (GR) and agar media concentrations. The study consists of two stages: stage (1) callus induction to the cells filament using GR of indol acetic acid (IAA):kinetin in ratio of  0.5:0.0 mg/L; 1.0:1.0 mg/L; and 2.0:0.2 mg/L on the media agar concentration of 0.6%; 0.8%; 1.0%; and 1.5%, and stage (2) regeneration of the cell mass of s.e, using GR of IAA:kinetin in ratio of 0.1:1.0 mg/L; 0.0:0.1 mg/L on the media agar concentration of 0.4%; 0.6%; and 0.8%. For further maintenance, the s.e cells were cultured in liquid media. The results of callus induction showed that the ratio of IAA:kinetin (1:1) on the media agar concentration of 0.8% and 1.0% produced the highest callus induction (90%). The study of mass cell regeneration of s.e showed did not different of GR IAA:kinetin ratio of the cell mass development, but the media agar concentration of 0.6% and 0.4% showed the higher growth of cell mass (in diameter size of 4-5 mm). The development of cells culture of s.e from the size of 3-4 mm to filament of 0.5 mm could be reached in one month of culture period. The success of K. alvarezii s.e production will be helpful not only as a material for gene transfer but also as a reference on tissue culture for seed production of seaweed

KAJIAN INDUKSI KALUS RUMPUT LAUT Kappaphycus alvarezii UNTUK PRODUKSI EMBRIO SOMATIK

Untuk mendukung program transgenesis pada rumput laut, embrio somatik dapat digunakan sebagai material untuk transfer gen baik secara individu sel ataupun kluster sel embriogenik, sehingga mempercepat keberhasilan dengan peluang transformasi yang lebih tinggi. Penelitian ini bertujuan untuk mengkaji induksi kalus rumput laut K. alvarezii untuk produksi sel embrio somatik (e.s.) dengan beberapa rasio zat pengatur tumbuh (ZPT) dan konsentrasi agar media induksi, sampai sel menjadi filamen. Penelitian terdiri atas dua tahap: Tahap (1) induksi kalus, dengan rasio ZPT asam indol asetat (IAA):kinetin = 0,5:0,0 mg/L; 1,0:1,0 mg/L; dan 2,0:0,2 mg/L dengan konsentrasi agar media induksi = 0,6%; 0,8%; 1,0%; dan 1,5%. Tahap (2) regenerasi massa sel e.s., dengan rasio IAA:kinetin = 0,1:1,0 mg/L; 0,0:0,1 mg/L dan tanpa ZPT dengan konsentrasi agar media = 0,4%; 0,6%; dan 0,8%. Untuk perkembangan sel-sel e.s. lebih lanjut dipelihara pada kultur cair. Hasil penelitian menunjukkan pada tahap induksi kalus, rasio IAA: kinetin = 1:1 mg/L dengan konsentrasi agar media 0,8% dan 1,0% menghasilkan persentase induksi kalus tertinggi (90%). Pada tahap regenerasi massa sel e.s., ZPT tidak berpengaruh terhadap perkembangan massa sel e.s., di mana tanpa ZPT dengan konsentrasi agar 0,6% memperlihatkan perkembangan tertinggi (rata-rata diameter massa sel 5 mm). Pada media cair, perkembangan sel e.s. dari single cell ukuran 3-4 mm menjadi filamen-filamen ukuran rata-rata 0,5 mm dapat dicapai dalam satu bulan kultur. Keberhasilan produksi sel e.s. K. alvarezii, selain sebagai material untuk transfer gen juga dapat dijadikan acuan dalam produksi benih rumput laut kultur jaringan. To support the program of seaweed transgenesis, somatic embryo can be used as a materials for gene transfer purpose either by individual or cluster of cells in accelerating the higher rate of transformation. This research aims to study the callus induction of seaweed K. alvarezii for production of somatic embrio (s.e) cell by different ratio of growth regulators (GR) and agar media concentrations. The study consists of two stages: stage (1) callus induction to the cells filament using GR of indol acetic acid (IAA):kinetin in ratio of  0.5:0.0 mg/L; 1.0:1.0 mg/L; and 2.0:0.2 mg/L on the media agar concentration of 0.6%; 0.8%; 1.0%; and 1.5%, and stage (2) regeneration of the cell mass of s.e, using GR of IAA:kinetin in ratio of 0.1:1.0 mg/L; 0.0:0.1 mg/L on the media agar concentration of 0.4%; 0.6%; and 0.8%. For further maintenance, the s.e cells were cultured in liquid media. The results of callus induction showed that the ratio of IAA:kinetin (1:1) on the media agar concentration of 0.8% and 1.0% produced the highest callus induction (90%). The study of mass cell regeneration of s.e showed did not different of GR IAA:kinetin ratio of the cell mass development, but the media agar concentration of 0.6% and 0.4% showed the higher growth of cell mass (in diameter size of 4-5 mm). The development of cells culture of s.e from the size of 3-4 mm to filament of 0.5 mm could be reached in one month of culture period. The success of K. alvarezii s.e production will be helpful not only as a material for gene transfer but also as a reference on tissue culture for seed production of seaweed

Activity of Natural Compound Pothos tener Wall on Aeromonas hydrophila Infection to Prevent of Antibiotics

This study aims to discover how influential Photos tener Wall is as an anti-bacterial treatment for Aeromonas hydrophila. Methods: Cyprinus Carpio were reared for 28 days, and on the 29th and 30th days before Aeromonas hydrophila infection, the fish were adequately fasted. On the 31st day, they were intramuscularly challenged with A. hydrophila (105 CFU/mL) (the first day in A. hydrophila infection). The treatments given were (I) immersed trial with fresh (live) P tener Wall: (H1) Immersed with 15 g of P. tener Wall plant, (H2) Immersed with 30 g of P. tener Wall plant, and (H3) Immersed with 60 g of P. tener; (II) feeding trials in which the treatments given were (P1) 2% of P. tener Wall powder mixed with 1 kg commercial diet and (P2) 4% of P. tener Wall powder mixed with 1 kg commercial diet; Experiment III combined the best results from experiment I (H2) and experiment II (P2) and Oxytetracycline 5 g/kg feed as a control antibiotic. The result obtained was that the treatment of 30 g of fresh P. tener Wall or adding 4% simplicial P. tener Wall in the diet could increase koi fish's immune response and resistance to A. Hydrophila has a survival rate that reaches 100%. This treatment has the same effect as using antibiotic Oxytetracycline 5 g/kg of feed. An important aspect for further research is that P. tener wall can be tested on other fish diseases caused by bacteria or fungi