, Alimuddin
Department of Aquaculture

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Growth and survival of post-larval Pacific white shrimp administered recombinant fish growth hormone by different immersion time

Jurnal Akuakultur Indonesia Vol 12, No 2 (2013): Jurnal Akuakultur Indonesia
Publisher : Jurnal Akuakultur Indonesia

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Abstract

This research was conducted to determine the optimum immersion time of recombinant giant grouper growthhormone (Ephinephelus lanceolatus; rElGH) at a dose of 15 mg/L that generated highest growth of white shrimppost-larvae (PL). The treatments were immersion time for one, two, and three hours. Two types of control wasprovided, namely it was without any treatment (control), and immersion in water containing 0.01% bovine serumalbumin (BSA) and total protein of Escherichia coli without rElGH (pCold control). All treatments and controlswere consisted of three replications. A total of 1,500 PL-2 shrimp were bath-immersed in a plastic packingcontaining 1 L of sea water, 15 mg/L rElGH, and 0.01% BSA. PL was further reared for 21 days in the 60 L glassaquarium, and fed nauplii Artemia two times and flake commercial diet five times daily, at satiation. The resultsshowed that the highest of biomass (36.29±1.46 g), specific growth rate (29.81±0.87%/day), and body length(20.08±0.42 mm) were obtained in three hours immersion treatment (p<0.05). Biomass of PL in three hoursimmersion treatment was approximately 66.0% higher compared to the control (21.87±2.53 g). Survival of shrimpin all treatment and control were similar (p>0.05). Thus, growth of white shrimp PL could be improved by bathimmersion for three hours in rElGH solution of 15 mg/L water.Keywords: recombinant growth hormone, different immersion time, Pacific white shrimp, biomass   Abstrak Penelitian ini dilakukan untuk menentukan lama waktu perendaman hormon pertumbuhan rekombinan ikankerapu kertang (Ephinephelus lanceolatus; rElGH) dosis 15 mg/L yang menghasilkan pertumbuhan tertinggi padapascalarva (PL) udang vaname. Perlakuan yang diberikan adalah lama perendaman PL-2, yaitu satu, dua, dan tigajam. Dua jenis kontrol dibuat yakni udang vaname PL-2 tidak diberi perlakuan (kontrol), dan direndam dalam airmengandung serum albumin sapi (BSA) 0,01% dan protein Escherichia coli tanpa rElGH (kontrol pCold). Setiapperlakuan dan kontrol diberi tiga ulangan. Sebanyak 1.500 ekor PL-2 direndam dalam kantong plastik kemasanberisi 1 L air laut mengandung rElGH 15 mg/L, dan BSA 0,01%. Selanjutnya, udang dipelihara selama 21 hari didalam akuarium volume 60 L, dan diberi pakan naupli Artemia sebanyak dua kali dan pakan komersial berbentukflake sebanyak lima kali sehari hingga kenyang. Hasil penelitian menunjukkan bahwa biomassa (36,29±1,46 g),pertumbuhan bobot spesifik (29,81±0,87%/hari), dan panjang tubuh (20,08±0,42 mm) tertinggi (p<0,05) diperolehpada perlakuan perendaman selama tiga jam. Biomassa udang perlakuan perendaman selama tiga jam lebih tinggi66% dibandingkan dengan kontrol (21,87±2,53 g). Kelangsungan hidup udang yang direndam dengan rElGH,kontrol dan kontrol pCold tidak berbeda nyata (p<0,05). Dengan demikian, pertumbuhan PL udang vaname dapatditingkatkan melalui perendaman selama tiga jam dalam larutan rElGH 15 mg/L air.Kata kunci: hormon pertumbuhan rekombinan, lama perendaman, pascalarva udang vaname, biomassa

Testicular cell transplantation of neon tetra Paracheirodon innesi into common carp fry

Jurnal Akuakultur Indonesia Vol 12, No 2 (2013): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT Neon tetra Paracheirodon innesi is an ornamental fish that have high export value. However, production is still relatively low due to low fecundity (approximately 180 eggs/female). Technology of testicular cell transplantation of neon tetra as donor to common carp as recipient fish which have high fecundity provides a promising way to overcome problem of neon tetra production. This research was performed to determine the optimum age of common carp fry that is able to receive donor cells and allow high success of transplantation. In this research, the testes of neon tetra fish were dissociated by 0.5% trypsin solution. The testicular cells were labeled with PKH-26 fluorescent dye, and then transplanted into the peritoneal cavity of seven, ten, and 14 days post hatching common carp fry. The results showed that the survival of seven day-old transplanted fry (31.25%) was lower than that of ten day-old (37.75%) and 14 day-old transplanted fry (56.25%). Percentage of fish colonized testicular cells donor at 21 days post-transplantation on seven days old and ten days old fry were similar (80%), while on 14 day-old fry was 60%. Based on the cumulative transplantation success rate (survival and colonization rates), transplantation on 14 days old fry (33.75%) showed higher result compared to transplantation on seven days old fry (25.00%) and ten day-old fry (30.00%). It can be concluded that transplantation of neon tetra testicular cells to common carp fry have been successfully carried out, and the optimum age of common carp fry to transplantation was 14 days after hatching. Keywords: transplantation, colonization, testicular cells, common carp, neon tetra  ABSTRAK Ikan neon tetra Paracheirodon innesi merupakan ikan hias yang memiliki nilai ekspor yang tinggi. Namun demikian, tingkat produksinya masih relatif rendah karena fekunditas ikan neon tetra yang sedikit (sekitar 180 telur/induk). Teknologi transplantasi sel testikular ikan neon tetra (ikan donor) ke ikan mas yang memiliki fekunditas telur yang banyak dan diharapkan mampu mengatasi ketersediaan benih ikan neon tetra. Penelitian ini bertujuan untuk menentukan umur optimum benih ikan mas (calon ikan semang) yang mampu menerima sel donor dengan baik dan memiliki keberhasilan kolonisasi yang tinggi. Testis ikan neon tetra didisosiasi menggunakan larutan tripsin 0,5%. Sel testikular diwarnai dengan PKH-26, kemudian ditransplantasikan ke rongga peritoneal benih ikan mas umur tujuh, sepuluh, dan 14 hari setelah menetas. Hasil penelitian menunjukkan bahwa tingkat kelangsungan hidup ikan mas perlakuan transplantasi umur tujuh hari (31,25%) lebih rendah dibandingkan dengan perlakuan transplantasi umur sepuluh hari (37,50%) dan 14 hari (56,25%). Persentase ikan terkolonisasi sel donor pada hari ke-21 pascatransplantasi pada benih umur tujuh dan sepuluh hari adalah sama (80%), sedangkan transplantasi benih umur 14 hari sebesar 60%. Berdasarkan keberhasilan transplantasi secara kumulatif (tingkat kelangsungan hidup dan kolonisasi), transplantasi pada benih umur 14 hari (33,75%) menunjukkan hasil lebih tinggi dibandingkan dengan perlakuan transplantasi pada benih umur tujuh hari (25,00%) dan benih umur sepuluh hari (30,00%). Transplantasi sel testikular ikan neon tetra pada benih ikan mas telah berhasil dilakukan, dan umur optimum benih ikan mas adalah 14 hari setelah menetas. Kata kunci: transplantasi, kolonisasi, sel testikular, ikan mas, ikan neon tetra 

Frequency and persistency of DNA vaccine encoding GP25 by oral on common carp

Jurnal Akuakultur Indonesia Vol 12, No 2 (2013): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT Koi herpesvirus (KHV) is a major viral pathogen that infects common carp and koi. KHV disease outbreak is happened in almost all centre of common carp culture in Indonesia and caused mass mortality. Deoxyribonucleic acid (DNA) vaccination method is one of ways to cope with KHV infection. Vaccines were commonly given by injection. The aim of this research was to get frequency and persistency of DNA vaccine encoding GP25 given by oral delivery method in common carp. This research would like to determine dose, frequency of vaccination, persistency of DNA vaccine and culture medium for the bacterial host. DNA vaccine persistency test was done by using polymerase chain reaction (PCR) method with the specific primer for GP25 gene. The results showed that level of DNA vaccine that could be detected in feed was 7.56 ng (equal to 1.598×1010 copies). Efficient culture medium for Escherichia coli DH5α carrying DNA vaccine was LB triptone. Feeding fish with diet supplemented with 1 mL E. coli DH5α containing DNA vaccine for each fish and two times a week allowed persistence of DNA vaccine in kindney and spleen. Keywords: common carp, KHV, DNA vaccine, GP25, persistance  ABSTRAK Koi herpesvirus (KHV) adalah virus patogen utama yang menginfeksi ikan mas dan ikan koi. Wabah penyakit KHV terjadi di hampir semua sentra budidaya ikan mas di Indonesia dan menyebabkan kematian massal ikan. Metode vaksinasi DNA merupakan salah satu cara yang dapat dilakukan untuk menanggulangi serangan KHV. Pemberian vaksin umumnya dilakukan dengan cara injeksi. Tujuan penelitian ini adalah untuk menguji frekuensi dan persistensi vaksin DNA GP25 antivirus KHV yang diberikan melalui oral pada ikan mas. Pada penelitian ini dilakukan uji dosis, frekuensi pemberian vaksin, persistensi vaksin DNA, dan media kultur bakteri inang. Persistensi vaksin DNA dianalisis menggunakan metode PCR dengan primer spesifik gen GP25. Hasil penelitian menunjukkan bahwa dosis vaksin DNA yang dapat terdeteksi dalam pakan adalah 7,56 ng (setara dengan 1,598×1010 copy). Media kultur yang efisien bagi bakteri Escherichia coli DH5α pembawa vaksin DNA adalah LB tripton. Pemberian pakan bervaksin berupa bakteri konsentrasi 1 mL/ekor ikan dengan frekuensi dua kali seminggu menghasilkan persistensi DNA GP25 di ginjal dan limpa. Kata kunci: ikan mas, KHV, vaksin DNA, GP25, persistensi 

Electroporation and GFP-labelled transplantation of testicular cells in Nile tilapia

Jurnal Akuakultur Indonesia Vol 12, No 2 (2013): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT Transplantation technology can be applied to generate fish surrogate broodstock. A donor germinal cells that was used in transplantation are labeled to distinguish it with endogenous cells. Donor cells are generally derived from transgenic fish carrying a marker or cells labeled by PKH-26. This study was performed to obtain an alternative method of cell labelling using electroporation. Testicular cells were taken from 4-months old Nile tilapia as a model. Electroporation was performed with testicular cell density of 104 cells/µL, pJfKer-GFP concentration of 50 ng/µL, and a pulse length of 20 ms at 0, 100, 200, and 300 volts. At amount of 5x103 cells/0.5µL electroporated testicular cells were then injected into the intraperitoneal cavity of 3-day-old Nile tilapia larva. The results showed that survival of the electroporated cells of 100 and 200 volt-treatments was similar (P>0.05), and higher than 300 volt (P<0.05). Number of fluorescent cells was not significantly different among treatments. The highest cell colonization in transplanted fish was obtained in 200-volt treatment (66.67%). As conclusion, 200-volt electroporation with was a suitable tool to label testicular cells for transplantation. Keyword: electroporation, GFP, label, Nile tilapia, transplantation  ABSTRAK Teknologi transplantasi merupakan suatu teknologi yang dapat menghasilkan induk pengganti. Sel donor berupa sel germinal yang akan digunakan dalam transplantasi diberi label agar dapat dibedakan dengan sel resipien. Umumnya sel donor diperoleh dari ikan transgenik yang membawa marka atau diwarnai dengan PKH-26. Penelitian ini bertujuan untuk mencari metode alternatif dalam pemberian label pada sel dengan elektroporasi. Sel testikular diperoleh dari ikan nila berumur empat bulan sebagai model. Elektroporasi dilakukan dengan kepadatan sel 104 sel/µL, konsentrasi pJfKer-GFP 50 ng/µL, dan panjang kejut 20 ms pada 0, 100, 200, dan 300 volt. Sebanyak 5x103 sel dalam 0,5 µL larutan hasil elektroporasi disuntikkan ke dalam rongga intraperitoneal larva berumur tiga hari setelah menetas. Hasil penelitian menunjukkan bahwa kelangsungan hidup sel pada kejut 100 dan 200 volt tidak signifikan (P<0,05), akan tetapi lebih tinggi bila dibandingkan dengan perlakuan 300 volt (P<0,05). Jumlah sel yang berpendar tidak berbeda antara perlakuan. Persentase kolonisasi sel pada ikan hasil transplan tertinggi pada perlakuan 200 volt (66,67%). Kesimpulan yang diperoleh dari hasil penelitian adalah perlakuan elektroporasi dengan kejut listrik 200 volt dapat digunakan untuk memberi label pada sel testikular yang akan ditransplantasikan. Kata kunci : elektroporasi, GFP, label, ikan nila, transplantasi 

Potential transmission test of GP25 vaccine in normal flora bacteria of common carp culture media

Jurnal Akuakultur Indonesia Vol 14, No 1 (2015): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT Koi herpesvirus (KHV) is a virus that infects common carp and koi Cyprinus carpio. KHV attacks most stadia of common carp and koi and causes mortality up to 80–95% of the population. One way to prevent the spread of KHV is  by applying DNA vaccine. This research was conducted to test potential transmission of DNA vaccine encoding glycoprotein 25 (vaccine GP25) toward normal flora bacteria of media cultivation of common carp. Bacteria was isolated from pond water of common carp and tested for their sensitivity to ampicillin. Research was performed by adding vaccine GP25 to ampicillin-sensitive bacteria at a dose of 12.5 µg/100 µL and incubated at 28 °C for 30, 60, 180, and 300 minutes then plated on media containing ampicillin. The grown bacteria cells were tested for the existence of plasmid bearing gen GP25 through bacteria colony cracking. The results of this research showed that there was no bacteria contained plasmid bearing gen GP25. Keywords: DNA vaccines, common carp, vaccine safety  ABSTRAK Koi herpesvirus (KHV) merupakan virus yang menginfeksi ikan mas dan koi Cyprinus carpio. KHV menyerang hampir semua stadia ikan mas dan koi dan menyebabkan kematian hingga 80–95% dari populasi. Salah satu cara penanggulangan penyebaran KHV adalah dengan penggunaan vaksin DNA. Penelitian ini dilakukan untuk menguji potensi transmisi vaksin DNA glikoprotein 25 (vaksin GP25) pada bakteri flora normal pada media budidaya ikan mas. Bakteri diisolasi dari kolam budidaya ikan mas dan diuji sensitivitasnya terhadap antibiotik ampisilin. Penelitian dilakukan dengan menambahkan vaksin GP25 ke larutan bakteri sensitif ampisilin dengan dosis 12,5 µg/100 µL dan diinkubasi pada suhu 28 °C selama 30, 60, 180, dan 300 menit kemudian disebar pada media mengandung antibiotik ampisilin. Sel bakteri yang tumbuh diuji untuk mendeteksi keberadaan plasmid pembawa gen GP25 dengan menggunakan metode seleksi koloni bakteri (colony cracking). Hasil penelitian ini menunjukkan bahwa tidak ada bakteri yang mengandung plasmid pembawa gen GP25. Kata kunci: keamanan vaksin, ikan mas, vaksin DNA

Performance of broodstock and hybrid juvenile of Egyptian and sangkuriang Clarias gariepinus strains

Jurnal Akuakultur Indonesia Vol 14, No 2 (2015): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT This study was conducted to evaluate reproduction of broodstock and intraspecific hybrid juvenile performance of Egyptian (M) and sangkuriang (S) strain African catfish Clarias gariepinus at nursery phase. Intraspecific hybridization of African catfish was carried out reciprocally (SM and MS) and purebreed (SS and MM), each was with three replicates. Fish spawning was conducted by artificial fertilization, and larvae were reared at 1st, 2nd and 3rd nursery phases, subsequently. The results showed that broodstock performance (male and female) of both strains were not significantly different (P>0.05) on all reproduction traits, except female’s gonadosomatic index. Fertilization and hatching rates of all hybrids were not significantly different (P>0.05). MM juvenile had higher growth performances than other juvenile hybrids. Heterosis of total length, standard length, and body weight were varied, whereas the survival showed positive heterosis. SM juvenile showed positive growth heterosis in 3rd nursery phase (total length, standard length, and body weight were 2.61%; 2.16%, and 4.79% respectively). Survival heterosis of MS juvenile (24.20% for total length; 103.13% for standard length and 11.62% body weight) was higher than SM juvenile (6.86%; 48.57%, and 3.09%) on all nursery phases Keywords: African catfish, intraspecific hybridization, growth, survival, heterosis  ABSTRAK Penelitian ini dilakukan untuk menguji performa reproduksi induk dan benih hasil hibridisasi intraspesifik ikan lele Afrika Clarias gariepinus strain Sangkuriang (S) dan Mesir (M) pada fase pendederan. Hibridisasi intraspesifik ikan lele Afrika dilakukan secara resiprokal (SM dan MS) dan galur murni (SS dan MM) masing-masing dengan tiga ulangan. Pemijahan dilakukan secara buatan dan larva yang dihasilkan dipelihara pada fase pendederan pertama, kedua, dan ketiga. Hasil penelitian menunjukkan bahwa performa induk ikan lele jantan dan betina pada kedua strain tidak berbeda nyata (P>0,05) pada seluruh parameter reproduksi, kecuali indeks gonadosomatik (GSI) pada induk betina. Derajat pembuahan dan penetasan telur pada seluruh persilangan tidak berbeda nyata (P>0,05). Pertumbuhan benih persilangan MM lebih tinggi, dibandingkan benih hasil persilangan lain. Nilai heterosis panjang total, panjang standar, dan bobot tubuh yang dihasilkan pada setiap stadia pendederan bervariasi, sedangkan nilai heterosis kelangsungan hidup menunjukkan hasil seluruhnya positif dibandingkan rataan galur murni. Heterosis pertumbuhan benih persilangan SM pada pendederan ketiga menunjukkan hasil yang positif (2,61% untuk panjang total; 2,16% untuk panjang standar dan 4,79% untuk bobot tubuh). Nilai heterosis kelangsungan hidup benih persilangan MS (24,20% untuk panjang total; 103,13% untuk panjang standar dan 11,62% untuk bobot tubuh) lebih tinggi dibandingkan benih persilangan SM (6,86%, 48,57% dan 3,09%) pada setiap stadia pendederan. Kata kunci: ikan lele Afrika, hibridisasi intraspesifik, pertumbuhan, kelangsungan hidup, heterosis 

The sex ratio and testosterone levels in tilapia immersed in different doses of 17α-methyltestosterone

Jurnal Akuakultur Indonesia Vol 16, No 1 (2017): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

  Tilapia fish farming using monosex male population has been reported to have 10% higher productivity compared to mix-sex culture. This study aimed to determine immersion dose of 17α-metiltestosterone (MT) that allowed higher male percentage, growth, survival, and measure testosterone level in fish body. The experiment was designed as three immersion doses, namely: 0; 1.8; and 5.4 mg/L MT, each with three replications. Immersion was conducted to 14-day-old larvae for four hours, with density of 100 fish/L of water. Testosterone levels in fish was measured using ELISA method, and sex identification was performed by histology method. The result showed that percentage of male fish was the same in the treatment of 1.8 mg/L and 5.4 mg/L, and 53–65% higher than the control without MT treatment (54% male). Growth and survival of fish until day 56 post immersion were the same. By ELISA analysis, the levels of testosterone in larvae just after immersion was similar in 1.8 mg/L and 5.4 mg/L treatments, decreased drastically on day 14 after immersion, and then the levels of testosterone to be similar with the control on day 28, i.e. about 2 ng/g. By PCR method with specific primer, sex reversed and normal males could be distinguished, and on day 71 the testosterone levels in sex reversed and normal males fish was also the same, 0.3 ng/g (P>0.05). Thus, sex reversal by immersion at a dose of 1.8 mg/L can be consider as a standard protocol for monosex tilapia production. Testosterone level in the body of MT-treated fish becomes the same to the control of less than one month post immersion

Aromatase gene expression and masculinization of Nile tilapia immersed in water 36 °C containing 17α-methyltestosterone

Jurnal Akuakultur Indonesia Vol 16, No 1 (2017): Jurnal Akuakultur Indonesia
Publisher : ISSA

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Abstract

ABSTRACT  Immersion of undifferentiated larval tilapia in high temperature and 17α-methyltestosterone (MT) cab increase the male ratio. However, the effectiveness of immersion in high temperature of water containing MT remains to be evaluated. The purposes of this study were: 1) evaluate the male ratio, growth, and survival of tilapia, and 2) analyze the aromatase brain-type gene expression level in tilapia after immersing in high temperature (36 °C) containing MT at 2 mg/L for four hour with single and double immersion. Aromatase gene expression was analyzed by semi-quantitative RT-PCR (sqRT-PCR) method. The result showed that higher monosex male ratio was obtained by single immersion of MT at 36 °C at room temperature. Gene expression level of aromatase brain-type was lower on single immersion and increased significantly at second immersion compared to control (immersion at room temperature without MT). Immersion using MT and high temperature had no significant effect on fish survival. However the specific growth rate and fish biomass were higher than control. Thus, monosex male tilapia can be produced by single immersion of undifferentiated larvae at 36 °C temperature containing MT. Keywords: male ratio, aromatase, Oreochromis niloticus, temperature, 17α-methyltestosterone  ABSTRAK  Perendaman larva ikan nila yang belum terdeferensiasi kelaminnya dengan suhu tinggi dan hormon 17α-metiltestosteron (MT) dapat meningkatkan nisbah kelamin jantan. Tetapi, efektivitas perendaman menggunakan MT pada suhu tinggi belum diteliti. Tujuan dari penelitian ini adalah 1) mengevaluasi nisbah kelamin jantan, pertumbuhan, dan kelangsungan hidup ikan nila, dan 2) menganalisis ekspresi gen aromatase tipe-otak pada ikan direndam menggunakan MT dengan dosis 2 mg/L selama empat jam sebanyak satu dan dua kali perendaman pada suhu 36 °C. Ekspresi gen aromatase dianalisis menggunakan metode RT-PCR semi-kuantitatif (sqRT-PCR). Hasil penelitian menunjukkan bahwa kombinasi perendaman MT satu kali pada suhu 36 °C lebih tinggi menghasilkan ikan nila jantan monoseks dibandingkan perendaman MT satu kali pada suhu ruang. Tingkat ekspresi gen aromatase tipe otak pada perendaman satu kali lebih rendah, dan meningkat secara signifikan pada perendaman kedua dibandingkan dengan kontrol (perendaman pada suhu ruang tanpa MT). Perendaman larva menggunakan MT dan suhu 36 °C tidak berpengaruh nyata terhadap kelangsungan hidup, tetapi laju pertumbuhan spesifik dan biomassa ikan perlakuan tersebut lebih tinggi daripada kontrol. Dengan demikian, ikan nila jantan monoseks dapat diproduksi dengan perendaman satu kali pada larva yang belum terdeferensiasi jenis kelaminnya menggunakan MT pada suhu 36 °C. Kata kunci: rasio jantan, aromatase, Oreochromis niloticus, suhu, 17α-metiltestosteron