Andi Tenriulo
Balai Penelitian dan Pengembangan Budidaya Air Payau, Maros, Sulawesi Selatan

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PENGARUH APLIKASI DSRNA VP-15 IN VITRO DAN IN VIVO TERHADAP SINTASAN DAN RESPONS IMUN UDANG WINDU PENAEUS MONODON Parenrengi, Andi; Tenriulo, Andi; Mulyaningrum, Sri Redjeki Hesti; Lante, Samuel; Nawang, Agus
Jurnal Riset Akuakultur Vol 14, No 4 (2019): (Desember, 2019)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jra.14.4.2019.213-223

Abstract

Teknologi RNA interference (RNAi) merupakan salah satu pendekatan yang digunakan untuk meningkatkan resistensi udang windu terhadap infeksi patogen termasuk WSSV. Pengembangan teknologi RNAi melalui aplikasi untai ganda RNA (dsRNA) yang berasal dari gen pengkode viral protein (VP) dari WSSV telah mulai dikembangkan pada udang. Penelitian ini bertujuan untuk mengkaji sintasan dan respons imun udang windu yang diberi VP-15 pasca uji tantang dengan WSSV. Udang windu (panjang 15,21 ± 1,19 cm dan bobot 32,5 ± 1,83 g) diinjeksi dengan 0,2 µg/ekor dsRNA in vitro (A), dsRNA in vivo (B), dan larutan garam/kontrol (C). Setelah tiga hari vaksinasi, udang windu ditantang dengan WSSV dengan dosis 50 µL/ekor. Pengamatan sintasan dilakukan setiap hari, sedangkan respons imun (THC dan aktivitas proPO) dilakukan pada awal dan hari ke-1, ke-3, dan ke-5 pasca uji tantang, serta analisis ekspresi gen antivirus dan histopatologi hepatopankreas dilakukan pada akhir penelitian. Hasil penelitian menunjukkan bahwa aplikasi dsRNA berpengaruh nyata (P<0,05) terhadap sintasan, THC, dan proPO. Sintasan udang windu yang diberi dsRNA VP-15 in vitro dan in vivo memberikan sintasan yang lebih tinggi 75% dibandingkan dengan kontrol. Nilai proPO tertinggi didapatkan pada dsRNA in vivo (0,138); kemudian dsRNA in vitro (0,093); dan terendah kontrol (0,061); sedangkan THC tertinggi (5.704 x 104 sel/mL) pada dsRNA in vivo, kemudian dsRNA in vitro (3.516 x 104 sel/mL) dan terendah pada perlakuan kontrol (3.322 x 104 sel/mL). Ekspresi gen antivirus semakin meningkat dengan semakin lamanya udang windu terpapar dengan WSSV. Jaringan hepatopankreas udang windu pada perlakuan kontrol (tanpa dsRNA) menunjukkan adanya kerusakan sel akibat infeksi virus.RNA interference (RNAi) technology is one of the approaches used to improve tiger shrimp Penaeus monodon resistance against WSSV infection. The development of RNAi technology through double-stranded RNA (dsRNA) isolated from gene encoding viral protein (VP) of WSSV has been applied to shrimp. This study was aimed to assess the survival rate and immune response of injected-VP-15 WSSV tiger shrimp after a challenge with WSSV. The tiger shrimp (15.21 ± 1.19 cm in length and 32.5 ± 1.83 g in weight) were injected with 0.02 µg/shrimp of in vitro dsRNA (A), in vivo dsRNA (b) and saline solution (C). After three days of vaccination, the tiger shrimp were challenged with WSSV using a dosage of 50 µL/shrimp. The survival rate was observed daily. Analyses of immune responses (hemocyte total and PO activity) were performed in several stages: before the challenge test and day-1, day-3, and day-5 post-challenge test. The expression of the antivirus gene and hepatopancreas histophatology were was observed at the end of the experiment. The results showed that the application of dsRNA significantly influenced the shrimp survival rate, THC, and proPO. Tiger shrimp injected with dsRNA VP-15 of in vitro and in vivo exhibited a higher 75% survival rate than the control (P<0.05). The highest proPO activity (0.138) was obtained at dsRNA in vivo, followed by dsRNA in vitro (0.093) and the lowest (0.061) in the control. The highest THC (5,704 x 104 cell/mL) was in vivo dsRNA, then in vitro dsRNA (3,516 x 104 cell/mL), and the lowest in the control (3,322 x 104 cell/mL). The longer the exposure with WSSV, the higher the antivirus gene expression. Histopathology analysis showed some damages to the hepatopancreas cells in the control shrimp (without dsRNA) caused by the virus infection.
CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798) Parenrengi, Andi; Alimuddin, Alimuddin; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 12, No 2 (2017): (December, 2017)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (157.395 KB) | DOI: 10.15578/iaj.12.2.2017.67-75

Abstract

White spot syndrome virus (WSSV) has caused mass mortality on tiger shrimp (Penaeus monodon) culture and adversely affects prawn industry worldwide including Indonesia. It is well known that the protein structure of WSSV plays an important role in the virus infection and morphogenesis process. A viral protein structure called VP-15 is located in the nucleocapsid of virion virus. The protein structure involves in the life cycle of WSSV in host cells. A gene encoding VP-15 could be involved in constructing the RNA interference (RNAi), so it is needed to isolate and characterize for RNAi technology purpose. The study was aimed to isolate and characterize the VP-15 from the infected WSSV tiger shrimp. The characterization of VP-15 was undertaken through assessment of nucleotide sequence, amino acid deduction, alignment nucleotide/protein searches using Genetyx and BLAST program, and dendrogram construction analysis. The results showed that VP-15 was successfully isolated in form of ORFDNA with a fragment size of 243 bp. The phylogenetic tree analysis revealed three clusters corresponding to the time (year) of isolates collection. The VP-15 consisted of 80 amino acids, two start codons (ATG), one stop codon (TAA), and one Kozak context (AAAATGG). Hydrophilic amino acid was the highest composition (44.2%), followed by neutral (31.2%) and hydrophobic (24.6%) amino acid groups. The VP-15 was rich in amino acid of lysine (21.3%), arginine (22.9%) and serine (24.6%). The successful isolation of VP-15 is a very important step in providing a basic yet suitable material in constructing the dsRNA vaccine to control shrimp diseases in aquaculture.
EXPRESSION OF ANTIVIRAL GENE ON TIGER SHRIMP Penaeus monodon AT DIFFERENT TISSUE AND BODY SIZE Parenrengi, Andi; Tenriulo, Andi; Lante, Samuel
Indonesian Aquaculture Journal Vol 7, No 2 (2012): (December 2012)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (484.161 KB) | DOI: 10.15578/iaj.7.2.2012.95-104

Abstract

The role of tiger shrimp defense against invading pathogen on molecular level such antiviral gene expression is limited to be reported. Gene expression is a process which codes information of genes that is converted to the protein as a phenotype. Distribution of PmAV antivirus gene, that has been reported as an important gene on non-specific response immune, is needed to be observed to several organs/tissues and size of tiger shrimp. The aim of this study is to determine the distribution of gene antiviral expression at several organ/tissue and size of shrimp. The organs/tissues observed in this study were: gill, hepatopancres, muscle tissue, eyes, heart, stomach, gonad, and intestine. While the size of shrimp consisted of three groups, those are: (A) 10-20 g/ind., (B) 30-40 g/ind., and (C) 60-70 g/ind. Analysis of antiviral gene expression was performed by RNA extraction, followed by the cDNA syntesis, and amplification of gene expression by semi-quantitative PCR. The result of PCR optimation showed the optimal concentration of cDNA and primer was 1 μL and 50 mol, respectively for PCR final volume of 25 μL. Antiviral gene was expressed on the hepatopancreas and stomach in percentage of 50.0% and 16.7%, respectively. While the highest percentage of individual expressing the antiviral gene was observed in the shrimp size of C (66.7%), followed by B (50.0%) and A (16.7%). The result of study implied that the hepatopancreas has importantly involed in tiger shrimp defense mechanism on viral infection.
ANALYSIS OF IMMUNE RESPONSES ON TRANSGENIC TIGER SHRIMP (Penaeus monodon) AGAINST PATHOGENIC BACTERIUM Vibrio harveyi Parenrengi, Andi; Tampangallo, Bunga Rante; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 9, No 1 (2014): (June 2014)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (406.087 KB) | DOI: 10.15578/iaj.9.1.2014.23-32

Abstract

Vibriosis is one of main diseases of the black tiger shrimp Penaeus monodon infected by pathogenic bioluminous bacterium Vibrio harveyi that can cause mass mortalities in shrimp culture. The bacteria can also trigger the disease white spot syndrome virus (WSSV). An effort to produce shrimp disease-resistant strains has been done through transgenesis technology with antiviral gene transfection. By this technology, it is expected an increase in the immune response of shrimp in a variety of diseasecausing pathogens. This study aimed to determine the immune responses (total haemocytes, haemocyte differentiation, and phenoloxydase activity) of transgenic tiger shrimp against pathogenic bacterium V. harveyi. Research using completely randomized design, which consists of two treatments and three replications. Test animals being used were transgenic and non-transgenic shrimp with size, weight 3.93±1.25 g and a total length of 7.59±0.87 cm. Treatments being tested were the injection of bacterium V. harveyi (density of 5x106 cfu/mL) of 0.1 mL/individual on transgenic (A) and non-transgenic shrimp (B). Immune response parameters such as total haemocytes, haemocyte differentiation, and phenoloxydase activity were observed on day 1, 3, and 6 days after challenging. Data were analyzed using t-test by SPSS software. The results showed that the total haemocyte of transgenic shrimp was not significantly different (P>0.05) from non-transgenic shrimp, but haemocyte differentiation and phenoloxydase activity were significantly different (P<0.05) especially on sixth days after being exposed to the bioluminescent bacteria. The study results implied that transgenic shrimp has a better immune response compared than non-transgenic shrimp.
IN VITRO GROWTH RATE OF Kappaphycus alvarezii MICROPROPAGULE AND EMBRYO BY ENRICHMENT MEDIUM WITH SEAWEED EXTRACT Suryati, Emma; Rosmiati, Rosmiati; Parenrengi, Andi; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 10, No 1 (2015): (June 2015)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (416.68 KB) | DOI: 10.15578/iaj.10.1.2015.13-17

Abstract

The development of micropropagule and embryo of seaweed depend on nutrient and fertilizer used. Seaweed has been reported contain hormone regulators such as auxine, cytokinine, gibbereline, and various minerals applied in stimulating the growth ocra plant and wheat culture. The objectives of this study were to determine the potential of Kappaphycus alvarezii extract and its optimal concentration in accelerating of Kappaphycus alvarezii micropropagule and embryo growth. Micropropagule and embryo produced through callus induction were planted into PES 1/20 liquid medium supplemented with seaweed extract at the concentrations of 0 (control), 25, 50, 75, and 100 μL in 10 mL of medium. The results showed that medium enrichment with 50 μL of seaweed extract had the highest survival rate and growth of thallus. In addition, this concentration was also resulted in a good performance of K. alvarezii thallus with the lighter color. The advantage of this study for seaweed cultivation in Indonesia, among others, seaweed can be used as fertilizer, especially in the maintenance of seaweed seed, so that cultivation can be better develop.
PENGARUH PEMBERIAN EKSTRAK PAKIS SEBAGAI MOULTING STIMULAN PADA INDUK UDANG WINDU (Penaeus monodon. Fab) DI HATCHERY Suryati, Emma; Tenriulo, Andi; Tonnek, Syarifuddin
Jurnal Riset Akuakultur Vol 8, No 2 (2013): (Agustus 2013)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (305.561 KB) | DOI: 10.15578/jra.8.2.2013.221-228

Abstract

Tumbuhan pakis (Pteridophyta) merupakan salah satu tumbuhan yang memilikikandungan senyawa steroid yaitu fitoekdisteroid dalam bentuk 20-Hydroxyecdyson atau Ecdysteron berfungsi sebagai moulting stimulan pada krustase. Pada umumnya ecdysteron ditemukan pada krustase baik yang ada di darat maupun yang berada di dalam air seperti kepiting, udang, dan krustase lainnya yang ditemukan secara alami dan berfungsi sebagai pengatur proses penggantian kulit dan mengontrol pembentukan exoskeleton baru untuk menggantikan exoskeleton yang lama. Selain ablasi proses moulting pada udang dapat diinduksi melalui penambahan 20-hydroksi ecdysteron (20 E) pada hemolim sehingga fase premoulting dapat diatur sesuai dengan kebutuhan. 20-hydroksiecdysteron dapat diperoleh dari ekstrak tumbuhan di antaranya bayam, asparagus, pakis, dan lain-lain melalui pemisahan dengan ekstraksi, fraksinasi, dan pemurnian dengan HPLC dilanjutkan dengan elusidasi struktur. Pemberian ekstrak pakis pada induk udang windu untuk memacu terjadinya pergantian kulit dilakukan melalui dengan beberapa konsentrasi menggunakan desain Rancangan Acak Lengkap (RAL). Hasil analisis memperlihatkan bahwa kandungan phytoecdysteron pada pakis perkisar 230-730 mg/L dari larutan ekstrak yang setara dengan 20 g bahan segar. Konsentrasi ECD 25 mg/L, memperlihatkan respon yang paling baik sebagai moulting stimulan.
CLONING OF ProAV PROMOTER ISOLATED FROM TIGER PRAWN Penaeus monodon Parenrengi, Andi; Alimuddin, Alimuddin; Sukenda, Sukenda; Sumantadinata, Komar; Yamin, Muhammad; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 4, No 1 (2009): (June 2009)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (143.086 KB) | DOI: 10.15578/iaj.4.1.2009.1-7

Abstract

Promoter is a specific DNA sequence involved in the transcription of a particular gene. It is usually located in the upstream of the gene they regulate. Isolation and characterization of promoter is essentially needed in order to establish the sequence analysis and transcription factor that are used in the regulation of gene expression. The research was conducted to analyze the characteristics of Penaeus monodon anti viral gene promoter (ProAV) towards generation of auto-transgenic tiger prawn, P. monodon. ProAV promoter was isolated by PCR (Polymerase Chain Reaction) method and the purified DNA fragment was cloned into pGEM-T Easy cloning vector. The promoter sequence was characterized by using BLAST-N and Genetyx version 7 softwares. The results showed the success in isolating a promoter from tiger prawn of 368 bp in length. BLAST-N analysis showed that the sequence of isolated promoter has high similarity (95%-98%) compared to the other promoters in the GeneBank. The study revealed the existence of important transcription factors (TATA box, MRE, TCF-1, and other potential regulatory elements) are identified in the promoter sequence.
KERAGAMAN MORFOLOGI UDANG PAMA ( Penaeus semisulcatus ) DARI PERAIRAN SULAWESI SELATAN DAN SULAWESI TENGGARA Parenrengi, Andi; Sulaeman, Sulaeman; Hadie, Wartono; Tenriulo, Andi
Jurnal Riset Akuakultur Vol 2, No 1 (2007): (April 2007)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (174.118 KB) | DOI: 10.15578/jra.2.1.2007.27-32

Abstract

Udang pama, Penaeus semisulcatus merupakan salah satu jenis krustase lokal yang memiliki prospek untuk dikembangkan sebagai kandidat spesies budi daya tambak. Penelitian ini bertujuan untuk mengetahui keragaman morfologi dan jarak genetik udang pama yang berasal dari Sulawesi Selatan dan Sulawesi Tenggara. Principle component analysis (PCA) dan discriminant analysis digunakan untuk mengetahui keragaman morfologi antar ketiga populasi alami udang pama. Hasil penelitian menunjukkan bahwa morfologi udang pama dari Munte dan Lampia (Sulawesi Selatan) berbeda dengan udang pama yang berasal dari Kassipute (Sulawesi Tenggara). Analisis kluster juga mengindikasikan adanya dua kluster utama, di mana kluster pertama merupakan gabungan antara udang pama dari Munte dan Lampia, sedangkan kluster lainnya adalah udang pama yang berasal dari Kassipute. Jarak genetik yang didapatkan memperlihatkan kekerabatan terdekat adalah udang pama yang berasal dari MunteLampia (5,424) dan terjauh pada udang pama yang berasal dari Lampia-Kassipute (48,350).Green tiger prawn, Penaeus semisulcatus is one of the prospective local crustaceans as a candidate species of shrimp pond culture. The objective of this study is to reveal the morphology diversity and genetic distance of green tiger prawn from South Sulawesi and Southeast Sulawesi. Principle component analysis (PCA) and discriminant analysis were used to analyze morphometric variations among the three natural populations. Result showed that the morphology of green tiger prawn from Munte dan Lampia (South Sulawesi) was relatively different with prawn collected from Kassipute (Southeast Sulawesi). Cluster analysis also indicated the existing of two main clusters i.e. green tiger prawn from Munte and Lampia as the first cluster and Kassipute as the second cluster. The lowest value of genetic distance was obtained from Munte-Lampia (5.424) and the highest genetic distance was obtained from Lampia-Kassipute (48.350).
GENETIC VARIABILITY AND POPULATION STRUCTURE OF GROUPER (Epinephelus suillus) FROM MAKASSAR STRAIT AND BONE BAY, SOUTH SULAWESI, INDONESIA Parenrengi, Andi; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 3, No 2 (2008): (December 2008)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (455.808 KB) | DOI: 10.15578/iaj.3.2.2008.77-87

Abstract

Random Amplified Polymorphic DNA (RAPD) was employed to determine the genetic variability and population structure of grouper (Epinephelus suillus) from Makassar Strait and Bone Bay, South Sulawesi, Indonesia. Genomic DNA was isolated from preserved muscle tissue using Phenol-Chloroform technique. Among 24-screened arbitrary primers, ten primers (OPA-02, OPA-06, OPA-08, OPA-10, OPA-15, OPA-16, OPA-17, OPA-18, OPA-19 and CA-05) were selected to generate RAPD fingerprinting of grouper populations. The ten primers generated a total of 212 fragments (loci) and 120 polymorphic fragments in their size ranging from 250 to 2,500bp. The high polymorphism (60%) was obtained from Makassar population followed by Bone (59%) and Pare-Pare populations (50%). Similarity index of individuals was 0.86±0.07 for Pare-Pare, 0.80±0.11 for Makassar and 0.82±0.07 for Bone population. Fifteen fragments from ten primes were identified as species-specific markers of E. suillus. The UPGMA cluster analysis showed that the dendrogram seemed to be clustered according to their geographical location, where Pare-Pare population was genetically closer to Makassar population (D=0.20) than to Bone population (D=0.24).
GENETIC VARIABILITY OF THREE POPULATIONS OF FLYING FISH, Hirundichthy oxycephalus FROM MAKASSAR STRAIT Parenrengi, Andi; Tenriulo, Andi; Ali, Syamsul Alam
Indonesian Aquaculture Journal Vol 7, No 1 (2012): (June 2012)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (470.605 KB) | DOI: 10.15578/iaj.7.1.2012.1-10

Abstract

Flying fish, Hirundichthy oxycephalus is one of economically important marine species to Indonesia, particularly in Makassar Strait and Flores Sea. However, there is a limited published data on genetic variation in molecular marker level of this species. Random Amplified Polymorphic DNA (RAPD) was employed in this study to determine the genetic variability of three populations of flying fish collected from Takalar, Pare-Pare, and Majene in Makassar Strait. Genomic DNA was isolated from preserved muscle tissue using phenol-chloroform technique. Two selected arbitrary primers (CA-01 and P-40) were performed to generate RAPD finger printing of flying fish populations. The two primers generated a total of 81 fragments (loci) and 50 polymorphic fragments with size ranging from 125 to 1,250 bp. There were no significant differences in number of fragment and number of polymorphic fragment among populations. The high polymorphism (63.5±7.4%) was obtained from Takalar population followed by Pare-Pare (58.3±19.6%) and Majene population (57.7±0.8%). Similarity index of individuals was 0.60±0.17 for Takalar, 0.63±0.17 for Majene and 0.75±0.21 for Pare-Pare population. Seven fragments were identified as species-specific markers of H. oxycephalus. The UPGMA cluster analysis showed that the Takalar population was genetically closer to Pare-Pare population (D= 0.0812) than to Majene population (D= 0.1873).