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Pengaruh Bioenkapsulasi Artemia salina Terhadap Tingkat Kelangsungan Hidup Benih Ikan Nila (Oreochromis niloticus) Eko Suyanto; Yasir Saifur Rahman; Murwantoko Murwantoko
Biotropika: Journal of Tropical Biology Vol 7, No 2 (2019)
Publisher : University of Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.biotropika.2019.007.02.5

Abstract

Permintaan ikan nila (Oreochromis niloticus) semakin meningkat dari tahun ke tahun namun terkendala dalam ketersediaan benih karena tingginya resiko serangan penyakit streptococcosis yang menurunkan kelangsungan hidup (sintasan) benih. Tujuan penelitian ini adalah adalah untuk mengetahui pengaruh pemberian pakan bioenkapsulasi Artemia salina dengan Spirulina platensis terhadap sintasan benih ikan nila yang telah diinjeksi bakteri Streptococcus sp. dalam upaya mencegah penyakit streptococcosis. Bioenkapsulasi dilakukan dengan cara kultur A. salina diberi pakan tepung S. platensis ukuran 37 mikron selama 5 jam lalu dipanen dan disimpan pada suhu 4oC. Bioenkapsulan dianalisis proksimat meliputi kadar air, kadar protein total, kadar lemak, kadar abu, kadar serat kasar dan kadar karbohidrat sedangkan analisis kadar asam lemak dianalisis menggunakan Gas chromatography. Perlakuan pakan benih ikan nila yaitu bioenkapsulan dan pellet ikan dengan perbandingan variasi 25%:75% (BASP1), 50%:50% (BASP2), 75%:25% (BASP3), 100%:0% (BASP4) dan 0%:100% (K) lalu diuji tantang dengan bakteri Streptococcus sp konsentrasi 106 CFU/mL secara rendaman. Pakan bioenkapsulasi A. salina dengan S. platensis memberikan pengaruh terhadap tingkat kelangsungan hidup benih ikan nila. Peningkatan kelangsungan hidup terbaik diperoleh pada hari ke-7 sebesar 26,7%. Pemberian pakan bioenkapsulan dengan konsentrasi 50-75% cukup efektif diberikan hingga hari ke-14 untuk meningkatkan sintasan benih ikan nila dalam usaha pencegahan penyakit streptococcosis. Semakin tinggi konsentrasi pakan bioenkapsulan akan meningkatkan resiko kematian benih ikan nila. 
Analysis of Htra Gene from Zebrafish (Danio Rerio) M. Murwantoko; Chio Oka; Masashi Kawaichi
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (186.97 KB) | DOI: 10.22146/ijbiotech.7554

Abstract

HtrA which is characterized by the combination of a trypsin-like catalytic domain with at least one C-terminal PDZ domain is a highly conserved family of serine proteases found in a wide range of organisms. However the identified HtrA family numbers varies among spesies, for example the number of mammalian, Eschericia coli, fruit fly-HtrA family are 4, 3 and 1 gene respectively. One gene is predicted exist in zebrafish. Since no complete information available on zebrafish HtrA, in this paper zebrafish HtrA (zHtrA) gene was analyzed. The zHtrA is belonged to HtrA1 member and predicted encodes 478 amino acids with a signal peptide, a IGF binding domain, a Kazal-type inhibitor domain in the up stream of HtrA-bacterial homolog. At the amino acid sequence the zHtrA1 showed the 69%, 69%, 68%, 54% and 54% with the rat HtrA1, mouse HtrA1, human HtrA1, human HtrA3 and mouse HtrA4 respectively. The zHtrA1 is firstly expressed at 60 hpf and mainly in the vertebral rudiments in the tail region.
Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate M. Murwantoko; Jaka Widada; Yani Lestari Nuraini
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (646.307 KB) | DOI: 10.22146/ijbiotech.7795

Abstract

Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during active penetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targeted cell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasite successfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorus vacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently, this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone and sequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique. Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA was used as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor from Riboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinant plasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agar containing X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in the LB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order to identify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolated using alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid was cut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward and M13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretory and secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the cloned gene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.', 'string'),(99, 'en_US', 'subject', 'Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2
Cloning and Sequence Analysis of Capsid Protein Gene of Iridovirus Indonesian Isolates M. Murwantoko; Christina Retna Handayani; Rarastoeti Pratiwi
Indonesian Journal of Biotechnology Vol 14, No 1 (2009)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (131.285 KB) | DOI: 10.22146/ijbiotech.7805

Abstract

Iridovirus was known as agents that caused serious systemic disease in freshwater and marine fishes. The mortality up to 100% of orange-spotted grouper (Epinephelus coioides) due to iridovirus infection has been reported in Indonesia. The gene encoding capsid protein of iridovirus is supposed to be conserved and has the potency for the development of control methods. The objectives of this study are to clone the gene encoding capsid protein iridovirus and to analyze their sequences. The   spleen tissues of orange-spotted grouper were collected and extracted their DNA. The DNA fragment of capsid protein of iridovirus genes were amplified by PCR using designed primers with the extraction DNA as templates. The amplified DNA fragments were cloned in pBSKSII and sequenced.  The genes encoding capsid protein of iridovirus from Jepara and Bali were successfully amplified and cloned. The Jepara clone (IJP03) contained complete open reading frame (ORF) of the gene composed by 1362 bp nucleotides which encoded 453 amino acids. Those Jepara and Bali (IGD01) clones shared 99.8% similarity in nucleotide level and 99.4% at amino acid level. Based on those sequences, Indonesian iridovirus was belonged to genus Megalocystivirus and shared 99,6-99,9% similarity on nucleotide level with DGIV, ISKNV, MCIV, and ALIV
Cloning and Expression of ORF124 Koi Herpesvirus as a Vaccine M. Murwantoko; Dewi Nur'aeni Setyowati; Rarastoeti Pratiwi; Masashi Kawaichi
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (226.702 KB) | DOI: 10.22146/ijbiotech.7850

Abstract

Koi herpesvirus (KHV) which also known as Cyprinid herpesvirus 3 (CyHV-3), Koi herpes-like virus,and carp interstitial nephritis gill necrosis virus (CNGV), caused signifi cant morbidity and mortality in koiand common carp (Cyprinus carpio). The case fatality rate of this disease is 80–100%. Glycoprotein has beenused for vaccine development as sub unit vaccine against viruses. The aim of this research was to clone andexpress membrane glycoprotein ORF124 KHV as a candidate of recombinant vaccine. ORF124 KHV gene wassuccessfully cloned into pBSKS and sequenced. Result showed that ORF124 KHV (isolate from Indonesia) had100 % similarity with Cyprinid herpesvirus 3 strain TUMST1 (from Japan), 99% similarity with Koi herpesvirus strain KHV-U (from USA) and Koi herpesvirus strain KHV-I (from Israel). Prediction analysis of T and B cellepitopes showed that ORF124 KHV protein had 14 and 11 T cell epitopes (IAd, Rothbard/Taylor pattern),and had 10 B cell epitopes, suggested that the protein can be used as a vaccine candidate. ORF124 gene hasbeen expressed in Escherichia coli under pET32-a(+)vector.
Limited evidence for white spot syndrome virus susceptibility associated with expression of PmVRP15 in local population of giant tiger shrimp (Penaeus monodon) Aushia Tanzih Al Haq; M. Murwantoko; T. Trijoko; Nastiti Wijayanti; Ch. Retna Handayani; Rarastoeti Pratiwi
Indonesian Journal of Biotechnology Vol 20, No 2 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (912.481 KB) | DOI: 10.22146/ijbiotech.24189

Abstract

White spot syndrome virus (WSSV) is a devastating viral disease in shrimp aquaculture. Infection ofWSSV in penaeid shrimps affects immune defense and changes gene expression. PmVRP15 has been reported as a part of the WSSV propagation pathway that is highly up-regulated in hemocytes at the acute phase of WSSV infection. This study analyzed the expression of PmVRP15 in local populations of giant tiger shrimp (Penaeus monodon) to be associated with susceptibility to WSSV. Tested populations consisted of an inbreeding population (G8) and outbreeding population (G8iA) from Jepara, Indonesia. Susceptibility was determined by cumulative mortality, median lethal time (LT50), and severity of infection at time of death. Though all populations were susceptible to WSSV, the frst mortality in G8 occurred at 18 hours post-infection (hpi) with mild infection, while frst mortality of G8iA occurred at 30 hpi with severe infection. The LT50 of G8 was signifcantly lower than that of G8iA, indicating that G8iA was less susceptible to WSSV than G8. Relative PmVRP15 transcripts of G8iA were insignifcantly down-regulated, whereas relative PmVRP15 transcripts of G8were insignifcantly upregulated. Although it’s still not conclusive, the results of this study suggest that PmVRP15 has weak potentialas a WSSV susceptibility marker in G8 and G8iA broodstock selection.
Physiological, biochemical and HSP70 and HSP90 gene expression profiles of tropical abalone Haliotis squamata in response to Vibrio alginolyticus infection Ngurah S. Yasa; Murwantoko Murwantoko; Niken S. N. Handayani; Gemi Triastutik; Lutfi Anshory
Indonesian Journal of Biotechnology Vol 25, No 1 (2020)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.51322

Abstract

Vibrio spp. have been known responsible for fish diseases in marine and brackish‐water systems in the tropics regions. Heat shock proteins are a highly conserved protein group that is known for its rapid response to environmental stresses, including infection. This study aimed to investigate physiological and biochemical responses of tropical abalone Haliotis squamata to Vibrio alginolyticus infection. Abalones were infected with V. alginolyticus by intramuscular injection at a dose of 105,106,107 cfu/abalone. The expression of HSP70 and HSP90 genes, the activity of superoxide dismutase, phenol oxidase and catalase enzymes, histology, falling and mortality were observed at 12, 24, 48, 72, and 96 hours post‐infection (hpi). The different expression of HSPs was found in this study. While the expression of HSP70 was downregulated after infection, the expression of HSP90 was upregulated at 12 hpi and followed by downregulated after 24 hpi for 106 cfu infection, but expressed at a normal level for 105 infection treatment. The expression ofsuperoxide dismutase and catalase increased within 12 hpi, and the expression of phenol oxidase increased after 24 hpi. V. alginolyticus is virulent with LD50 of less than 105 cfu on H. squamata with an average weight of 5.13 g, and caused enlargement of hemolymph sinus and development intraepithelial and intramuscular abscesses.
Study of The Effects of Carboxymethyl Chitosan on The Non-specific Defense System in The Carp (Cyprinus Carpio) Ristyana Dewi Hernawati; Triyanto .; Murwantoko .
Jurnal Sain Veteriner Vol 31, No 1 (2013): JULI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1787.601 KB) | DOI: 10.22146/jsv.3502

Abstract

Abstract Carp (Cyprinus carpio) is a freshwater fish with a high economic value, but very susceptible to diseases. One of effort to increase the productivity is by enhancing non-specific defense system. The purpose of this study is to determine the effect of carboxymethyl chitosan on enhancement non-specific defense system of carp. Carboxymethyl chitosan was obtained by alkylation process in which monochloroacetic acid in alkaline conditions was added. Carboxymethyl chitosan was given to carps at dosages of 30 μg/g, 75 μg/g and 105 μg/g, by intra muscular injection respectively. Seven and 14 days after administration of carboxymethyl chitosan, measurements of non-specific immune system parameters were done. The results showed that, administration of carboxymethyl chitosan on carps affected the phagocytic activity and lymphocytes counts. However, carboxymethyl chitosan did not give any effect to NBT activity, hematocrit, number of erythtocytes and leukocytes, monocytes and neutrophil counts in blood as well.   
Pengaruh Paparan Chlorine terhadap Stress Fisiologis dan Ekspresi Gen Hsp70 dan Hsp90 pada Abalon (Haliotis squamata) Ngurah Sedana Yasa; Lutfi Anshory; Niken S.N. Handayani; Alim Isnansetyo; Murwantoko Murwantoko
Buletin Oseanografi Marina Vol 10, No 3 (2021): Buletin Oseanografi Marina
Publisher : Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/buloma.v10i3.35251

Abstract

Abalon merupakan salah satu moluska bercangkang tunggal yang memiliki nilai ekonomis tinggi dan merupakan komoditas potensial dalam peningkatan devisa Negara. Namun permasalahannya adalah mudahnya abalone mengalami stress akibat perubahan berbagai faktor lingkungan seperti suhu, salinitas, bakteri Vibrio dan bahan desinfektan seperti chlorine. Penelitian ini dilakukan untuk mengetahui tingkat stress benih abalone terhadap paparan chlorine pada gen heat shock protein (HSP) dan mengetahui perubahan enzim-enzim  antioksidan seperti SOD,CAT,PO dan perubahan struktur histologi otot kaki abalone akibat paparan chlorine. Koleksi benih abalone dengan ukuran cangkang 3-4 cm dari unit hatchery abalone, BPIU2K Karangasem Bali. Uji paparan abalone pada akuarium kaca volume 100 L dengan konsentrasi chlorine 10 ppm. Pengambilan sample (hemolim, otot kaki, gonad) dilakukan pada waktu pengamatan (0,12,24,48 jam). Pengamatan meliputi uji  ekspresi gen heat shock protein (Hsp70 dan Hsp90), aktifitas enzim-enzim antioksidan dan histology pada otot kaki. Hasil penelitian menunjukkan bahwa Hsp70 terekspresi paling tinggi pada hemolim abalone yaitu sebesar 350 kali lipat pada paparan jam ke 12 dibandingkan kontrol (P<0.05). Sedangkan, Hsp90 pada waktu yang sama menunjukkan tingkat stress abalone paling tinggi pada otot kaki dengan tingkat ekspresi sebesar 7 kali lipat jika dibandingkan kontrol (P<0.05).  Gen heat shock protein diekspresikan cukup tinggi pada uji paparan chlorine, namun demikian  Hsp70 menunjukkan tingkat ekspresi yang lebih tinggi jika dibanding dengan Hsp90. Hsp70 lebih sensitif sebagai marka stress abalone akibat paparan chlorine. Perubahan struktur histologi menunjukkan cemaran chlorine dapat meningkatkan ukuran diameter hemolim sinus dan kerusakan pada lapisan epithel otot kaki abalone. Abalone is one of the single-shelled mollusks which has high economic value and is a commodity in increasing the country's foreign exchange. However, the problem is that it is easy for abalone to experience stress due to the influence of various environmental factors such as temperature, salinity, Vibrio bacteria and disinfectants such as chlorine. The study was conducted to determine the stress level of abalone seeds produced by hatcheries against residual chlorine. The aim of the study were to see the stress level based on the heat shock protein (HSP) gene and to see changes in antioxidant enzymes such as SOD, CAT, PO and histological structure of abalone foot muscles due to chlorine contamination. Collection of abalone seeds with a 3-4 cm shell size from the abalone hatchery unit, BPIU2K Karangasem Bali. Abalone exposure test using a glass volume of 100 L with a chlorine concentration of 10 ppm. Furthermore, sampling was carried out (hemolime, leg muscles, gonads) at the time of observation (0.12,24,48 hours). Observations included heat shock protein gene expression (Hsp70 and Hsp90) and histology in foot muscles. The results showed that Hsp70 was the highest expressed in hemolime abalone 350 times at 12 hours exposure compared to controls (P <0.05). Meanwhile, Hsp90 at the same time showed the highest level of stress on leg muscles with an expression level of  7 times when compared to controls (P <0.05). It was concluded that the heat shock protein gene was expressed high enough in the chlorine exposure test, however, Hsp70 was more sensitive as a sign of abalone stress as indicated by a higher expression level when compared to Hsp90. Changes in the histological structure show that chlorine contamination can increase the diameter of the sinus hemolime and damage to the epithelial layer of the abalone foot muscles.
Isolation and Identification of Emestrin from Emericella nidulans and Investigation of Its Anticancer Properties MUHAMMAD NURSID; EKOWATI CHASANAH; . MURWANTOKO; SUBAGUS WAHYUONO
Microbiology Indonesia Vol. 5 No. 4 (2011): December 2011
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1271.7 KB) | DOI: 10.5454/mi.5.4.3

Abstract

The research to isolate, identify and investigate of anticancer properties of active compound produced by Emericella nidulans marine fungus has been done. Active compound was isolated from mycelium extract of the  fungus. The molecular formula of active compound was established as C27H21N2O10S2 by LC-ESI-ToF-MS (m/z 597.1105 [M – H]-. Elucidation of molecular structure using FT-IR, LC-ESI-ToF-MS, 1H-NMR, 13C-NMR, and DEPT 135˚ showed that the active compound was emestrin. Emestrin was found to have cytotoxic effect against T47D, HepG2, C28, and HeLa but it was not too toxic against Vero cells with IC50 value of 1.8 µg mL-1, 4.2 µg mL-1, 2.6 µg mL-1, 13.8 µg mL-1, and 260.9 μg mL-1, respectively. Base on the cell cycle analysis by using flow cytometry, emestrin treatment at concentration of 1.0 μg mL-1 induced cell-cycle arrest in G0/G1 phase whereas at concentration of 3.0 μg mL-1, a sub-population of cells (sub G1) appeared. The apoptosis assay by using Annexin-V-FLUOS revealed that most of T47D cell treated with the compound at 1.0 and 3.0 μg mL-1 underwent apoptosis (83.6% and 92.6%, respectively. This anticancer activity of emestrin may be related to the unique of the epithiodioxopiperazine moiety with internal disulphide bond of this compound.