Delivery of double stranded RNA by Macrobrachium rosenbergii nodavirus-like particles to protect shrimp from white spot syndrome virus

Jariyapong, Pitchanee; Chotwiwatthanakun, Charoonroj; Direkbusarakom, Sataporn; Hirono, Ikuo; Wuthisuthimethavee, Suwit; Weerachatyanukul, Wattana

doi:10.1016/j.aquaculture.2014.09.034

Cited by 41 publications

(37 citation statements)

References 35 publications

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“…Nilai RPS pada penelitian ini relatif sama dengan hasil penelitian Witteveldt et al (2004b) pada udang windu dengan menggunakan vaksin VP-28 dan campuran VP-28+VP-19 dengan nilai masing-masing adalah 44% dan 33% pada pengamatan uji tantang pada hari ke-2 setelah vaksinasi. Namikoshi et al (2004) melaporkan bahwa aplikasi vaksin VP-26 dan VP-28 pada udang P. japonicus dapat meningkatkan resistensinya dengan nilai RPS masing-masing 80% dan 95%, sedangkan aplikasi dsRNA VP-28 pada Macrobrachium rosenbergii menghasilkan RPS sebesar 44,5% terhadap WSSV (Jariyapong et al, 2015). Vaksinasi melalui pakan pada udang windu dengan menggunakan VP-19 dapat meningkatan sintasan dengan nilai RPS sebesar 77% pada hari ke-7 dan 29% pada hari ke-21 setelah uji tantang dengan WSSV (Witteveldt et al, 2004a).…”

Section: Aplikasi Vaksin Rekombinan Dsrna Vp-15unclassified

Pengklonan Gen Penyandi Viral Protein 15 (Vp-15) WSSV Dan Aplikasinya Sebagai Vaksin Rekombinan Pada Udang Windu

Parenrengi

Mulyaningrum

Tenriulo

et al. 2018

Jurnal Riset Akuakultur

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ABSTRAKInfeksi white spot syndrome virus (WSSV) dapat menyebabkan kematian massal pada budidaya udang windu Penaeus monodon di Indonesia. Infeksi yang terjadi secara sistematis tersebut disebabkan oleh peran gen nucleocapsid viral protein . Upaya pengembangan gen VP-15 WSSV untuk menginduksi respons imun dan menetralisasi terhadap infeksi WSSV pada udang windu perlu dilakukan. Penelitian ini bertujuan untuk mengisolasi dan merekombinasikan gen penyandi VP-15 WSSV sebagai vaksin dsRNA, serta menganalisis aplikasinya pada udang windu. Gen VP-15 diisolasi dari udang windu yang terinfeksi WSSV, dikloning ke dalam suatu vektor dan ditransformasikan ke sel kompeten (bakteri Escheria coli DH5). Plasmid diisolasi untuk mengonfirmasi insert region gen VP-15 melalui sekuensing nukleotida. Pembuatan vaksin rekombinan dilakukan secara in-vitro menggunakan kit MEGAscript RNAi dan diaplikasikan ke udang windu melalui metode injeksi dengan dosis tunggal 0,2 µg dan kontrol (tanpa injeksi vaksin). Hewan uji yang digunakan berukuran panjang 14,75±3,17 g dan bobot 11,64±0,76 cm; serta dipelihara pada wadah bak fiber volume 250 L dengan kepadatan 10 ekor/bak. Hasil penelitian menunjukkan bahwa gen penyandi VP-15 telah diisolasi dari udang windu dan vaksin rekombinan telah dihasilkan secara in-vitro. Analisis sekuens nukleotida memperlihatkan bahwa sisipan gen DNA VP-15 sebesar 253 bp dan menunjukkan kemiripan yang tinggi (99%) pada GenBank. Penggunaan vaksin rekombinan dsRNA dengan dosis 0,2 µg memperlihatkan sintasan udang windu yang dapat mencapai 40,0% dibandingkan dengan kontrol hanya 3,3% (peningkatan 36,7%). Gambaran histopatologi pada jaringan hepatopankreas udang windu pada perlakuan kontrol menunjukkan adanya kerusakan inti sel, akibat infeksi WSSV. Gene VP-15 berpotensi sebagai bahan vaksin rekombinan dsRNA dalam mencegah infeksi WSSV.

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Section: Aplikasi Vaksin Rekombinan Dsrna Vp-15unclassified

Pengklonan Gen Penyandi Viral Protein 15 (Vp-15) WSSV Dan Aplikasinya Sebagai Vaksin Rekombinan Pada Udang Windu

Parenrengi

Mulyaningrum

Tenriulo

et al. 2018

Jurnal Riset Akuakultur

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“…The application of antisense RNA in protecting L. vannamei from WSSV infection was also reported by Akhila et al (2015). The dsRNA from VP-28 was reported to be useful in protecting Macrobrachium rosenbergii from WSSV (Jariyapong et al, 2015). Several studies also demonstrated the success of VP-19 isolation from the WSSV infected shrimp (Alim et al, 2011;Hidayani et al, 2016).…”

Section: Characteristic Of a Viral Protein (Vp-15) Of White Spot Syndmentioning

confidence: 80%

CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798)

Parenrengi

Alimuddin

Tenriulo

2017

Indonesian Aquaculture Journal

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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.

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“…Despite many reports that dsRNA is ready to use without any adjuvants or carriers, our previous results demonstrate the benefit of using an MrNV virus-like particle (MrNV-VLP)-based delivery system to carry therapeutic agents for curing WSSV infection. A higher survival rate was observed in the group of shrimp receiving encapsulated VP28 dsRNA-VLP compared to other control groups (Jariyapong et al, 2015). In this study, we further investigated the levels of RNAi machinery for antiviral defense mechanism as well as immune and apoptotic related genes in shrimp receiving encapsulated VP28 dsRNA to validate a novel potential therapeutic approach for effective control of this shrimp viral infection.…”

Section: Introductionmentioning

confidence: 91%

“…RT-PCR and RT-qPCR Jariyapong et al (2015) use. Before the viral challenge, an aliquot was thawed and centrifuged (3000 ×g, 4°C, 10 min).…”

Section: Genementioning

confidence: 99%

Enhancement of shrimp immunity against white spot syndrome virus by Macrobrachium rosenbergii nodavirus-like particle encapsulated VP28 double-stranded RNA

et al. 2015

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Delivery of double stranded RNA by Macrobrachium rosenbergii nodavirus-like particles to protect shrimp from white spot syndrome virus

Cited by 41 publications

References 35 publications

Pengklonan Gen Penyandi Viral Protein 15 (Vp-15) WSSV Dan Aplikasinya Sebagai Vaksin Rekombinan Pada Udang Windu

Pengklonan Gen Penyandi Viral Protein 15 (Vp-15) WSSV Dan Aplikasinya Sebagai Vaksin Rekombinan Pada Udang Windu

CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798)

Enhancement of shrimp immunity against white spot syndrome virus by Macrobrachium rosenbergii nodavirus-like particle encapsulated VP28 double-stranded RNA

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