In experimental challenge with infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), MrNV alone can cause mortality in freshwater prawn (Macrobrachium rosenbergii)

Warachin Gangnonngiw, Malinee Bunnontae, Kornsunee Phiwsaiya, Saengchan Senapin, Arun K. Dhar

Research output: Contribution to journalArticle

Abstract

To overcome the lack of immortal shrimp cell lines for shrimp viral research, we constructed and tested DNA infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) often found together in freshwater prawn (M. rosenbergii) exhibiting white tail disease (WTD). Full-length cDNAs of MrNV and XSV genomic RNA were individually inserted into the baculovirus pFastBacDUAL shuttle vector. Individual Sf9 (insect cell line) transfection resulted in production of RNA (RT-PCR) and capsid proteins (immunofluorescence) for both viruses. Presence of respective virions was confirmed by density gradient purification followed by RT-PCR and transmission electron microscopy. Infectivity was by tested in immersion-challenge tests with M. rosenbergii post-larvae (PL) using both semi-purified viruses, individually or combined, and confirmed by histological analysis (morphology and immunofluorescence) and quantitative RT-PCR. Mortality accompanied by WTD lesions occurred with MrNV alone or in combination with XSV but not with XSV alone, despite its replication.

Original languageEnglish (US)
Pages (from-to)30-37
Number of pages8
JournalVirology
Volume540
DOIs
StatePublished - Jan 15 2020

Fingerprint

Palaemonidae
Fresh Water
Clone Cells
RNA
Viruses
Mortality
Fluorescent Antibody Technique
Tail
Sf9 Cells
Cell Line
Genetic Vectors
Baculoviridae
Capsid Proteins
Immersion
Transmission Electron Microscopy
Virion
Transfection
Larva
Insects
Complementary DNA

Keywords

  • Freshwater prawn
  • Infectious clone
  • M. rosenbergii
  • MrNV
  • White tail disease
  • XSV

ASJC Scopus subject areas

  • Virology

Cite this

In experimental challenge with infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), MrNV alone can cause mortality in freshwater prawn (Macrobrachium rosenbergii). / Gangnonngiw, Warachin; Bunnontae, Malinee; Phiwsaiya, Kornsunee; Senapin, Saengchan; Dhar, Arun K.

In: Virology, Vol. 540, 15.01.2020, p. 30-37.

Research output: Contribution to journalArticle

@article{88753654672a4f75ad5ac520a9fea4d6,
title = "In experimental challenge with infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), MrNV alone can cause mortality in freshwater prawn (Macrobrachium rosenbergii)",
abstract = "To overcome the lack of immortal shrimp cell lines for shrimp viral research, we constructed and tested DNA infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) often found together in freshwater prawn (M. rosenbergii) exhibiting white tail disease (WTD). Full-length cDNAs of MrNV and XSV genomic RNA were individually inserted into the baculovirus pFastBacDUAL shuttle vector. Individual Sf9 (insect cell line) transfection resulted in production of RNA (RT-PCR) and capsid proteins (immunofluorescence) for both viruses. Presence of respective virions was confirmed by density gradient purification followed by RT-PCR and transmission electron microscopy. Infectivity was by tested in immersion-challenge tests with M. rosenbergii post-larvae (PL) using both semi-purified viruses, individually or combined, and confirmed by histological analysis (morphology and immunofluorescence) and quantitative RT-PCR. Mortality accompanied by WTD lesions occurred with MrNV alone or in combination with XSV but not with XSV alone, despite its replication.",
keywords = "Freshwater prawn, Infectious clone, M. rosenbergii, MrNV, White tail disease, XSV",
author = "Warachin Gangnonngiw and Malinee Bunnontae and Kornsunee Phiwsaiya and Saengchan Senapin and Dhar, {Arun K.}",
year = "2020",
month = "1",
day = "15",
doi = "10.1016/j.virol.2019.11.004",
language = "English (US)",
volume = "540",
pages = "30--37",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - In experimental challenge with infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), MrNV alone can cause mortality in freshwater prawn (Macrobrachium rosenbergii)

AU - Gangnonngiw, Warachin

AU - Bunnontae, Malinee

AU - Phiwsaiya, Kornsunee

AU - Senapin, Saengchan

AU - Dhar, Arun K.

PY - 2020/1/15

Y1 - 2020/1/15

N2 - To overcome the lack of immortal shrimp cell lines for shrimp viral research, we constructed and tested DNA infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) often found together in freshwater prawn (M. rosenbergii) exhibiting white tail disease (WTD). Full-length cDNAs of MrNV and XSV genomic RNA were individually inserted into the baculovirus pFastBacDUAL shuttle vector. Individual Sf9 (insect cell line) transfection resulted in production of RNA (RT-PCR) and capsid proteins (immunofluorescence) for both viruses. Presence of respective virions was confirmed by density gradient purification followed by RT-PCR and transmission electron microscopy. Infectivity was by tested in immersion-challenge tests with M. rosenbergii post-larvae (PL) using both semi-purified viruses, individually or combined, and confirmed by histological analysis (morphology and immunofluorescence) and quantitative RT-PCR. Mortality accompanied by WTD lesions occurred with MrNV alone or in combination with XSV but not with XSV alone, despite its replication.

AB - To overcome the lack of immortal shrimp cell lines for shrimp viral research, we constructed and tested DNA infectious clones of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) often found together in freshwater prawn (M. rosenbergii) exhibiting white tail disease (WTD). Full-length cDNAs of MrNV and XSV genomic RNA were individually inserted into the baculovirus pFastBacDUAL shuttle vector. Individual Sf9 (insect cell line) transfection resulted in production of RNA (RT-PCR) and capsid proteins (immunofluorescence) for both viruses. Presence of respective virions was confirmed by density gradient purification followed by RT-PCR and transmission electron microscopy. Infectivity was by tested in immersion-challenge tests with M. rosenbergii post-larvae (PL) using both semi-purified viruses, individually or combined, and confirmed by histological analysis (morphology and immunofluorescence) and quantitative RT-PCR. Mortality accompanied by WTD lesions occurred with MrNV alone or in combination with XSV but not with XSV alone, despite its replication.

KW - Freshwater prawn

KW - Infectious clone

KW - M. rosenbergii

KW - MrNV

KW - White tail disease

KW - XSV

UR - http://www.scopus.com/inward/record.url?scp=85074791115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85074791115&partnerID=8YFLogxK

U2 - 10.1016/j.virol.2019.11.004

DO - 10.1016/j.virol.2019.11.004

M3 - Article

C2 - 31734381

AN - SCOPUS:85074791115

VL - 540

SP - 30

EP - 37

JO - Virology

JF - Virology

SN - 0042-6822

ER -