Characterization of a novel picornavirus isolated from moribund aquacultured clownfish | Microbiology Society

Characterization of a novel picornavirus isolated from moribund aquacultured clownfish | Microbiology Society
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1887
Journal of General Virology
Volume 101, Issue 7
Research Article
Characterization of a novel picornavirus isolated from moribund aquacultured clownfish
Elizabeth C. Scherbatskoy
1​
Kuttichantran Subramaniam
1​
Lowia Al-Hussinee
1​
Kamonchai Imnoi
1​
Patrick M. Thompson
1​,†​
Vsevolod L. Popov
2​
Terry Fei Fan Ng
3​
Karen L. Kelley
4​
Rodolfo Alvarado
4​
Jeffrey C. Wolf
5​
Deborah B. Pouder
6​
Roy P.E. Yanong
6​
and
Thomas B. Waltzek
1​
View Affiliations
Hide Affiliations
1​
Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA
2​
Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA
3​
College of Veterinary Medicine, University of Georgia, Athens, GA, USA
4​
Electron Microscopy Core, Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, FL, USA
5​
Experimental Pathology Laboratories, Inc., Sterling, VA, USA
6​
Tropical Aquaculture Laboratory, Program in Fisheries and Aquatic Sciences, School of Forest Resources and Conservation, IFAS, University of Florida, Ruskin, FL, USA
†​
Present address: Whitney Laboratory for Marine Bioscience, St Augustine, FL, USA
*Correspondence:
Thomas B. Waltzek,
[email protected]
Published:
18 May 2020
Abstract
Over the last decade, a number of USA aquaculture facilities have experienced periodic mortality events of unknown aetiology in their clownfish (
Amphiprion ocellaris
). Clinical signs of affected individuals included lethargy, altered body coloration, reduced body condition, tachypnea, and abnormal positioning in the water column. Samples from outbreaks were processed for routine parasitological, bacteriological, and virological diagnostic testing, but no consistent parasitic or bacterial infections were observed. Histopathological evaluation revealed individual cell necrosis and mononuclear cell inflammation in the branchial cavity, pharynx, oesophagus and/or stomach of four examined clownfish, and large basophilic inclusions within the pharyngeal mucosal epithelium of one fish. Homogenates from pooled external and internal tissues from these outbreaks were inoculated onto striped snakehead (SSN-1) cells for virus isolation and cytopathic effects were observed, resulting in monolayer lysis in the initial inoculation and upon repassage. Transmission electron microscopy of infected SSN-1 cells revealed small round particles (mean diameter=20.0–21.7 nm) within the cytoplasm, consistent with the ultrastructure of a picornavirus. Full-genome sequencing of the purified virus revealed a novel picornavirus most closely related to the bluegill picornavirus and other members of the genus
Limnipivirus
. Additionally, pairwise protein alignments between the clownfish picornavirus (CFPV) and other known members of the genus
Limnipivirus
yielded results in accordance with the current International Committee on Taxonomy of Viruses criteria for members of the same genus. Thus, CFPV represents a proposed new limnipivirus species. Future experimental challenge studies are needed to determine the role of CFPV in disease.
Received:
24/02/2020
Accepted:
06/04/2020
Published Online:
18/05/2020
Keyword(s):
cell culture
clownfish
emerging disease
molecular diagnostics
NGS
ornamental aquaculture
PCR
picornavirus
and
viral discovery
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References
Zell
Delwart
Gorbalenya
AE
Hovi
King
AMQ
et al.
ICTV Virus Taxonomy Profile:
Picornaviridae
J Gen Virol
2017
; 98:
2421–
2422
[View Article]
[PubMed]
[Google Scholar]
King
AMQ
Lefkowitz
EJ
Mushegian
AR
Adams
MJ
Dutilh
BE
et al.
Changes to taxonomy and the International Code of virus classification and nomenclature ratified by the International Committee on taxonomy of viruses (2018).
Arch Virol
2018
; 163:
2601–
2631
[View Article]
[PubMed]
[Google Scholar]
Racaniello
VP
Picornaviridae
: The viruses and their replication.
In
Knipe
DM
Howley
PM
(editors)
Fields Virology
, 6th ed. Philadelphia, PA: Lippincott Williams & Wilkins;
2013
pp
453–
489
[Google Scholar]
Jiang
Liu
Ma
H-C
Paul
AV
Wimmer
Picornavirus morphogenesis.
Microbiol Mol Biol Rev
2014
; 78:
418–
437
[View Article]
[PubMed]
[Google Scholar]
Woo
PCY
Lau
SKP
Choi
GKY
Huang
Teng
JLL
et al.
Natural occurrence and characterization of two internal ribosome entry site elements in a novel virus, canine picodicistrovirus, in the picornavirus-like superfamily.
J Virol
2012
; 86:
2797–
2808
[View Article]
[PubMed]
[Google Scholar]
Reuter
Boros
Ákos
Földvári
Szekeres
Mátics
et al.
Dicipivirus (family
Picornaviridae
) in wild Northern white-breasted hedgehog (
Erinaceus roumanicus
).
Arch Virol
2018
; 163:
175–
181
[View Article]
[PubMed]
[Google Scholar]
Lin
J-Y
Chen
T-C
Weng
K-F
Chang
S-C
Chen
L-L
et al.
Viral and host proteins involved in picornavirus life cycle.
J Biomed Sci
2009
; 16:
103
[View Article]
[PubMed]
[Google Scholar]
TFF
Personal observation;
2019
Moore
AR
LI
MF
McMENEMY
Isolation of a picorna-like virus from smelt,
Osmerus mordax
(Mitchill).
J Fish Dis
1988
; 11:
179–
184
[View Article]
[Google Scholar]
Ahne
Anders
Halder
Yoshimizu
Isolation of picornavirus-like particles from the European smelt,
Osmerus eperlanus
(L.).
J Fish Dis
1990
; 13:
167–
168
[View Article]
[Google Scholar]
Hedrick
RP
McDowell
TS
Kent
ML
Elston
RA
A small RNA virus isolated from Atlantic salmon (
Salmo salar
).
J Appl Ichthyol
1990
; 6:
173–
181
[View Article]
[Google Scholar]
Hedrick
RP
Yun
Wingfield
WH
A small RNA virus isolated from salmonid fishes in California, USA.
Can J Fish Aquat Sci
1991
; 48:
99–
104
[View Article]
[Google Scholar]
Eaton
WD
Bagshaw
Hulett
Evans
Isolation of a picorna-like virus from steelhead in Washington state.
J Aquat Anim Health
1992
; 4:
90–
96
[View Article]
[Google Scholar]
Glazebrook
JS
Heasman
MP
BEER
SW
Picorna-like viral particles associated with mass mortalities in larval barramundi,
Lates calcarifer
Bloch.
J Fish Dis
1990
; 13:
245–
249
[View Article]
[Google Scholar]
Munday
BL
Langdon
JS
Hyatt
Humphrey
JD
Mass mortality associated with a viral-induced vacuolating encephalopathy and retinopathy of larval and juvenile barramundi,
Lates calcarifer
Bloch.
Aquaculture
1992
; 103:
197–
211
[View Article]
[Google Scholar]
Bloch
Gravningen
Larsen
JL
Encephalomyelitis among turbot associated with a picornavirus-like agent.
Dis Aquat Organ
1991
; 10:
65–
70
[View Article]
[Google Scholar]
Breuil
Bonami
JR
Pepin
JF
Pichot
Viral infection (picorna-like virus) associated with mass mortalities in hatchery-reared sea-bass (
Dicentrarchus labrax
) larvae and juveniles.
Aquaculture
1991
; 97:
109–
116
[View Article]
[Google Scholar]
Mori
K-ichiro
Nakai
Nagahara
Muroga
Mekuchi
et al.
A viral disease in hatchery-reared larvae and juveniles of redspotted grouper.
Fish Pathol
1991
; 26:
209–
210
[View Article]
[Google Scholar]
Iwanowicz
LR
Goodwin
AE
Heil
A small RNA virus isolated from apparently healthy wild sandbar shiners,
Notropis scepticus
(Jordan & Gilbert).
J Fish Dis
2000
; 23:
349–
352
[View Article]
[Google Scholar]
Petty
BD
Fraser
WA
Viruses of PET fish.
Vet Clin North Am Exot Anim Pract
2005
; 8:
67–
84
[View Article]
[PubMed]
[Google Scholar]
Reuter
Pankovics
Delwart
Boros
Ákos
A novel posavirus-related single-stranded RNA virus from fish (
Cyprinus carpio
).
Arch Virol
2015
; 160:
565–
568
[View Article]
[PubMed]
[Google Scholar]
Batts
Yun
Hedrick
Winton
A novel member of the family Hepeviridae from cutthroat trout (Oncorhynchus clarkii).
Virus Res
2011
; 158:
116–
123
[View Article]
[PubMed]
[Google Scholar]
Bovo
Florio
Viral diseases of cultured marine fish.
In
Eiras
JC
Segner
Wahli
Kapoor
BG
(editors)
Fish Diseases
, 1st ed. Science Publishers: Enfield, NH;
2008
pp
185–
238
[Google Scholar]
Walker
PJ
Winton
JR
Emerging viral diseases of fish and shrimp.
Vet Res
2010
; 41:
51
[View Article]
[PubMed]
[Google Scholar]
Barbknecht
Sepsenwol
Leis
Tuttle-Lau
Gaikowski
et al.
Characterization of a new picornavirus isolated from the freshwater fish
Lepomis macrochirus
J Gen Virol
2014
; 95:
601–
613
[View Article]
[PubMed]
[Google Scholar]
Lange
Groth
Fichtner
Granzow
Keller
et al.
Virus isolate from carp: genetic characterization reveals a novel picornavirus with two aphthovirus 2A-like sequences.
J Gen Virol
2014
; 95:
80–
90
[View Article]
[PubMed]
[Google Scholar]
Phelps
NBD
Mor
SK
Armien
AG
Batts
Goodwin
AE
et al.
Isolation and molecular characterization of a novel picornavirus from baitfish in the USA.
PLoS One
2014
; 9:
e87593
[View Article]
[PubMed]
[Google Scholar]
Fichtner
Philipps
Groth
Schmidt-Posthaus
Granzow
et al.
Characterization of a novel picornavirus isolate from a diseased European eel (
Anguilla anguilla
).
J Virol
2013
; 87:
10895–
10899
[View Article]
[PubMed]
[Google Scholar]
Hahn
MA
Dheilly
NM
Genome characterization, prevalence, and transmission mode of a novel picornavirus associated with the threespine stickleback fish (
Gasterosteus aculeatus
).
J Virol
2019
; 93:
e02277–
18
[View Article]
[PubMed]
[Google Scholar]
Altan
Kubiski
SV
Boros
Ákos
Reuter
Sadeghi
et al.
A highly divergent picornavirus infecting the gut epithelia of zebrafish (
Danio rerio
) in research institutions world-wide.
Zebrafish
2019
; 16:
291–
299
[View Article]
[PubMed]
[Google Scholar]
Geoghegan
JL
Di Giallonardo
Cousins
Shi
Williamson
JE
et al.
Hidden diversity and evolution of viruses in market fish.
Virus Evol
2018
; 4:
vey031
[View Article]
[PubMed]
[Google Scholar]
Shi
Lin
X-D
Chen
Tian
J-H
Chen
L-J
et al.
The evolutionary history of vertebrate RNA viruses.
Nature
2018
; 556:
197–
202
[View Article]
[PubMed]
[Google Scholar]
Fautin
DG
Allen
GR
Field Guide to Anemone Fishes and Their Host Sea Anemones
Perth: Western Australian Museum;
1992
[Google Scholar]
Sin
TM
Teo
MM
Ng
PKL
Chou
LM
Khoo
HW
The damselfishes (Pisces: Osteichthyes: Pomacentridae) of Peninsular Malaysia and Singapore: systematics, ecology and conservation.
Hydrobiologia
1994
; 285:
49–
58
[View Article]
[Google Scholar]
Patkaew
Direkbusarakom
Tantithakura
A simple method for cell culture of ‘Nemo’ ocellaris clownfish (
Amphiprion ocellaris
, Cuvier 1830).
Cell Bio Int Rep
2014
; 21:
39–
45
[Google Scholar]
King
TA
Wild caught ornamental fish: a perspective from the UK ornamental aquatic industry on the sustainability of aquatic organisms and livelihoods.
J Fish Biol
2019
; 94:
925–
936
[View Article]
[PubMed]
[Google Scholar]
Watson
CA
Shireman
JV
Production of Ornamental Aquarium Fish
University of Florida IFAS Extension;
2002
[Google Scholar]
Wabnitz
Taylor
Green
Razak
From ocean to aquarium: a global trade in marine ornamental species. UNEP world conservation and monitoring centre (WCMC);
2003
Sirajudheen
TK
Shyam
SS
Bijukumar
Bindu
Problems and prospects of marine ornamental fish trade in Kerala, India.
J Fish Econ & Dev
2014
; 15:
14–
30
[Google Scholar]
Yanong
Personal observation;
2018
Ahasan
MS
Subramaniam
Sayler
KA
Loeb
JC
Popov
VL
et al.
Molecular characterization of a novel reassortment
mammalian orthoreovirus
type 2 isolated from a Florida white-tailed deer fawn.
Virus Res
2019
; 270:
197642
[View Article]
[Google Scholar]
Schindelin
Rueden
CT
Hiner
MC
Eliceiri
KW
The ImageJ ecosystem: an open platform for biomedical image analysis.
Mol Reprod Dev
2015
; 82:
518–
529
[View Article]
[PubMed]
[Google Scholar]
Kearse
Moir
Wilson
Stones-Havas
Cheung
et al.
Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data.
Bioinformatics
2012
; 28:
1647–
1649
[View Article]
[PubMed]
[Google Scholar]
Nguyen
L-T
Schmidt
HA
von Haeseler
Minh
BQ
IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies.
Mol Biol Evol
2015
; 32:
268–
274
[View Article]
[PubMed]
[Google Scholar]
Muhire
BM
Varsani
Martin
DP
SDT: a virus classification tool based on pairwise sequence alignment and identity calculation.
PLoS One
2014
; 9:
e108277
[View Article]
[PubMed]
[Google Scholar]
Rivers
TM
Viruses and Koch's postulates.
J Bacteriol
1937
; 33:
1–
12
[View Article]
[PubMed]
[Google Scholar]
Dee
LE
Horii
SS
Thornhill
DJ
Conservation and management of ornamental coral reef wildlife: successes, shortcomings, and future directions.
Biol Conserv
2014
; 169:
225–
237
[View Article]
[Google Scholar]
/content/journal/jgv/10.1099/jgv.0.001421
Characterization of a novel picornavirus isolated from moribund aquacultured clownfish
J. Gen. Virol.
101
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(2020)
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Characterization of a novel picornavirus isolated from moribund aquacultured clownfish
Publication Date:
18 May 2020