Aquaculture Viruses

What a Virus Isnt

Not a bacterium...
Not independent...
Cannot survive in absence of a living cell
within which to replicate...
Antibiotics generally dont work on them...

What Viruses Are...

Infectious agents composed mainly of nucleic acid
with a protein coat (capsid)
Visible with electron microscope (10-200 nM)
Carry on normal cell-like function (unless free, then
infectious)
In infectious form: no growth; no respiration???
Can enter living plant, animal or human

Virus Appearence?
Capsid, core and genetic material (DNA/RNA)
Capsid: outer shell of the virus which encloses genetic material
(link: chemical structure of capsid helps determine immune
response to virus)
capsid is made of many identical individual proteins
protein core under capsid protecting genetic material
sometimes an additional covering (lipid bilayer w/embedded
proteins) on outside known as an envelope ( like a baseball)
various forms: rods, filaments, spheres, cubes, crystals

Capsid

capsomere: unit/molecule associated with capsid structure

Typical Virus Shapes

SPHERES

RODS

CUBES

More Virus Shapes

Composition of T-Even
Bacteriophage
Capsid: brains of virus,
tightly-wound protein
protecting nucleic acids
Body: attached to
capsid head, rod-like
structure w/retractible
sheath, hollow core
Tail: at end of core is a
spiked plate carrying 6
slender tail fibers,
anchor virus to its host

How do viruses work?

Viruses make use of the host cells chemical energy,
protein and nucleic acid synthesizing ability to replicate
themselves...
each virus attacks a specific type of cell
cold viruses attack cells of the lung
the AIDS virus attacks T4 cells of the
immune system
fish viruses are just as specific

Bacteriophage Attack

Virusal Mechanism
Viruses contain single- or double- stranded DNA
or RNA
Often, the virus alters the intracellular
environment enough to damage or kill the cell
(oops!!)

If enough cells are destroyed, disease results!

Role of RNA/DNA
Supplies the codes for building the protein coat
(capsid) and for producing enzymes needed to
replicate more viruses
Information given so newly-built viruses can lyse cells
(e.g., bacteriophage)
Result: cell destroyed.

Bottom Line...
All viruses only exist to make more viruses
Most are harmful
Replication = host cell death.

The Virus Invasion: Step by step

Phase 1: Attachment of virus to bacteria, etc.
Phase 2: injection of DNA or RNA

Phase 3: DNA (RNA) enters the host cell

Phase 4:DNA incorporated (10 minutes) hundreds of
virions appear causing the cell to rupture, releasing
hundreds of small viral replicates
This is how it can replicate so quickly!

The Virus Invasion

Whats Infected by a Virus?

All living things have some susceptibility to a

particular virus
Virus is specific for the organism
Within a species, there may be a 100 or more
different viruses which can affect that species
alone
Specific: for example, a virus that only affects
one organism (humans and smallpox)
Influenza can infect humans and two animals

Different Types of Viruses

Major classifications: animal, plant, bacterial
Sub-classified by arrangement and type of nucleic
acid
Animal virus group: double-stranded DNA, singlestranded DNA, double-stranded RNA, singlestranded RNA, retrovirus
Influenza: SS-RNA

Do Viruses ever Change?

Mutations do occur.
If the mutation is harmful, the new virus particle
might no longer be functional (infectious)
However, because a given virus can generate many,
many copies, a small number of non-functional
viruses is not important
Mutation is not necessarily damaging to the virus -it can lead to a functional but new strain of virus

Defense Against Viruses

First Line: skin and mucous membrane, which
also lines the gastrointestinal and respiratory
passageways
skin is tough and stomach acidity acts as a
disinfectant
Second Line: after the virus enters the blood and
other tissues, white blood cells and related cells
(phagocytes) consume them
accumulation of phagocytes in area of infection is
known as puss

Defense Against Viruses

Antibodies attacking chickenpox virus

Defense Against Viruses

Antibodies are the best defense against viruses

unfortunately, they are specific in their action
chickenpox antibody will only attack a chickenpox virus
a particular virus stimulates the production of a particular
antibody

Defense Against Viral Infection

Animals are protected in several ways:
1) intracellular: if a particular virus attacks cells, our
bodies produce interferons
interferons (alpha, beta or gamma) are proteins which
interact with adjacent cells and cause them to become more
resistant to infection by the virus
if the resistance is not quite good enough, we become sick

Defense Against Viral Infection

2) immune system (extracellular): kills the virus
outside the cell
also kills the infected cells
virus cannot spread
eventually the virus is completely removed and we
get better
exception: HIV because it infects cells of the
immune system, itself
chemicals/drugs: acyclovir, AZT, HIV protease
inhibitor, several fish vaccines available.

Major Fish Viruses

Major Viral Infections in Fish

Infectious pancreatic necrosis (IPN)

Viral hemorrhagic septicemia (VHS)

Infectious hematopoetic necrosis (IHN)

Channel catfish virus disease (CCVD)

Koi Herpes Virs
Lymphocytis
Cacar virus
VNN (Viral Nervous Necrosis)
Spring Viraemia of Carp (SVC)

(1) Infectious Pancreatic Necrosis

(IPN)
What?: viral infection of salmonids (trout and char)
Time: Acute
Result: high mortality (fry and fingerlings)
Rare in larger fish (good thing!)
History: Isolated in Pacific NW in 1960s, wiped out brook
trout in Oregon in 1971-73
Size: Only 65 nM in diam., smallest of fish viruses

IPN: general notes

Single capsid shell, icosohedral symmetry, no envelope
Contains two segments of DS-RNA

Fairly stable and resistant to chemicals (acid, ether, etc.),

variable resistance to freezing
Remains infectious for 3 months in water (uh oh!)
Targets pancreas and hematopoietic tissues of kidney and spleen

IPN: epizootiology (disease process)

Who?: All salmonids, brook trout most susceptible,
marine fish (flounder?)
Reservoirs (where)?: carriers, once a carrier always a
carrier, virus particles shed in feces/urine
Transmission (how?): horizontal, by waters via
carriers or infected fry; vertical from adults to
progeny; experimentally by feeding infected material,
IP injection
Pathogenesis: entry via gills, digestive tract
Environmental factors: mortality reduced at lower
temps (why?); however, carriers not reduced

IPN: pathology (what do we see?)

IPN: detection, diagnosis and control

Isolation: whole fry, kidney, spleen, pyloric cecae, sex
fluids are all good sources, .i.e. check these!!!

Presumptive tests: epizootiological evidence and/or typical

PCR in infected cells
Definitive tests: serology (fluorescent antibody test (FAT))
Control: avoid virus in water, virus-free stock, destruction
of infected stock, vaccine exists now!

How Bad Can It Be??

Fish severely affected by IPNV:

Atlantic salmon* (Salmo salar)
brook trout* (Salvelinus fontinalis)
brown trout* (Salmo trutta)
danio zebrafish* (Brachydanio rerio)
rainbow trout* (Oncorhynchus mykiss)
yellowtail* (Seriola lalandi)

Other species known to be

susceptible
amago salmon (Oncorhynchus rhodurus)
Arctic char (Salvelinus alpinus)
Atlantic menhadden (Brevoortia tyrannus)
carangids (Carangidae)
chinook salmon (Oncorhynchus tshawytscha)
chum salmon (Oncorhynchus keta)
cichlids (Cichlidae)
coho salmon (Oncorhynchus kisutch)
common scallop (Pecten maximus)
cutthroat trout (Salmo clarki)
cyprinids (Cyprinidae)
Danube salmon (Salmo hucho)
drums/croakers (Sciaenidae)
eels (Anguilla spp)
grayling (Thymallus thymallus)

More

halibut (Hippoglossus stenolepis)

Jap. amberjack (Seriola quinqueradiata)
lampreys (Petromyzontyidae)
loach (Misgurnus anguillicaudatus)
masou salmon (Oncorhynchus masou)
perches (Percidae)
silversides (Atherinidae)
soles (Soleidae)
striped snakehead (Channa striatus)
summer flounder (Paralichthys dentatus)
white seabass (Moronidae)
carp (Cyprinus carpio)
redfin perch (Perca fluviatilis)
yellowfin bream (Acanthopagrus australis)

herrings/sardines (Clupidae)
lake trout (Salvelinus namaycush)
left-eye flounders (Bothidae)
loaches (Cobitidae)
Pacific salmon (Oncorhynchus spp)
pikes (Esocidae)
sockeye salmon (Oncorhynchus nerka)
Southwest European nase (C. toxostoma)
suckers (Cotostomidae)
turbot (Psetta maxima)
whitefish (Coregonidae)
goldfish (Carassius auratus)
southern flounder (P. lethostigma)

Asymptomatic carriers...
coalfish (Pollachius virens)
common carp (Cyprinus carpio)
discus fish (Symphysodon discus)
goldfish (Carrasius auratus)
heron (Ardea cinerea)
loach (Cobitidae)
minnow (Phoxinus phoxinus)
noble crayfish (Astacus astacus)
Infectious pancreatic necrosis in Atlantic salmon.
pike (Esox lucius)
Note swollen stomach and 'pop eye'
river lamprey (Lampetra fluviatalis) Source: Australian Animal Health Laboratory
shore crab (Carcinus maenas)
Spanish barbel (Barbus graellsi)
white suckers (Catostomas commersoni)
...what now???

(2) Viral Hemorrhagic Septicemia (VHS)

(2) Viral Hemorrhagic Septicemia (VHS)

What?: Viral disease of European salmonids
When?: Recognized in Denmark in 1949, isolated
from Pacific Coast in 1989
Size: rhabdovirus, bullet-shaped (one rounded
end), 185 x 65 nM, lipoprotein envelope
non-segmented SS-RNA
Constitution: sensitive to ether and chloroform,
heat, acid, resistant to freeze-drying

Viral Hemorrhagic Septicemia

Produces a general viremia, tissue and organ damage, liver
necrosis, spleen, kidney
Epizootiology: cultured rainbow trout, also brown trout,
steelhead, chinook, coho (most cases in WA state)
Reservoirs: again...survivors are life-long carriers, usually
rainbow trout, brown in Europe
Transmission: horizontal through water, virus can occur
on eggs spawned by carriers, IP injection, birds, hatchery
equip

Viral Hemorrhagic Septicemia (VHS)

Pathogenesis: infection results in viremia, disrupts many
organ systems, 200-300g fish most affected
Environmental factors: low temp (< 8oC, 46oF)
External pathology: lethargy, hanging downward in water
(dropsy), swimming in circles, exopthalmia, dark
discoloration, hemorrhages in roof of mouth, pale gills
w/focal hemorrhages

Viral Hemorrhagic Septicemia (VHS)

Internal pathology: gut devoid of food, liver pale,
hemorrhages in connective tissue, kidney gray and swollen
(chronic), red and thin (acute)
Histopathology: necrosis of liver, kidney nephrons,
spleen, pancreas, melanin in kidneys and spleen (OUCH!)
Isolation/tests: isolated from kidney/spleen,
epizootiological evidence, definitive test is serum
neutralization, or FAT.

Viral Hemorrhagic Septicemia (VHS)

External hemorrhages

Liver red in acute stage

Viral haemorrhagic septicaemia in rainbow trout.

Note pale color of stomach region, pinpoint
haemorrhages in fatty tissue, and pale gills

Source: T Hstein

Viral haemorrhagic septicaemia in rainbow trout.

Note swollen stomach and pop eye

Type

Prevalent host type and location

I-a

Farmed rainbow trout and a few other freshwater fish in continental

Europe[10]

I-b

Marine fish of the Baltic Sea, Skagerrak, Kattegat, North Sea,

Japan[1]

I-c

Farmed rainbow trout Denmark

I-d

Farmed rainbow trout in Norway, Finland, Gulf of Bothnia

I-e

Rainbow trout in Georgia, farmed and wild turbot in the Black Sea[11]

II

Marine fish of the Baltic Sea

III

Marine fish of the British Isles and northern France, farmed turbot in
the UK and Ireland, and Greenland halibut (Reinhardtius
hippoglossoides) in Greenland[12]

IV-a

Marine fish of the Northwest Pacific (North America), North

American north Atlantic coast,[13] Japan, and Korea[1][14]

IV-b

Freshwater fish in North American Great Lakes region[14]

(3 ) Infectious Hematopoietic Necrosis

(IHN)
Who: sockeye, chinook, rainbows; cohos resistant
When?: 1950s in Oregon hatcheries. 100 million
mortalities between 1970-1980, if infected, 70%
mortality likely, esp. in young fish (fry: 90-95%
mort. possible)
What?: bullet shaped rhabdovirus, non- segmented
SS-RNA, sensitive to heat and pH, glycoprotein is
spiked on surface of virus

Viral Hemorrhagic Septicemia

Prevention: clean broodstock, water = fish, avoid infected
broodstock, test and slaughter

Can spread very quickly from farm to farm: avoid close

proximity to other farms
Vaccines are under development.
One EPA-approved disinfectant: Virkon AQUATIC
(made by Dupont). Bleach kills the VHS virus.

Infectious Hematopoietic Necrosis (IHN)

Reservoirs: survivors life-long carriers, adults shed virus
at spawning
Transmission: horizontal, primary mode is vertical via
ovarian fluid (virus hitches ride on sperm into egg);
however, feces, urine, and external mucus possible. Also,
feeding and inoculation have worked experimentally
Pathogenesis: gills suspected; incubation period depends
on temp, route, dose, age; extensive hemorrhaging,
necrosis of many tissues; death usually due to kidney
failure

Infectious Hematopoietic Necrosis (IHN)

Environmental factors: temp. very important,
slows below 10C, holding in tanks/handling
increase severity
External pathology: lethargy, whirling, dropsy,
exopthalmia, anemia, hemorrhaging of
musculature/fins, scoliosis
Internal pathology: liver, kidney, spleen pale;
stomach/intestines filled with milky fluid;
petechial hemorrhaging
Histopathology: extensive necrosis of
hematopoetic tissue of kidney/spleen

Infectious Hematopoietic Necrosis (IHN)

Definitive diagnosis: serum neutralization, FAT, ELISA
Prevention: avoidance, quarantine, clean water with UV,
ozone, virus-free stock; test, slaughter, disinfect; disinfect
eggs; vaccines under development; elevated water temp
No vaccines as of June 2007.

(4) Channel Catfish Virus Disease (CCVD)

Contagious herpes virus affecting only channel catfish less
than four months old
Occurs in SE United States, California, Honduras
Acute hemorrhagia, high mortality, first discovered in
1968
Agent: enveloped capsid, icosohedral nucleocapsid with
162 capsomeres
Physio/chemical properties: easy to kill, sensitive to
freeze-thaw, acid, ether, etc.

Channel Catfish Virus Disease (CCVD)

Environmental factors: optimal temperature 28-30C,
common during warmer months, cooler water = big
difference
epizootiology: horizontal, vertical suspected
external pathology: spiral swimming; float with head at
surface; hemorrhagic fins, abdomen; ascites; pale or
hemorrhagic gills; exophthalmia
Ipact 95 % ortality ( On Larva and fingerling)
Adlt as a carrier

Channel Catfish Virus Disease (CCVD)

Internal pathology: hemorrhages of liver, kidney,
spleen, gut, musculature; congestion of
mesenteries and adipose
Histopathology: necrosis of kidney, other organs;
macrophages in sinusoids of liver, etc.;
degeneration of brain
Presumptive diagnosis: clinical signs,
epizootiological evidence

Channel Catfish Virus Disease (CCVD)

Prevention: avoid potential carriers (survivors) or infected
fry, keep temperature below 27oC (will still produce
carriers), anage water qality and sanitation, cleaning the stf
with chlorine attenuated vaccine shows some promise
Therapy: none available...

Channel Catfish Virs Disease

Ephitelioma Populasum (cacar virus)

Disebabkan oleh kualitas air yang buruk,
padat tebar yang tinggi

target : ikan jenis carp

Gejala : mncl bintik ptih susu berisi
nanah yang menyebar sangat cepat
mntp selurh permukaan tubuh dgn
ketebalan 1-2 mm.

Pengobatan
Larutan arsenik 1 % dalam
senyawa arycil. Penyuntikan
pada perut ikan. Lakukan 3 kali
penyuntukan selanjutnya
dengan 5 % arsin dalam
senyawa arycil

Lymphocystis
Disebabkanoleh iridovirus/ DNA
Dapat menyerang ikan air twar maupun laut
Gejala: Terdapat penebalan pada sel jaringan ikat
sehingga mncl tonjolan atau nodul pada ikan bisa
berkelopok ata sendiri. Sasaran uttama kulit atua
insang.
Tidak ada pengobatan yg efektiv, hanya
pencegahan melalui menjaga kualitas air dan
peningkatan imunitas ikan. Penularan melalui
makanan atapun suntikan dan air.

Koi Herpes Virus (KHV)

Disebakan virus DNA dari strain herpes viridae
menyerang ikan mas (Cyprinus)dan koi
muncul : temperatur turun dan kualitas air buruk
Gejala : nafsu makan turun, ikan megap
megap,terdapat keruskan insang ( bintik putih dan
merah pada insang)
menyerang insang dan ginjal ikan sehingga
terbentuk inflamasi pada ginjal, kllit
melepuh,insang membsk, kematian 1-5 hari.
Sering diikuti infeksi sekunder oleh bakteri dan
jamur ketika luka ada di permukaan tubuh

Pengobatan
Segera dipindahkan ikan yang sakit (isolasi) ke
suhu air yang lbh tinggi ( 30 C) selama beberapa
hari kemudian suhu diturunkan
sampai 27 C keumdian dikeluarkan. Dilanjtkan
pengobatan seknder. Jika sdah mewabah sebaiknya
dimusnahkan
Pencegahan : padat tebar dikurangi, penggunaan
benih yang berkalitas dan bebas KHV, mnjaga
kualitas air, pemberian pakan yg baik dan sehat,
desinfeksi pada semua perslatan dan kolam,
mnitoring dan evaluasi ikan dan lingkngan

Viral Nervous Necrosis (VNN)

Penyebab : Betanodaviruses family Nodoviridae
Menyerang larva ikan sampai dewasa (ikan laut)
Target serangan : sistem syaraf dan retina
Gejala : Nafsu makan turun, berenang yang tidak teratur,
warna tubuh menjadi gelap, posisi terapung dengan perut
diatas karena pembengkakan, Kerusakan pada retina
mata.
Menyebkan kematian 50 %
Penyebaran dengan horisontal maupun vertikal
Vaksinasi diperlukan untuk mencegah VNN
Pencegahan : manage Kualitas air, benih, pakan dan
lingkungan serta sarana budidaya.

However, you can always take precautions!