Aquatic Animal Diseases

(global trends & spread & emerging threats)

• The spread of diseases is the most feared threat to aquaculture. It is a
matter of global concern especially with increased trade & movement of
live aquatic animals & their products across national borders. Examples
include:
• White spot syndrome disease in shrimp spread to 22 countries via
trade in post-larvae
• Taura syndrome spread from Americas to Asia via live shrimp
movements
• Gyrodactylus salaris spread from Sweden to Norway via live
juvenile salmon for stock enhancement
• First case of Sleeping disease in UK was linked to imported trout
fillets
• EHN virus spread from Germany to Finland via live farmed
sheatfish imports
• First cases of SVC in Switzerland, USA, Denmark were linked to koi
carp imports
• Koi herpesvirus has been linked to international koi carp trade
• ISA outbreaks in Atlantic salmon in Chile in 2007: ISA virus was
most similar to isolates from Norway.
Aquatic Animal Diseases (global trends & spread & emerging threats)
 Aquatic Animal Diseases
(global trends & spread & emerging threats)
• Viral haemorrhagic septicaemia (VHS)
• Pancreas disease (PD)
• Cardiomyopathy syndrome (CMS)
• Heart and skeletal muscle inflammation (HSMI)
• Infectious salmon anaemia (ISA)
Viral diseases of shrimps
Viral diseases of shrimps
Currently, at least 14 virus diseases of cultured shrimp are
recognised.
The major virus families present in the crustaceans include
Parvoviridae, Baculoviridae, Picornaviridae, Reoviridae, Togaviridae,
Cornaviridae.
The parvo-like viruses
The parvo-like viruses are characterised by their small size
(20 – 30 mm), single stranded DNA genome and unique
cytopathology.

1a) Hepatopancreatic parvo–like virus (HPV)

• HPV has been widely distributed in indo-pacific.
• In cases of dual infections, mortality range from 50-100%
• HPV infections - characterized by atrophied and white
hepatopancreas, reduced growth rate, anorexia, poor
preening activity, and opacity of abdominal muscle,
surface and gill fouling and secondary infections by
opportunistic pathogens like vibrio spp.
Hepatopancreatic parvo–like virus (HPV)
• Histologically -single, prominent, basophilic, Feulgen-
positive intranuclear inclusion bodies in hypertrophied
nuclei of hepatopancreatic tubule epithelial cells.
• Lateral displacement of nucleolus and margination of
chromatin
• In the early stages, HPV inclusions are eosinophilic bodies
centrally located in the nucleus associated with nucleolus.
• In Indian shrimp farms, the HPV shows a low incidence
rate.
HPV
Infectious hypodermal and hematopoietic necrosis virus
(IHHNV)

• smallest DNA virus of the penaeid shrimp - diameter 22 nm.

• IHHNV is distributed worldwide with more prevalence in the Southeast Asia. P.
monodon has been found to be the natural host of the virus.
• IHHNV causes catastrophic epizootics in cultured juveniles of P. stylirostris.
• Affected shrimp exhibit reduced growth, cuticular deformities to rostrum and other areas
of exoskeleton.
• The infection is transmitted both horizontally and vertically. In P. monodon, the infection
causes bluish coloration and opaque abdominal musculature.
IHHNV
578
Infectious hypodermal and hematopoietic necrosis virus
(IHHNV) - 2

• Histologically, confirmative diagnosis - prominent Cowdry type A eosinophilic

intranuclear inclusions
• IHHN results in hypertrophic nuclei of cells of ectodermal (epidermis,
hypodermal epithelium of fore and hind gut, nerve cord and nerve ganglia) and
mesodermal origin (hematopoietic organs, antennal gland, gonads, lymphoid
organ, connective tissue and striated muscles).
• Survivors of IHHN epizootics carry the virus for life and transmit the virus
horizontally and vertically.
Lymphoid organ parvo-like virus (LOPV)
LOPV was found in cultured shrimps (P. monodon, P. merguiensis
and P. esculentus) in Australia.
Affected shrimps have multinucleated giant cells in their
hypertrophied lymphoid organs. Giant cells showed nuclear
hypertrophy, marginated chromatin and formed fibrocyte-
encapsulated spherical structures.
Giant cell nuclei had intranuclear inclusion bodies containing
DNA. Electron microscopic studies revealed the presence of 25-
30 mm diameter viruslike particles. There is a speculation that
IHHNV and LOPV are the same agent.
Reolike viruses

Reo-like viruses are reported from P. japonicus, P. monodon, P.

chinensis and P. vannamei.
Hepatopancreas has been suggested as the principal target for
both the viral strains.
Presumptive diagnosis of infections can be made with routine
histology and staining. Feulgennegative cytoplasmic inclusion
bodies are demonstrated in I & R type cells of atrophied
hepatopancreas. Transmission electron microscopic (TEM)
pictures show crystalline array of virus particles of 50-60 mm
diameter.
Reovirus infections were always reported in mixed infections.
Hence the role of reovirus as pathogens is not completely clear.
REO-VIRUS 580
Inclusion body of reovirus
Togaviruses
Infects lymphoid organs of many shrimps

Histological lesions. Extreme hyperplasia and

metastasis of the lymphoid organ (Oka organ) The
virus particles (30 nm) was observed in
intracytoplasmic eosinophilic to pale basophilic
cytoplasmic inclusion bodies occurring in cells with
highly vacuolated cytoplasm.
• Cells were found to have pyknotic and karyorrhetic
nuclei.
Baculoviruses
• (1) Type A occlusion body forming viruses BP and
MBV and
• (2) Type C nonoccluded baculoviruses BMN, TCBV,
Owen’s hemocyte–infecting baculovirus and WSDV.
• Transmission: horizontal, some transmit vertically
• In hatcheries, BP and BMN often cause serious
epizootics in larvae and postlarvae (PL)
• MBV produce serious infections and mortalities in the
late PL and juvenile stages of hosts.
BP Type baculoviruses
• BP (Baculovirus penaei): diagnosis: demonstration of
prominent tetrahedral occlusion bodies in unstained
squash preparations of hepatopancreas, midgut or
faeces and also in histological sections.
• In histological sections, occlusion bodies are found in
single or multiple, eosinophilic, usually triangular
within hypertrophied nuclei of hepatopancreatic
tubule epithelial cells or in midgut epithelial cells.
• BP has not been reported outside of the Americas and
Hawaii
BP
573
Monodon type baculoviruses
• MBV enjoy a world-wide distribution. These are type
A baculoviruses measuring 75 x 324 mm.
• Diagnosis: presence of single or multiple, generally
spherical intranuclear occlusion bodies in
hepatopancreas and midgut epithelial cells. Squash
preparation (0.05% aqueous malachite green),
epifluorescence microscopy and acridine orange
staining visualizes MBV occlusions
MBV
574
• MBV was first discovered in a quarantined population
of P. monodon that had originated from Taiwan.
• Despite the world distribution, MBV is not a highly
virulent pathogen of P. monodon. MBV is found in
healthy prawns and in disease epizootics, P. monodon
has been found to frequently have mixed infections by
MBV and other viral, bacterial or protozoan
pathogens.
BMN (Baculoviral midgut gland necrosis virus) and
other type C baculoviruses
• Does not produce occlusion body in the nuclei of
infected cells.
• Diagnosis: necrotic hepatopancreatic (midgut gland)
tubule, epithelial cells that have hypertrophied nuclei
with marginated chromatin, diminished nuclear
chromatin, nucleolar dissociation and no occlusion
bodies.
• Type C baculoviruses are enveloped viruses of 72-310
nm and may occur in dense aggregates especially near
to nuclear membrane and surrounding virogenic
areas.
BMN
576
BMN (Baculoviral midgut gland necrosis virus) and
other type C baculoviruses..2
• BMN causes serious epizootics in hatchery reared P.
japonicus larvae in Southern Japan. Causes sudden
onset and high mortality rate. The disease subsides by
PL 20.
• Diagnosis: The infected larvae float inactively on the
surface and have a white turbid midgut line through
the abdomen.
• Histological conformation: presence of necrotic
hepatopancreatic tubule epithelial cells with
hypertrophied nuclei having marginated and
diminished chromatin, nucleolar dissociation and the
absence occlusion bodies.
White spot syndrome virus (WSSV)
• WSSV, formerly known as SEMBV is a nonoccluded
baculovirus – like agent
• Epizootic of white spot disease cause mortalities ranging
up to 80  100% in 2  7 days
susceptible species: Ongrowing juvenile shrimp of many
species of all ages but mostly from 1 - 3 months old in the
grow-out ponds.
• WSSV outbreak occurs in all types of farming systems
irrespective of stocking density, water quality and salinity.
Diagnosis: Infected shrimp swim to the surface and gather at
the pond dykes.
• Broken antennae, white spots of 1 mm size in the cuticle
and / or reddish discoloration, empty guts and cuticular
epibiont fouling and lymphoid organ swelling.
White spot syndrome virus (WSSV)..2
• Histological signs:
• widespread and severe nuclear hypertrophy,
• chromatin margination,
• eosinophilic to large basophilic intranuclear
inclusions
• variable multifocal necrosis in most tissues of the
animal.

• The virus is enveloped, rod shaped to elliptical and

measured 292 x 111 mm. The white spot disease could
be controlled by avoidance of infection and
contamination.
WHITE SPOT DISEASE
Yellow Head Virus (YHV)
• YHV is an RNA virus reported only from P. monodon
in Thailand.
• All ages of juveniles could be infected and mass
mortalities up to 100% are observed within 3 - 5 days
• Diagnosis: Pale body colour with yellowish gills and
hepatopancreas. It affects many tissues such as gills,
lymphoid organ, hemocytes and connective tissue.
• Histology: Degenerative changes in nuclei and
presence of cytoplasmic basophilic inclusion bodies.
Management of shrimp disease
1. Early and effective detection of pathogens by
improved diagnostic methodologies
2. Use of biosecure, closed or semiclosed recycle
system with reduced water exchange
3. Maintain good water quality and treat water before
use especially for hatcheries.
4. Construct reservoirs for storing water without
directly taking from the sea and treat reservoir water
5. Bio-security approach for preventing pathogen
introduction by screening entry of infected hosts,
carriers like crabs, fish or birds, stray dogs, avoiding
contaminated inanimate objects
6. In case of a disease outbreak, disinfect contaminated
water before discharge.
7. Maintain good pond preparation by sun drying pond
bottom, desilting and ploughing. Application of 100
ppm CaO (burnt lime).
8. Avoid over stocking to reduce the stress on the
animals and chances of water quality deterioration
9. Production of specific pathogen free (SPF) shrimps and
the development of specific pathogen resistant (SPR)
strains
• SPF animals are produced by selecting animals free of
known and detectable pathogens and raising them under
controlled and strict sanitary conditions.
• The SPR animals are developed through selective breeding
of animals known to be less susceptible to specific
pathogens.
• This is useful for production of high health (HH) post-
larvae, free of, or resistant to, known pathogens.
• Reports indicate that Penaeus merguiensis has a higher
resistance to WSSV than P. monodon.
• However, in certain cases SPF stocks may perform poorly
when exposed to pathogens in the field.
10. Feed nutritionally balanced diet avoiding excess feed.
11. Use of immunostimulants and vaccines
12. Development of co-operative disease control strategies.
Co-operative program involving farmers, health care
specialists, scientists and government agencies to track the
movement of a potential pathogen, for notification of
neighbouring farms of an infection and proper disinfection
of the infected stock and water before discharge
13. Locate hatchery away from the shrimp farm to prevent
cross contamination of hatchery source-water and air borne
contamination of pathogens from the infected farm.
14. Use only virus-free broodstock to prevent vertical
transmission of the viral diseases in particular.
15. Avoid importing of broodstock and larvae.

16. Maintain broodstocks of different sources in

separate holding tanks and rear the larval population
in separate batches to prevent cross contamination.
17. Do not use trash fish and shellfish to feed
broodstock which can act as carriers of viral
pathogens. In case of feeding with trash fish, care has
to be taken for properly cooking the feed before use.
What is KHV?
• Cyprinid herpesvirus-3 or CyHV-3
• DNA virus – family Herpesviridae
• Susceptible species – Common carp (Cyprinus carpio carpio), Koi
(Cyprinus carpio koi) and Ghost carp (C. carpio goi)
• Resistant species – common goldfish (Carassius auratus) and grass
carp (Ctenopharyngodon idella)
KHV mortality patterns:

• 80 – 90% mortality in susceptible populations

• Clinical disease occurs must commonly when water temperatures are
between 22 – 27C (72 – 81F)
• However new cases have been reported in lower water temperatures
down to 18C (64F)
Signs of KHV:
• Non-specific
• Gill mottling with white necrotic patches dispersed between normal
red gill tissue
• Bleeding gills
• Sunken eyes
• Skin lesions
• Notched nose
Methods of transmission:
• Direct contact with infected fish
• Direct contact with fluids from infected fish
• Direct contact with water or mud which infected fish had been
exposed to
• Fomites/vectors from contaminated systems
How do fish become infected?
• Infective virus enters susceptible fish through gills, skin and possibly
gut
• Depending upon water temperature, susceptible fish that get exposed
may become infected and die
• OR may survive the initial outbreak and become carriers
Latent carriers
• Survivors of initial infection typically never show clinical signs again
• However they are still infected and can spread disease to susceptible
fish
• This latent carrier possibility is what makes KHV especially
problematic to commercial dealers *
Water temperature and KHV:
• Clinical KHV disease has an average incubation period of 14 days
following naïve fish exposed to infected or carrier fish
• Most mortalities occur between 22 and 25.5C (72 -78F)
• Virtually no mortalities above 30C (86F)
• New cases have shown clinical outbreaks in temperatures below 21C
(70F)
How to confirm KHV infection:
• No one can tell by just looking at the fish!
• Laboratory tests are required
• Direct methods – actually detect the virus or “pieces” of its genetic
material
• Indirect methods detect antibodies produced by the fish after
exposure to the virus
Direct methods
• Virus isolation (growing the virus) on KoiFin (KF) cell lines
• Polymerase chain reaction (PCR)
• Best samples from alive then euthanized fish – posterior kidney
• Non-lethal samples of blood, feces or gill clips can be used but results
are often less reliable
Indirect methods:
• Enzyme-linked immunosorbent assay (ELISA)
• Virus neutralization (VN)
• Non-lethal sample using blood
• A positive ELISA or VN indicates that a fish has antibodies ( protective
immune product) against KHV virus*
No test answers all KHV questions!
• Negative tests by either direct or indirect methods do NOT mean fish
are not carriers
• No test definitely detects all carriers or survivors of outbreaks
• Using diagnostic tests as a part of a combined preventive strategy is
currently the most practical and cost effective
Why can’t the diagnostic tests give a definitive
answer?
• PCR is the preferred test for confirmation of active clinical infection –
fish are sick and dying
• PCR can detect some latent carriers but not all – surveillance
screening of “normal” populations OR fish in quarantine
• PCR is the most economical and easiest to implement
What about the antibody tests?
• Fish that have antibodies against KHV mean that they have been
exposed to the virus at some time in the past
• Two possibilities – active infection with outbreak just starting OR
latent carrier
• Antibody producing immune cells take time to ramp up with new
infections – false negatives
• Same cells may slow down or stop antibody production if non-lethal
infection occurred a long time ago and fish no longer sick – false
negative on carrier state
Individual vs population screening:
• Antibody testing is the preferred method for detecting carrier status
in valuable individuals *
• But what about testing groups of 10, 100 or 1000 fish?
• Unless you test them all by antibody detection methods and get all
true negative results each time – can NOT be 100% sure
How to prevent KHV:
• Quarantine (Q)
• Water temperature in between 21 – 27C (70 – 80F) …… 24C (75F)*
• 30 day minimum time period
• “Reverse quarantine” after 30 days initial Q period
• Diagnostic screening tests
Quarantine
• Definition
• Separate tank or pond w/ separate
water & filtration system
• Preferably located away from the
main pond
• Time Period
4-6 weeks
Can KHV be treated?
• No treatment for KHV!
• Virus can remain viable for 3 days in water without fish hosts
• Common disinfection protocols can eliminate the virus from
contaminated systems and equipment
Regulatory Concerns with KHV:
• 2007 – World Organization for Animal Health (OIE) added KHV to the
notifiable disease list for finfish
• USDA-APHIS asks accredited veterinarians and diagnostic labs to
report positive cases to the AVIC of the state in which the case
occurred
Regulatory Concerns with KHV:
• USDA-APHIS has no mandatory requirements on KHV status for koi
moving interstate or international
• No mandatory depopulation for positive KHV fish
• KHV now a Hawaii State reportable disease – positive cases must be
reported to the Hawaii State Veterinarian
Spring Viremia of Carp (SVC)
• RNA virus
• Affects koi & goldfish
• Mortality up to 70% in affected populations
• Temperature range
12-24°C
54-72°F
• OIE reportable disease
• FAD and mandatory depopulation required by both State and USDA-
APHIS
New USDA Guidelines for SVC
• 8 spp. of fish that are susceptible to SVC are now under USDA
oversight for importation into the U.S.
• Required documents
• USDA import permit (form VS-135)
• Veterinary health certificate from the exporting country
• USDA VS Port Veterinarian must inspect each shipment at entry port
Kelompok: Virus
Golongan: HPIK Gol I
Organisme Penyebab: Viral nervous necrosis (Beta
nodavirus)
Nama Penyakit: Viral nervous necrosis (VNN) atau Viral
encephalopathy and retinopathy (VER)
Sea bass fry suffering an outbreak of viral encephalopathy and
retinopathy or viral nervous necrosis (VER, or VNN) float and show
swimming ataxia mostly on the surface.
swim bladder over-inflation/distention is a common finding
in fish suffering VNN/VER caused by strains of piscine
Nodavirus; sea bass (left), shi drum (right).
Betanodavirus particles isolated from Atlantic cod, in
cytoplasm of susceptible (E-11) cells.