Koi herpesvirus

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					                                                       Introduction

                                                 • Koi herpesvirus is a recently
      KOI HERPESVIRUS: USING                       emerged viral disease of carp (Cyprinus
                                                   carpio) in all of its varieties
         BIOSECURITY AND
                                                 • First officially identified in 1998.
         SURVEILLANCE TO
                                                 • Examination of archive material indicated
      PRODUCE DISEASE FREE                         KHV presence as early as 1996
               FISH                              Poorly regulated international trade of
                 David Huchzermeyer
             Sterkspruit Veterinary Clinic         ornamental carp (Koi) and goldfish
                     Lydenburg




                                                     Major impact on commercial
       Worldwide distribution
                                                        food carp production
  molecular comparison of diverse                •    Japan
  geographic isolates indicated that these       •   South East Asia
  represented a largely homogenous               •   North America
  group with only slight differences
                                                 •   United Kingdom
  between Western and Asian isolates
                                                 •   Poland
                                                 •   KHV recently reported in confiscated illegal
                                                     koi imports in the Philippines




                                                                     Spring viraemia of
                   History
                                                                        carp (SVC)
• In the past international movement of carp
  was governed by concerns over the spread       • Rhabdovirus
  of spring viraemia of carp (SVC)               • Can survive outside of the host in fresh
• Health certification requirements applied to     water for up to 5 weeks at 10oC
  carp but not to koi.                           • Disease of cyprinids (carp, goldfish and koi)
• Spread of SVC in recent years has also been    • Spread from the European continent
  blamed on movement of koi                      • 1988 – Brazil             2002 – USA
                                                 • 2004 – China              2006 - Canada




                                                                                                    1
   Introduction of exotic diseases threatens:-
• natural aquatic species diversity                            Family Herpesviridae.
• livelyhood of subsistence and commercial              Cyprinid herpes virus 1 - carp pox herpes virus
  fishermen
                                                        Cyprinid herpes virus 2 – herpes viral
• aquaculture development and investment                  haematopoietic necrosis virus of goldfish
                                                        Cyprinid herpes virus 3 - Koi herpes virus
• employment opportunities
                                                        CyHV-1, CyHV-2 and CyHV-3 have been shown to
• local economies                                         be closely related to each other and distantly
                                                          related to channel catfish virus (IcHV-1)




                                                               KHV and other species
       Carp pox lesions CyHV-1
                                                        • Recent evidence indicates that gold fish
                                                          (Carassius auratus) may act as
                                                          asymptomatic carriers of the koi herpes
                                                          virus
                                                        • No information available on susceptibility
                                                          of African cyprinids




                                                           History of a typical outbreak
         Manifestation of disease                       • Clinical signs are not specific.
  • Temperature dependant- permissive temperature       • Incubation period 7 days or less
    for infection lies between 16 and 25oC              • Rapid onset of mortality. Within 24 to 48
  • Disease follows introduction of stressed infected     hours of onset of clinical signs. (82 % or
    fish into naïve populations or after contact with
    strange fish                                          more of exposed fish at 220C can die within
  • Virus enters host through the skin. Frequent          15 days)
    netting and high stocking density contribute to     • 80 to 100 % mortality
    rapid spread
  • susceptibility and mortality are greatest between
                                                        • All ages affected, but fingerlings more
    22 and 25oC                                           susceptible than older fish




                                                                                                           2
                                                      External signs of disease
                   Symptoms
                                                  • External signs in infected fish are non-specific but
                                                    include
   •   Affected fish remain near the surface      • sunken eyes,
   •   Swim lethargically                         • hyperaemia and discolouration of the skin,
   •   Respiratory distress                       • fin erosion,
                                                  • thickening and loss of surface mucus.
   •   Aberrant swimming behaviour
                                                  • gills may take on a characteristic mottled
   •   Sick fish may still be attracted to feed     appearance, with visible white or brown streaks of
                                                    dead gill tissue.
                                                  • Changes in the internal organs are inconsistent and
                                                    non-specific




            Gill necrosis (KHV)                      An increase in opportunistic
                                                   parasites in KHV infected fish can
                                                        be diagnostically misleading
                                                  • Ichthyobodo necatrix
                                                  • monogenean flukes
                                                  • bacterial infections with Flavobacterium
                                                    columnaris




        KHV - need for certified
                                                                     Diagnosis
          pathogen free fish
• Difficulty of identifying carrier fish          • Opportunistic infections may mislead a
• Unquantifiable risk associated with:-             diagnosis of KHV
     naturally resistant fish
     survivor fish                                • PCR testing of gill and kidney swabs will
     „vaccinated‟ fish                              reliably confirm active infection
• Access to export
  markets




                                                                                                           3
           Direct diagnostic tests                                    Current diagnostic limitations
                                                                   • As with other herpes viruses, an asymptomatic
1. PCR (polymerase chain reaction) detection                         carrier state appears to exist and recrudescence
  of viral DNA.                                                      may occur. PCR techniques in use in South Africa
                                                                     are unable to identify carrier fish and a negative
2. histopathology                                                    PCR result from a healthy fish does not rule out
3. virus isolation on koi tissue culture. The                        KHV infection.
  virus is difficult to isolate on conventional                    • Reliable tests for virus isolation and tests to
  fish tissue cell cultures and culture based                        identify antibodies to the disease have been
                                                                     developed elsewhere but are not yet available in
  tests are not available in South Africa                            South Africa.




        Diagnostic confirmation                                            Effect of temperature
• Indirect diagnostic test                                         • Temperature is the predetermining factor
    ELISA to detect antibody to KHV.                                 that controls whether KHV develops into
   Limitations:                                                      lethal infection.
 - although a positive test indicates previous                     • Peak mortality at permissive temperature of
   exposure, it gives no indication of whether                       22 – 250C.
   the fish is still infected.                                     • KHV outbreak can result in up to 98 %
 - unknown how long antibodies persist                               mortality




                   Other factors                                           Diagnostic limitations
•   host susceptibility              Gill epithelial hyperplasia   • Where KHV is causing mortality the current
                                    typical of chronic ammonia
•   transport stress                          poisoning              diagnostic techniques are highly accurate in
•   stocking density                                                 identifying the virus.
•   water quality – ammonia                                        • Once fish have recovered or if they have
                                                                     been exposed but are held outside of the
                     gas super-saturation                            permissive temperature it may be difficult
                                                                     to detect the presence of the virus.




                                                                                                                          4
   How can we limit the risk of                      Proposed strategies to limit
       spreading KHV?                                       KHV losses
                                                 • avoidance of exposure particularly at
• No single or group of measures will give         shows,
  absolute certainty that a single fish or a     • development of vaccines,
  group of fish are free from Koi herpesvirus    • use of thermal temperature regimes
  unless the fish originate from a certifiable   • selective breeding for increased
  KHV free population                              resistance and
                                                 • use of resistant hybrids in fish
                                                   production




          KHV Regulations                          Virus free hatchery protocol
                                                    was developed based on:
                                                 • the 95% confidence level for detecting a disease
• KHV has been included in the OIE Aquatic         with a 2% prevalence in a population of fish
  Animal Health Code as a notifiable disease       exceeding 1000 individuals with 6 monthly testing
                                                   over a 2 year period and
• EU import certification requirements have      • on the assumption that vertical transmission would
                                                   not occur
  been amended to include guarantees for
  freedom from KHV by some EU member             • no other cyprinid species would be allowed on the
                                                   farm
  countries




  Breeding certified KHV free                     Prerequisites for a disease free
      fish for the market                                     status
                                                 • Closed population of fish
                                                 • Closed water supply
                                                 • Any newly introduced fish must originate from a
                                                   source with the identical or higher standard of
                                                   disease free certification
                                                 • Disease free status based on EU directives and on
                                                   principles laid down by the International Aquatic
                                                   Animal Health Code (OIE)




                                                                                                        5
     Prevent transmission from                 Hormonal induction - GnRH
         brood fish to ova                            analogue
•   Only healthy brood fish were selected
•   Hormonally induced artificial spawning
•   Artificial fertilization of ova
•   Disinfection of fertilized ova
•   Incubation of ova in isolation hatchery
•   Raising fry and fingerlings in isolation
    ponds




              Collecting milt                      Stripping ripe eggs




                  Fertilization                 Fertilized eggs placed on
                                                brushes or in Zoug jars




                                                                            6
Fertilized eggs water hardening                      Eggs moved to disinfectant
      prior to disinfection




                                                     Disinfected eggs being moved
      Egg disinfection - iodophore                   out of the brood fish facility
 disinfectant at 50 ppm for 10 minutes




                Grow out                           Principles of good biosecurity
• Fry were kept in hatchery tanks and initially   • Access control
  fed Artemia spp. nauplii before being           • Brood fish separated from hatchery and grow out
  released into recently filled and fertilized      facility
  earthen grow-out ponds.                         • Separate staff designated to each facility
• The grow-out ponds were used on an “all-in      • Closed water supply
  all-out” basis with each pond being             • Fenced and net covered ponds
  emptied, disinfected and dried between use.     • Strategic disinfection
                                                  • All in all out stocking




                                                                                                      7
  6 monthly disease free testing
           procedure                                  Sample submission from a minimum of
1. After harvesting of earth ponds, fish were            150 fish destroyed humanely for
  placed into holding ponds for observation for                     sampling
  at least 2 weeks
2. 400 - 800 fish were packed into bags with
  oxygen and subjected to normal transport           • Spleen, liver and kidney in phosphate
  stress                                               buffered saline on ice for Rhabdovirus
3. Further quarantine in recirculated facility for     carpio virus isolation on tissue culture
  minimum of 2 weeks at or near permissible
  temperature                                        • Gill swabs for PCR detection of Koi
                                                       herpesviral DNA
4. Ammonia levels allowed to build up during
  quarantine




       Replacement of original
                                                           Disease free certification
             broodstock
                                                     • Following the initial two-year period with
   Over the following 2 years the original             six monthly tests the fishery was able to
   broodstock was replaced with offspring              meet EU import requirements for cyprinids,
   from the tested population                          providing the necessary guarantees for
                                                       freedom from Rhabdovirus carpio as well
                                                       as from KHV.




                 Thank you




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posted:10/19/2011
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