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Disease challenge models




Dr Grant Stentiford

European Community Reference Laboratory for Crustacean Diseases

Pathology and Parasite Systematics team, Cefas
Diagnosing disease in crustaceans



 Research/diagnostics tools are similar for use with other animal groups
 Histology provides a front-line tool for observations of deviations from normal
 Electron microscopy provides information on pathology/pathogens/life cycles
 Antibodies are available against most major crustacean pathogens
 Molecular diagnostics (e.g. PCR) are widely used to confirm pathogen type
 No reliable cell lines are available (e.g. for viral culture)
 Generally rely on a combination of the above for successful diagnosis
Diagnostic accreditation and in-house expertise




 Histology: ISO 17025 Method for testing crustaceans for
  pathogens and notifiable diseases by histological analysis
 Molecular diagnostics: ISO17025 Flexible scope
  accreditation for RT-PCR and PCR of fish, shellfish and
  crustacean pathogens
 Experimental Facility: GLP accredited facility. Biosecure
  containment facility for exotic pathogen work


 Capacity to work with a wide range of host species and all
  globally significant pathogens (e.g. EC Directive listed,
  OIE listed, emerging pathogens)
Typical challenge studies


Q. Are crustaceans from EU waters susceptible to WSSV?
Susceptibility testing
Viral challenge studies


Q. Are crustaceans from EU waters susceptible to WSSV?


                                                                                  Cumulative mortality of P. vannamei injected with edible crab innoculum




                                                                              5


                                                                              4




                                                             Mortality rate
                                                                              3


                                                                              2
                                                                                                                                                  control mortalities

                                                                              1                                                                   Injected mortalities
                                                                                                                                                  Fed mortalities

                                                                              0
                                                                                   1           2             3            4           5             6                   7
                                                                                                                        Day




Injection, feeding
or bath exposure
   to innoculum
  prepared from             Injection of filtered       Mortality,
  frozen WSSV-            homogenate from WSSV       histopathology,
 infected shrimp            exposure trial using    TEM and WSSV
                             European species        PCR of shrimp
                                                        recorded
Viral challenge studies  e.g. European lobster




                                             WSSV-Infected cuticular epithelium of H.gammarus



  Experimental infection of European lobster (Homarus gammarus) via feeding
   of WSSV-infected P. vannamei
Current work areas




     Diseases in fisheries    Diseases in aquaculture   Transboundary movements




Immuno-competence toolbox    Crustaceans as sentinels   Pathogen systematics
Visit www.crustaceancrl.eu for information on our diagnostic remit and research programme

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Disease Challenge Models - APEG

  • 1. Disease challenge models Dr Grant Stentiford European Community Reference Laboratory for Crustacean Diseases Pathology and Parasite Systematics team, Cefas
  • 2. Diagnosing disease in crustaceans Research/diagnostics tools are similar for use with other animal groups Histology provides a front-line tool for observations of deviations from normal Electron microscopy provides information on pathology/pathogens/life cycles Antibodies are available against most major crustacean pathogens Molecular diagnostics (e.g. PCR) are widely used to confirm pathogen type No reliable cell lines are available (e.g. for viral culture) Generally rely on a combination of the above for successful diagnosis
  • 3. Diagnostic accreditation and in-house expertise Histology: ISO 17025 Method for testing crustaceans for pathogens and notifiable diseases by histological analysis Molecular diagnostics: ISO17025 Flexible scope accreditation for RT-PCR and PCR of fish, shellfish and crustacean pathogens Experimental Facility: GLP accredited facility. Biosecure containment facility for exotic pathogen work Capacity to work with a wide range of host species and all globally significant pathogens (e.g. EC Directive listed, OIE listed, emerging pathogens)
  • 4. Typical challenge studies Q. Are crustaceans from EU waters susceptible to WSSV?
  • 6. Viral challenge studies Q. Are crustaceans from EU waters susceptible to WSSV? Cumulative mortality of P. vannamei injected with edible crab innoculum 5 4 Mortality rate 3 2 control mortalities 1 Injected mortalities Fed mortalities 0 1 2 3 4 5 6 7 Day Injection, feeding or bath exposure to innoculum prepared from Injection of filtered Mortality, frozen WSSV- homogenate from WSSV histopathology, infected shrimp exposure trial using TEM and WSSV European species PCR of shrimp recorded
  • 7. Viral challenge studies e.g. European lobster WSSV-Infected cuticular epithelium of H.gammarus Experimental infection of European lobster (Homarus gammarus) via feeding of WSSV-infected P. vannamei
  • 8. Current work areas Diseases in fisheries Diseases in aquaculture Transboundary movements Immuno-competence toolbox Crustaceans as sentinels Pathogen systematics
  • 9. Visit www.crustaceancrl.eu for information on our diagnostic remit and research programme