Viral reproductive disorders in sows<br />an update 2011<br />H. Nauwynck<br />Laboratory of Virology<br />Faculty of Vete...
1. CL (progesterone)<br />Normalgestation in the sow<br />2. Cytokine/growth factor <br />cross talk<br />?<br />3. Immune...
How do viruses cause reproductive problems?<br />General disease (fever) <br />	~ pro-inflammatory cytokines causing disto...
How do virusesreachembryos/fetuses? <br />1. Contaminatedsperm<br />
2. Viremia + transplacental spread<br />How do virusesreachembryos/fetuses? <br />
1. Via contaminatedsperm<br />Virus excretion in sperm<br />~ viremia<br />DNA-viruses<br />ADV (<14 dPI)<br />PPV (<14 dP...
At what stage can embryos be infected ?<br />	2-8 cells	morula	blastocyst	hatched blastocyst<br />(0-4dpins)	(4-5dpins)	(5...
Zona pellucida is an ideal barrier<br />1° Physical barrier<br />Channels in ZP : ± 30 nm (inner ø)<br />Viruses > 30nm (A...
Outcome of embryonic PPV-infection<br />Uninfected<br />PPV-infected<br />
Outcome of embryonic PCV2-infection <br />Surgical ET<br />HB 6 dpins<br />PCV2/mock<br />Inspection of <br />ovaries<br /...
Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />a...
Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />a...
Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />a...
2. Viremia + transplacental spread<br />DNA-viruses<br />(PPV, PCV2, ADV)<br />RNA-viruses<br />(PRRSV, CSFV, <br />PEV 1,...
Transplacental spread - cell free virus<br />PRRSV (> 70dpi)<br />PPV, PEV, EMCV<br />*<br />*<br />*<br />*<br />*<br />*...
Normal placenta<br />fetal site<br />maternal site<br />
Transplacentalspread- PRRSV - 1. cell free virus + free mø<br />
Transplacental spread - PRRSV - 2. cell free virus + migrating mø<br />
Transplacental spread - PRRSV - 3. infection of migrating mø<br />
Transplacental spread - PRRSV - 4. entry of fetal placenta by <br />migrating mø<br />
Transplacental spread - PRRSV - 5. explosive replication in fetal placental mø  <br />
Transplacental spread - PRRSV - 6. apoptosis in infected mø and bystander cells<br />
Transplacental spread - PRRSV - 7. apoptosis in infected mø and bystander cells/necrosis?<br />
Transplacental spread - PRRSV - 8. dysfunction placenta<br />
Transplacental<br />spread-<br />cell-associated virus<br />*<br />*<br />*<br />*<br />PCV2, ADV, CSFV <br />*<br />*<br ...
Cell-associated transplacental spread of ADV<br />Maternal<br />placenta<br />Fetal<br />placenta<br />Maternal<br />place...
Interfetal virus spread<br />PPV/PEV/PRRSV<br />ADV/CSFV<br />PCV2<br />*<br />
Clinical outcome - fetus <br />Infection time (days of gestation)<br />0-30<br />†<br />†<br />†<br />-<br />†<br />†<br /...
Clinical outcome - sow<br />Infection time (days of gestation)<br />60-90<br />abortion<br />msw<br />msw<br />late aborti...
ADV<br />
PPV<br />Inoculated po at 47 days of gestation<br />Euthanized at 81 days of gestation<br />Natural case<br />At birth<br />
Inoculated at 43 days of gestation<br />Euthanized at 98 days of gestation<br />PPV<br />
Virus<br />+<br />mummified<br />+<br />+<br />autolytic<br />normal<br />PCV2<br />Aspect<br />at birth<br />Inoculated <...
mummified<br />Myocarditis<br />PCV2<br />inoculated at 57 days of gestation<br />collected at 21 days PI<br />Ascites,<br...
PRRSV<br />
PEV<br />Inoculated in fetus 8  <br />at 59 days of gestation<br />Hysterectomy at 113 days<br />of gestation<br />7 livin...
CSFV<br />Inoculation of CSFV (low virulence) po at 43 days of gestation<br />Collection at parturition<br />
Reproductive failure - Diagnosis?<br />Main complaint: abortion ≠ stages of gestation<br />Fetuses<br />fresh, rigor morti...
Reproductive failure - Diagnosis?<br />Main complaint: SMEDI<br />Fetuses/piglets at birth<br />Mummies (≠ stages), stillb...
Reproductive failure - Diagnosis?<br />Main complaint: late abortion/early farrowing<br />Fetuses at late abortion/early f...
Control<br />1° Eradication<br />ADV √<br />CSFV √<br />Other viruses in the future?      PRRSV?<br />
Control<br />2° Vaccination/”controlled” immunisation<br />PPV<br />PCV2<br />PEV<br />PRRSV<br />
Gilts<br />OK<br />PPV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br /...
Gilts<br />PPV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 kg...
Gilts<br />Gilts<br />V<br />V<br />V<br />PPV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br />...
Gilts<br />PCV2<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br ...
Gilts<br />PCV2<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 k...
Gilts<br />Gilts<br />V<br />V<br />PCV2<br />V<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br /...
Gilts<br />PRRSV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br...
Gilts<br />PRRSV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 ...
Gilts<br />Gilts<br />V<br />V<br />PRRSV<br />V<br />OK?<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br...
Gilts<br />PRRSV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />!<br />Sows +<br />offspring<br /...
Gilts<br />PEV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />20-40 kg<br />
Laboratory of Virology<br />& Laboratory of ReproductiveTechnology<br />Faculty of VeterinaryMedicine<br />GhentUniversity...
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01 hans nauwynck

  1. 1. Viral reproductive disorders in sows<br />an update 2011<br />H. Nauwynck<br />Laboratory of Virology<br />Faculty of Veterinary Medicine<br />Ghent University, Belgium<br />
  2. 2. 1. CL (progesterone)<br />Normalgestation in the sow<br />2. Cytokine/growth factor <br />cross talk<br />?<br />3. Immune suppression<br />
  3. 3. How do viruses cause reproductive problems?<br />General disease (fever) <br /> ~ pro-inflammatory cytokines causing distortion of cytokine balance<br /> -> swine influenza virus, …<br />2. Replication of pathogens in reproductive tract/embryo/fetus <br /> ~ damaging the placenta and/or cytokine balance<br /> ~ direct negative impact on embryo/fetus<br />-> DNA viruses: Aujeszky’s disease virus, parvovirus, porcine circovirus 2<br />-> RNA viruses: porcine reproductive and respiratory syndrome virus, porcine enteroviruses, encephalomyocarditis virus, classical swine fever virus<br />
  4. 4. How do virusesreachembryos/fetuses? <br />1. Contaminatedsperm<br />
  5. 5. 2. Viremia + transplacental spread<br />How do virusesreachembryos/fetuses? <br />
  6. 6. 1. Via contaminatedsperm<br />Virus excretion in sperm<br />~ viremia<br />DNA-viruses<br />ADV (<14 dPI)<br />PPV (<14 dPI)<br />PCV2 (<50 dPI;>)<br />RNA-viruses<br />PRRSV (<50 dPI;>)<br />PEV (<14 dPI)<br />CSFV (<14 dPI)<br />Embryo<br />Virus rep & †<br />Virus rep & †<br />Virus rep & †<br />No virus rep (onlyinfectionsow) <br />Virus rep & †<br />Virus rep & †<br />repeat breeding<br />repeat breeding<br />
  7. 7. At what stage can embryos be infected ?<br /> 2-8 cells morula blastocyst hatched blastocyst<br />(0-4dpins) (4-5dpins) (5-7dpins) (>7dpins)<br />-<br />-<br />-<br />-<br />?<br />?<br />-<br />-<br />-<br />-<br />?<br />?<br />-<br />-<br />-<br />-<br />?<br />?<br />+<br />+<br />+<br />-<br />+<br />+<br />ADV<br />PPV<br />PCV2<br />PRRSV<br />PEV<br />CSFV<br />Virus should resist aggressive uterine environment!!!<br />1° difficult to inactivate: PPV, PCV2, PEV (no envelope)<br />2° intracellular: ADV, CSFV<br />
  8. 8. Zona pellucida is an ideal barrier<br />1° Physical barrier<br />Channels in ZP : ± 30 nm (inner ø)<br />Viruses > 30nm (ADV, PRRSV, CSFV) <br />2° Chemical barrier<br />Viruses < 30nm (PPV, PCV2, PEV)<br />
  9. 9. Outcome of embryonic PPV-infection<br />Uninfected<br />PPV-infected<br />
  10. 10. Outcome of embryonic PCV2-infection <br />Surgical ET<br />HB 6 dpins<br />PCV2/mock<br />Inspection of <br />ovaries<br />(follicles/CL)<br />Euthanasia<br />D14 post ET<br />Collection & analysis of embryos<br />
  11. 11. Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />at transfer<br />n° status<br />26 mock<br />26 mock<br />20 PCV2<br />23 PCV2<br />20 PCV2<br />18 PCV2<br />29 PCV2<br />Ovaries<br />CL<br />CL<br />CL<br />CL<br />follicles<br />follicles<br />follicles<br />Embryos<br />at euthanasia<br />n°<br />19<br />17<br />13<br />3<br />0<br />0<br />0<br />normal<br />19<br />15<br />7<br />0<br />0<br />0<br />0<br />degenerated<br />0<br />2<br />6<br />3<br />0<br />0<br />0<br />mock<br />normal<br />embryo<br />PCV2<br />degenerated<br />embryo<br />
  12. 12. Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />at transfer<br />n° status<br />26 mock<br />26 mock<br />20 PCV2<br />23 PCV2<br />20 PCV2<br />18 PCV2<br />29 PCV2<br />Ovaries<br />CL<br />CL<br />CL<br />CL<br />follicles<br />follicles<br />follicles<br />Embryos<br />at euthanasia<br />n°<br />19<br />17<br />13<br />3<br />0<br />0<br />0<br />normal<br />19<br />15<br />7<br />0<br />0<br />0<br />0<br />degenerated<br />0<br />2<br />6<br />3<br />0<br />0<br />0<br />0<br />0<br />0<br />4<br />6<br />3<br />IHC-PCV2<br />
  13. 13. Outcome of embryonic PCV2-infection <br />Sow<br />n°<br />A<br />B<br />C<br />D<br />E<br />F<br />G<br />Embryos<br />at transfer<br />n° status<br />26 mock<br />26 mock<br />20 PCV2<br />23 PCV2<br />20 PCV2<br />18 PCV2<br />29 PCV2<br />Embryos<br />at euthanasia<br />Ovaries<br />CL<br />CL<br />CL<br />CL<br />follicles<br />follicles<br />follicles<br />n°<br />19<br />17<br />13<br />3<br />0<br />0<br />0<br />normal<br />19<br />15<br />7<br />0<br />0<br />0<br />0<br />degenerated<br />0<br />2<br />6<br />3<br />0<br />0<br />0<br />0<br />0<br />0<br />4<br />6<br />3<br />PCV2 infection of embryos leads to<br />• embryonic death<br />• return to oestrus of sow (regular/irregular)<br />• both PCV2a and PCV2b<br />IHC-PCV2<br />
  14. 14. 2. Viremia + transplacental spread<br />DNA-viruses<br />(PPV, PCV2, ADV)<br />RNA-viruses<br />(PRRSV, CSFV, <br />PEV 1, 3, 6 & 8, EMCV)<br />• Cell-free: PPV, PEV, PRRSV, EMCV<br />• Cell-associated: ADV, PCV2, CSFV<br />
  15. 15. Transplacental spread - cell free virus<br />PRRSV (> 70dpi)<br />PPV, PEV, EMCV<br />*<br />*<br />*<br />*<br />*<br />*<br />
  16. 16. Normal placenta<br />fetal site<br />maternal site<br />
  17. 17. Transplacentalspread- PRRSV - 1. cell free virus + free mø<br />
  18. 18. Transplacental spread - PRRSV - 2. cell free virus + migrating mø<br />
  19. 19. Transplacental spread - PRRSV - 3. infection of migrating mø<br />
  20. 20. Transplacental spread - PRRSV - 4. entry of fetal placenta by <br />migrating mø<br />
  21. 21. Transplacental spread - PRRSV - 5. explosive replication in fetal placental mø <br />
  22. 22. Transplacental spread - PRRSV - 6. apoptosis in infected mø and bystander cells<br />
  23. 23. Transplacental spread - PRRSV - 7. apoptosis in infected mø and bystander cells/necrosis?<br />
  24. 24. Transplacental spread - PRRSV - 8. dysfunction placenta<br />
  25. 25. Transplacental<br />spread-<br />cell-associated virus<br />*<br />*<br />*<br />*<br />PCV2, ADV, CSFV <br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />*<br />
  26. 26. Cell-associated transplacental spread of ADV<br />Maternal<br />placenta<br />Fetal<br />placenta<br />Maternal<br />placenta<br />
  27. 27. Interfetal virus spread<br />PPV/PEV/PRRSV<br />ADV/CSFV<br />PCV2<br />*<br />
  28. 28. Clinical outcome - fetus <br />Infection time (days of gestation)<br />0-30<br />†<br />†<br />†<br />-<br />†<br />†<br />30-60<br />†<br />†<br />†<br />(†)<br />†<br />†/IT<br />terato<br />60-90<br />†<br />†/IR<br />†/IR<br />†<br />†/IR<br />†/IR<br />90-113<br />†<br />IR<br />IR<br />†/IR<br />IR<br />†/IR<br />ADV<br />PPV<br />PCV2<br />PRRSV<br />PEV<br />CSFV<br />IR immune response<br />IT immune tolerance<br />
  29. 29. Clinical outcome - sow<br />Infection time (days of gestation)<br />60-90<br />abortion<br />msw<br />msw<br />late abortion/msw<br />msw<br />abortion/msw<br />90-113<br />abortion/msw<br />normal<br />normal<br />late abortion/msw<br />normal<br />abortion/msw<br />0-30<br />repeat br.<br />repeat br.<br />repeat br.<br />-<br />repeat br.<br />repeat br.<br />30-60<br />abortion<br />msw<br />msw<br />-<br />msw<br />abortion/msw<br />ADV<br />PPV<br />PCV2<br />PRRSV<br />PEV<br />CSFV<br />msw: mummies, stillborn and weakborn piglets at birth <br />
  30. 30. ADV<br />
  31. 31. PPV<br />Inoculated po at 47 days of gestation<br />Euthanized at 81 days of gestation<br />Natural case<br />At birth<br />
  32. 32. Inoculated at 43 days of gestation<br />Euthanized at 98 days of gestation<br />PPV<br />
  33. 33. Virus<br />+<br />mummified<br />+<br />+<br />autolytic<br />normal<br />PCV2<br />Aspect<br />at birth<br />Inoculated <br />(days of gestation)<br />Intra-uterine<br />spread<br />Interruption<br />of gestation<br />Abs<br />57<br />mummified<br />-<br />+<br />-<br />stillborn<br />autolytic<br />75<br />+<br />-<br />-<br />92<br />normal<br />+<br />-<br />-<br />
  34. 34. mummified<br />Myocarditis<br />PCV2<br />inoculated at 57 days of gestation<br />collected at 21 days PI<br />Ascites,<br />congestion <br />of internal organs<br />At birth<br />
  35. 35. PRRSV<br />
  36. 36. PEV<br />Inoculated in fetus 8 <br />at 59 days of gestation<br />Hysterectomy at 113 days<br />of gestation<br />7 living fetuses<br />
  37. 37. CSFV<br />Inoculation of CSFV (low virulence) po at 43 days of gestation<br />Collection at parturition<br />
  38. 38. Reproductive failure - Diagnosis?<br />Main complaint: abortion ≠ stages of gestation<br />Fetuses<br />fresh, rigor mortis (firm)<br />Fetuses<br />not fresh, partially mummified<br />Lungs/spleen fetuses<br />(esp. with necrotic foci in liver)<br />No serum sows!!!<br />Immunofluorescence<br />Virus isolation/PCR<br />ADV, CSFV <br />Sera sow<br />Seroconversion<br />SIV or other pathogen<br />causing general disease<br />fever <br />
  39. 39. Reproductive failure - Diagnosis?<br />Main complaint: SMEDI<br />Fetuses/piglets at birth<br />Mummies (≠ stages), stillborn and normal piglets <br />1) Heart/spleen/lungs fetuses (<70 dpins; <17cm)<br />2) Body fluid/serum fetuses (>70dpins; >17cm)<br />No serum sows!!!<br />1) Virus isolation/PCR<br />2) Antibody detection (BE CAREFULL WITH PCV2!!!)<br />PPV, PCV2, PEV, EMCV<br />
  40. 40. Reproductive failure - Diagnosis?<br />Main complaint: late abortion/early farrowing<br />Fetuses at late abortion/early farrowing<br />Early stages of mummification (brown), stillborn and normal piglets <br />Placenta/lungs/spleen fetuses<br />Serum stillborn/piglets before colostrum uptake<br />No serum sows!!!<br />Virus isolation(±)/PCR<br />PRRSV<br />
  41. 41. Control<br />1° Eradication<br />ADV √<br />CSFV √<br />Other viruses in the future? PRRSV?<br />
  42. 42. Control<br />2° Vaccination/”controlled” immunisation<br />PPV<br />PCV2<br />PEV<br />PRRSV<br />
  43. 43. Gilts<br />OK<br />PPV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br />OK<br />
  44. 44. Gilts<br />PPV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 kg<br />
  45. 45. Gilts<br />Gilts<br />V<br />V<br />V<br />PPV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br />Sows +<br />offspring<br />20-40 kg<br />V<br />
  46. 46. Gilts<br />PCV2<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br />OK<br />
  47. 47. Gilts<br />PCV2<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 kg<br />
  48. 48. Gilts<br />Gilts<br />V<br />V<br />PCV2<br />V<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br />Sows +<br />offspring<br />20-40 kg<br />V<br />
  49. 49. Gilts<br />PRRSV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />OK<br />20-40 kg<br />OK<br />
  50. 50. Gilts<br />PRRSV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 kg<br />
  51. 51. Gilts<br />Gilts<br />V<br />V<br />PRRSV<br />V<br />OK?<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />OK<br />OK<br />Sows +<br />offspring<br />20-40 kg<br />V<br />
  52. 52. Gilts<br />PRRSV<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />!<br />!<br />!<br />!<br />Sows +<br />offspring<br />20-40 kg<br />
  53. 53. Gilts<br />PEV<br />OK<br />√<br />40-100 kg<br />10-20 kg<br />Sows<br />Sows +<br />offspring<br />20-40 kg<br />
  54. 54. Laboratory of Virology<br />& Laboratory of ReproductiveTechnology<br />Faculty of VeterinaryMedicine<br />GhentUniversity, Belgium<br />H. Nauwynck<br />Van Soom<br />G. Labarque<br />R. Sanchez<br />G. Vanroose<br />S. Tanghe<br />B. Mateusen<br />P. Meerts<br />D. Lefebvre<br />D. Saha<br />U. Karniychuk<br />E. Bussalleu<br />J. Beek<br />Virus-embryo/fetus work<br />

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