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02 pcvd g sarli

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  • 1. 4TH MERIAL FORUM HAVE WE GOT PCVD &SWINE INFLUENZA UNDER CONTROL?
  • 2. PCV2 impact and control on early stage of gestation in sows Giuseppe Sarli 4rd Merial Forum – Berlin – May 31st 2012
  • 3. Target …..• Results of a study conducted in 2010EXPERIMENTAL INFECTION WITH PCV2 EXPOSED SEMEN OF CONVENTIONAL CIRCOVAC® VACCINATED AND NON- VACCINATED SOWS • AASV Meeting 2012
  • 4. Objectives of the studyTo evaluate viremia, virusshedding and viral load inmaternal serum, tissuesand fetuses ina.6 PCV2 infectedCircovac© vaccinated giltsb.6 PCV2 infected giltsc.3 Control gilts
  • 5. Trial 2009 ……..PCV2 impact on early stage ofgestation in sows: experimentalresults• 6 PCV2 infected gilts• 3 Control gilts
  • 6. Trial 2009 results …..• 4 out of the 6 infected sows developed viremia • Viremia was longer in the sow with the lowest anti- PCV2 antibody titre• In at least one organ of the six infected sows PCV2 was detected • The sow with the lowest anti-PCV2 antibody titre showed the higher number of positive tissues and fetuses
  • 7. Groups for trial 2010 • 6 Circovac® + PCV2bHormonal estrus synchronization infected gilts (VI) – First vac.: 16 weeks Artificial insemination with – Second vac.: 20 weeks PCV2b spiked semen = 4x1010 genome copy/ml = 10 3.9 TCID50/ml • 6 PCV2b infected gilts (NVI) Artificial insemination with • 3 Controls (C) PCV2 negative semen
  • 8. Experimental design Control PCV2 PCV2 infected infected + Circovac ® Samples analysisD0 PCV2 PCV2  semen Cervix, nasal, rectal swabs and bloodAI semen 10 ml 10 ml 103.9TCID50/ml sample every weekD29 Ultrasonography Sow: uterus, lymphD30 nodes placenta; Euthanasia of empty sows/D47 Fetus: lung, amniotic fluid, heart, liverD52 RtPCR Euthanasia of pregnant sows/D56 Histology, IHC
  • 9. Material and Methods• Serology• Real time PCR – Swabs (vaginal, rectal, nasal) – Blood – Foetuses – Gilts tissues• Histology – H&E – Immunohistochemistry (for tissue ≥ 108 PCV2 genome copies/ml)
  • 10. B: bloodSampling S: swabs : day of euthanasia
  • 11. Pregnancy• VI – 3 out of 6 gilts were pregnant• NVI – 3 out of 6 gilts were pregnant• C – 2 out of 3 gilts were pregnant
  • 12. Serology: antibody titre
  • 13. Real time PCR: serum, swabs and fetuses ID Sera Fecal swabs Vaginal Nasal swabs gilts swabs 108 ooo oo oooo 109 oo oo oo 110 o o oo IV 112 o o o 114 oo ooo 115 o ooo o o 117 ooo oooooo ooooo ooooo 118 ooooo oooooo ooooo oooooo 119 oooooooo ooooooooo oooooo oooooooo INV 123 oo ooooooo ooo ooo 126 oo oooo oooo ooo 127 o oooooo oo oooo 116 o oooo oooo ooo CTR 120 oo oooooooo ooo oooo 122 oooooo oooooooo oooooooo ooooooooo
  • 14. Real time PCR: serum, swabs and fetuses n°positive samples n° sampling for each animal N° positive N° positive N° positive N° positive N° positive N° positiveGroup ID N° viremia vaginal nasal amniotic fecal swabs tissue fetus swabs swabs fluid 108 0/8 2/9 4/9 3/9 12/20 0/3 0/3 Not Not 109 0/5 2/6 2/6 2/6 15/15 pregnant pregnant Aborted Aborted 110 1/4 0/5 2/5 1/5 15/15 IV PCR- PCR- Aborted Aborted 112 0/4 1/5 1/5 1/5 13/15 PCR- PCR- 114 0/7 0/8 3/8 2/8 23/42 0/15 0/15 115 1/8 1/9 1/9 3/9 5/30 0/9 0/9 total 2/36 6/42 13/42 12/42 83/137 0/27 0/27 Not Not 117 3/5 5/6 5/6 6/6 15/15 pregnant pregnant Not Not 118 5/5 5/6 6/6 6/6 15/15 pregnant pregnant INV 119 8/8 6/9 8/9 9/9 31/32 1/10 2/10* 123 2/7 3/8 3/8 7/8 42/46 0/16 1/16 Aborted Aborted 126 2/4 4/5 3/5 4/5 15/15 PCR- PCR- 127 1/8 2/9 4/9 6/9 36/41 0/14 0/14 total 21/37 25/43 29/43 38/43 153/164 1/40 3/40 Not Not 116 1/5 4/6 3/6 4/6 15/15 pregnant pregnantCTR 120 1/8 3/9 4/9 8/9 26/26 0/7 0/7 122 6/8 8/9 9/9 9/9 28/32 1/10 0/10 total 8/21 15/24 16/24 20/24 69/73 1/17 0/17
  • 15. IV INV CTR Non- Non-pregnant Pregnant Pregnant sows Non-pregnant sows Pregnant sows pregnant sows sows sowsSows ID 108 114 115 109 110 112 119 123 127 117 118 126 120 122 116Number offetuses 3 15 9 0 0 0 10 16 14 0 0 0 7 10 0Left tonsil + + - + + + + + + + + + + + +Right tonsil - - - + + + + + + + + + + + +Left uteriunelymph node - + - + + + + + + + + + + + +Right uteriunelymph node + - - + + - + + + + + + + + +Left inguinallymph node - - - + + + + + + + + + + + +Right inguinallymph node + - - + + - + + + + + + + + +Left ovary + + - + + + + + + + + + + + +Right ovary + - - + + + + + + + + + + + +Left salpinx + + - + + + + + + + + + + + +Right salpinx - - - + + + + + + + + + + + +Vagina + + + + + + + + + + + + + + +Cervix - - + + + + + + + + + + + + +Fetuses (pos/tot.) - - - / / / 1/10 - - / / / - 1/10 /Amniotic fluid - = 40% - - / / / 2/10 1/16 - / / / = 76% - - /(pos/tot.) = 77%Placenta(pos/tot.) 2/3 8/15 1/9 / / / 9/10 14/16 8/14 / / / 7/7 6/10 /Left uterin tracts* + + + + + + + + + + + + + + +Right uterintracts* + + + + + + + + + + + + + + +
  • 16. Real time PCR: tissues N° PCV2 positive N° positive tissue ≥ 108 PCV2 genome Group Total tissue valued tissues copies/ml IV 127 83 6 INV 164 153 42 CTR 73 69 17
  • 17. HistologyAnimal (ear tag and group) Tissue PCV2 antigen localization Follicular dendritic cell 117 INV Left tonsil occasional histiocytic Follicular dendritic cell 117 INV Right tonsil occasional histiocytic Follicular dendritic cell 118 INV Left tonsil occasional histiocytic Follicular dendritic cell 118 INV Right tonsil occasional histiocytic Left superficial inguinal lymph 118 INV Rare histiocytic node Right superficial inguinal 118 INV Rare histiocytic lymph node
  • 18. Summarizing the results …..The vaccine was shown to have an active role inreducing virus shedding and tissues viral load:1.lower number of positive in vivocollected samples in the VI groupcompared to NVI and C
  • 19. Summarizing the results …..The vaccine was shown to have an active role inreducing virus shedding and tissues viral load:2.lower percentage of placentas and nopositive fetuses in VI gilts compared toNVI and C groups
  • 20. Summarizing the results …..• Considering a similar antibody level at the beginning of the experiment, their quantitation should not take into account the whole vaccine effect.• Role for mucosal immunity triggered by vaccination ? Duodenum+jejunum+ileum
  • 21. Acknowledgments• Dept. Veterinary Medical • IZSLER Science – Bologna – Ostanello Fabio Infectious diseases LS – Dottori Michele Reggio – Morandi Federico – Bonilauri Paolo Emilia – Bianco Carlo – Cordioli Paolo – Panarese Serena Pathology Brescia LS – Lelli Davide – Felice Viviana – Ferrara Domenico – Bacci Maria Laura • Merial – Govoni Nadia Physiology LS – Joisel Francois – Galeati Giovanna Lyon – Thais Vila Italy – Giorgio Leotti
  • 22. Thank you for your attention
  • 23. Giuseppe SarliDepartment of Veterinary Medical Science giuseppe.sarli@unibo.it www.unibo.it

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