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4TH MERIAL FORUM
HAVE WE GOT PCVD &
SWINE INFLUENZA UNDER
CONTROL?
PCV2 impact and control on early
stage of gestation in sows
Giuseppe Sarli
4rd Merial Forum – Berlin – May 31st 2012
Target …..
• Results of a study conducted in 2010
EXPERIMENTAL INFECTION WITH PCV2
EXPOSED SEMEN OF CONVENTIONAL
CIRCOVAC® VACCINATED AND NON-
VACCINATED SOWS
• AASV Meeting 2012
Objectives of the study
To evaluate viremia, virus
shedding and viral load in
maternal serum, tissues
and fetuses in
a.6 PCV2 infected
Circovac© vaccinated gilts
b.6 PCV2 infected gilts
c.3 Control gilts
Trial 2009 ……..
PCV2 impact on early stage of
gestation in sows: experimental
results
• 6 PCV2 infected gilts
• 3 Control gilts
• 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
Trial 2009 results …..
Groups for trial 2010
• 6 Circovac® + PCV2b
infected gilts (VI) – First vac.: 16 weeks
– Second vac.: 20 weeks
• 6 PCV2b infected gilts
(NVI)
• 3 Controls (C)
Artificial insemination with
PCV2b spiked semen
= 4x1010 genome copy/ml
= 10 3.9 TCID50/ml
Artificial insemination with
PCV2 negative semen
Ho
rmo
nal estr
us s
yn
ch
ron
izati
on
Experimental design Control PCV2
infected PCV2 infected + Circovac ®
Samples analysis
D0 AI
PCV2 semen
10 ml
PCV2 semen 10 ml
103.9TCID50/ml
Cervix, nasal, rectal swabs and blood sample every week
D29 Ultrasonography
D30 /D47
Euthanasia of empty sows
Sow: uterus, lymph nodes placenta; Fetus: lung, amniotic fluid, heart, liver
RtPCR Histology, IHC
D52 /D56
Euthanasia of pregnant sows
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)
Sampling B: blood
S: swabs
: day of euthanasia
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
Serology: antibody titre
Real time PCR: serum, swabs and fetuses
ID
gilts
Sera Fecal swabs Vaginal
swabs
Nasal swabs
IV
108 ooo oo oooo
109 oo oo oo
110 o o oo
112 o o o
114 oo ooo
115 o ooo o o
INV
117 ooo oooooo ooooo ooooo
118 ooooo oooooo ooooo oooooo
119 oooooooo ooooooooo oooooo oooooooo
123 oo ooooooo ooo ooo
126 oo oooo oooo ooo
127 o oooooo oo oooo
CTR
116 o oooo oooo ooo
120 oo oooooooo ooo oooo
122 oooooo oooooooo oooooooo ooooooooo
Group ID N° viremia N° positive
vaginal swabs
N° positive nasal swabs
N° positive fecal swabs
N° positive tissue
N° positive fetus
N° positive amniotic
fluid
IV
108 0/8 2/9 4/9 3/9 12/20 0/3 0/3
109 0/5 2/6 2/6 2/6 15/15 Not
pregnant Not
pregnant
110 1/4 0/5 2/5 1/5 15/15 Aborted
PCR- Aborted
PCR-
112 0/4 1/5 1/5 1/5 13/15 Aborted
PCR- Aborted
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
INV
117 3/5 5/6 5/6 6/6 15/15 Not
pregnant Not
pregnant
118 5/5 5/6 6/6 6/6 15/15 Not
pregnant Not
pregnant 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
126 2/4 4/5 3/5 4/5 15/15 Aborted
PCR- Aborted
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
CTR 116 1/5 4/6 3/6 4/6 15/15
Not pregnant
Not pregnant
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
Real time PCR: serum, swabs and fetuses n°positive samples n° sampling for each animal
IV INV CTR
Pregnant sows Non-pregnant sows Pregnant sows Non-pregnant
sows Pregnant
sows
Non-pregnant
sows Sows ID 108 114 115 109 110 112 119 123 127 117 118 126 120 122 116 Number of
fetuses 3 15 9 0 0 0 10 16 14 0 0 0 7 10 0
Left tonsil + + - + + + + + + + + + + + + Right tonsil - - - + + + + + + + + + + + + Left uteriune
lymph node - + - + + + + + + + + + + + +
Right uteriune
lymph node + - - + + - + + + + + + + + +
Left inguinal
lymph node - - - + + + + + + + + + + + +
Right inguinal
lymph node + - - + + - + + + + + + + + +
Left ovary + + - + + + + + + + + + + + + Right ovary + - - + + + + + + + + + + + + Left salpinx + + - + + + + + + + + + + + +
Right salpinx - - - + + + + + + + + + + + +
Vagina + + + + + + + + + + + + + + + Cervix - - + + + + + + + + + + + + +
Fetuses (pos/tot.) - - - / / / 1/10 - - / / / - 1/10 /
Amniotic fluid
(pos/tot.) - - - / / / 2/10 1/16 - / / / - - /
Placenta
(pos/tot.) 2/3 8/15 1/9 / / / 9/10 14/16 8/14 / / / 7/7 6/10 /
Left uterin tracts* + + + + + + + + + + + + + + +
Right uterin
tracts* + + + + + + + + + + + + + + +
= 40% = 77%
= 76%
Group Total tissue valued N° PCV2 positive
tissues
N° positive tissue ≥ 108 PCV2 genome
copies/ml
IV 127 83 6
INV 164 153 42
CTR 73 69 17
Real time PCR: tissues
Histology
Animal (ear tag and group) Tissue PCV2 antigen localization
117 INV Left tonsil Follicular dendritic cell
occasional histiocytic
117 INV Right tonsil Follicular dendritic cell
occasional histiocytic
118 INV Left tonsil Follicular dendritic cell
occasional histiocytic
118 INV Right tonsil Follicular dendritic cell
occasional histiocytic
118 INV Left superficial inguinal lymph
node Rare histiocytic
118 INV Right superficial inguinal
lymph node Rare histiocytic
The vaccine was shown to have an active role in
reducing virus shedding and tissues viral load:
1.lower number of positive in vivo
collected samples in the VI group
compared to NVI and C
Summarizing the results …..
The vaccine was shown to have an active role in
reducing virus shedding and tissues viral load:
2.lower percentage of placentas and no
positive fetuses in VI gilts compared to
NVI and C groups
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
Summarizing the results …..
? Duodenum+jejunum+ileum
Acknowledgments
• Dept. Veterinary Medical Science – Bologna
– Ostanello Fabio
– Morandi Federico
– Bianco Carlo
– Panarese Serena
– Felice Viviana
– Ferrara Domenico
– Bacci Maria Laura
– Govoni Nadia
– Galeati Giovanna
• IZSLER
– Dottori Michele
– Bonilauri Paolo
– Cordioli Paolo
– Lelli Davide
• Merial
– Joisel Francois
– Thais Vila
– Giorgio Leotti
Reggio
Emilia
Brescia
Infectious
diseases LS
Pathology
LS
Physiology
LS
Lyon
Italy
Thank you for your attention
Giuseppe Sarli Department of Veterinary Medical Science
www.unibo.it