02 pcvd g sarli

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


pregnant

• C


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


118 INV Left tonsil Follicular dendritic cell


118 INV Right tonsil Follicular dendritic cell


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


• 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

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

[email protected]

www.unibo.it

mailto:[email protected]

http://www.unibo.it/

02 pcvd g sarli

Business

Transcript of 02 pcvd g sarli