PCV2: Diagnostics, Control, Protection, and Efficacy Measures - Dr. Brian Payne, Boehringer Ingelheim Vetmedica, Inc, from the Boehringer Ingelheim Pre-AASV Conference, February 28, 2014 - Dallas, TX More presentations at http://www.swinecast.com/2014-boehringer-ingelheim-aasv

1. PCV2: Diagnostics, Control,
Protection, and Efficacy Measures
Brian Payne, DVM
Boehringer Ingelheim Vetmedica, Inc, St. Joseph, MO,
USA

2. Crash course into PCV2
• PCV2 timeline
• Case load over time
• Diagnostics
• Duration of immunity
• Cross-protection and Mutations
• Vaccine measurements

3. 40 years of PCV2 (select events)
•PCV2 discovered in kidney cell (PK15) line (1974, Tischer)
•PCV and congenital tremors (1994, Hines and Lukert)
•PCV and PMWS linked (1997, Clark)
• First PCV pathogenesis study (1986, Tischer)
•Etiologic agent and PMWS described (1998, Allan, Ellis, Meehan)
FLEXcombo®
3FLEX®
*FLEXcombo’s predecessor was Ingelvac CircoFLEX® – MycoFLEX®
CircoFLEX®
MycoFLEX®
•Classification of PCV1 and PCV2 (1998, Meehan)
Clusters PCV2a, PCV2b nomenclature (2007-8, Olvera, Segalés) ●
PCVAD described (2007, Opriessnig) ●
•PDNS described (2001, Choi)
12-59% morbidity rates in unvaccinated (04-07, USDA)●
Horizontal transmission described (03-11 multiple authors) ●
•Vertical transmission described (99-00, multiple authors)
Global shift from PCV2aPCV2b described (07-08 multiple authors)●
PCV2 vaccines in N. America (2006) ●
First PCV2 vaccine launched in EU (2004, Reynaud) ●
AASV taskforce (06-09) ●
20141974 1994 20041984 1999 2009
PCV2 clinical sign nomenclature (2012, Segalés) ●
•Serology survey throughout the globe (1982-95, Tischer, Dulac, Harding, Clark, Horner, Edwards, Hines)

4. PCV2 Cases overtime – Iowa State U.
Opriessnig, 2007, J Vet Diagn invest
Overall trend of porcine circovirus type 2–associated disease (PCVAD) cases submitted to the Veterinary Diagnostic
Laboratory at Iowa State University.

5. PCV2 Cases overtime – Iowa State U.
Opriessnig, 2011, Animal Health Research Reviews
# cases
Madson, ISU-VDL, January 2014

6. Routine Diagnostics
Serum
Tissues
PUCS
Oral fluids
Environmental
PCR
ELISA
Sequencing
Nucleotide Substitution per 100 residues
0
7.1
246
13-49311-14 (BIVI 129804)
13-49311-23 (BIVI 129804)
13-49313-12 (BIVI 131678)
13-49314-504 (BIVI 131263)
13-49314-506 (BIVI 131263)
13-49317-426 (BIVI 130279)
Histopathology
IHC

7. Updated PCV2 qPCR
Chittick, Leman 2013
420
148
PVC2 qPCRs positives
4 recent trials
2-4 logs
>4 logs

8. Ingelvac CircoFLEX® PCV2 protection

9. Parma Ham Studies
Terreni, ESPHM 2009a Terreni, ESPHM 2009b Persico, ESPHM 2013
285 days on feed
CircoFLEX®
303 days on feed
CircoFLEX®
265 days on feed
FLEXcombo®
http://www.epicurus.com

10. Iberian Ham Studies
Palomo, IPVS 2012 Criado, Emerging & Remerging Diseases, 2011
10 months old
FLEXcombo®
10 months of age
FLEXcombo®
http://www.ibergour.co.uk/

11. US Field Studies
3 replicates
2,338 pigs
1.45
1.50
1.55
1.60
1.65
1.70
1.75
d0-first cut d0-second cut d0-final cut
ADG,pounds,lbs
ADG – competitive PCV2/M. hyo
vaccine study
FLEXcombo® Circumvent PCVM®
a
a
ab
b
b
ab Means differ (Tukey’s HSD P≤0.05)
BIVI 2011093; Payne IPVS 2012
100% of the pigs,measured all the way to market
0
10
20
30
40
50
60
70
80
90
100
1 35 70 105 139
PCV2QPCRPercent>4Logs
Day of Trial
PCV2 qPCR % Positive

12. Yes, PCV2 mutants exist
Takahagi, Differential Effects of Porcine
Circovirus Type 2 (PCV2) Vaccination on
PCV2 Genotypes at Japanese Pig Farms,
Virology, 2009, 35-41.
Opriessnig, Halbur, ISU Dz Conf, 2013, p30-34.

13. CircoFLEX® cross-protection papers
• Licensing Data*
• Takahagi, 2009, 16 strains cross-protection paper
• Derosiers, JSHAP, 2009
• Cline, Vet Record, 2008
• Haiwick, AASV, 2014
• Werling, AASV, 2014
*BIVI 6131-0981-04P-036

14. CircoFLEX® Cross-protection papers
Takahagi, Differential Effects of Porcine Circovirus Type 2 (PCV2) Vaccination
on PCV2 Genotypes at Japanese Pig Farms, Virology, 2009, 35-41.
Viremia reduced in all cases

15. Haiwick, AASV 2014 Oral Presentation
Note: No statistics were conducted on this data

16. Werling, 2014 AASV, poster
• Designed to evaluate CMI
* = p<0.05;
** = p,0.01;
*** = p<0.001
** *
***
***
** *

17. Novel mutations of PCV2

18. Novel (2012) PCV2b described by Dr. Opriessnig
Xiao, Opriessnig, JVI, 2012
Opriessnig, AASV, 2012
Opriessnig, Vet Micro 2012 paper
ISU diagnostic reports

19. PCV2a PCV2b “mPCV2b”
ORF 1 PCR + + +
ORF 2 a/b
differential
+ (a) + (b) +(b) / -
# nucleotides 702 702 705
# amino acids 233 233 234
Comparison between PCV2 strains
Opriessnig, Halbur, ISU Dz Conf, 2013, p30-34
• ORF 2 homology to:
• PCV2a: 90.6-91.0%
• PCV2b: 93.6-94.9%
• ORF1 homology to:
• PCV2a: 99.0-99.4%
• PCV2b: 98.7%

20. Measurements
• Clinical picture
• Diagnostics
• Performance

21. Pre-PCV2 vaccines
• Prior to commercially available PCV2 vaccines,
several PCV2 studies demonstrated that levels of
PCV2 virus in the serum (viremia) and tissues
were correlated to the severity of disease1-12
• Clinically ill pigs had higher viremia and PCV2 tissue levels
than subclinical pigs
• In these non-vaccinated pigs, lesions correlated
with viral load
• Suggestions that 107 viral load could be used to
diagnose PMWS1-3

22. PCV2 vaccines enter the market
• In vaccinated pigs high levels of viremia are rarely seen13
• All commercial vaccines reduce viremia16,17
• In vaccinated pigs, viremia is not predictive to production
performance
• North American14 and European15 studies
• Different PCV2 protocols (1 vs. 2 dose) show no
difference in magnitude of viremia13
• Elimination of PCV2 with vaccine has NOT been
demonstrated

23. Holck, Leman 2009
Maass, Leman 2009
Viremia and ADG – no correlation

24. Percentage of pigs positive, PCV2
qPCR
TRIALATRIALB
0%
20%
40%
60%
80%
100%
3 10 13 16 19 22
Weeks of Age
NVC
FLEX
PCV M
0%
20%
40%
60%
80%
3 7 10 14 19 23
NVC
CircoFLEX
Fostera
Bretey et al. 2013. Leman Conference

25. n
Start
Avg. wt
Start
Tot Wt
Start, lbs
n
End
Avg. wt
End, lbs
Total Wt
End, lbs
Avg. wt
Gain, lbs
Circo
FLEX®
Fostera®
PCV
Circo
FLEX®
Fostera®
PCV
Circo
FLEX®
Fostera®
PCV
n
Start
444 444 145 150 433 431
Avg. wt
Start
13.98 14.12 14.19 14.22 18.75 18.64
Tot Wt
Start, lbs
6,207.12 6,269.28 2,057.99 2,133.45 8,118.75 8,033.84
n
End
425 421 144 149 413 410
Avg. wt
End, lbs
243.60 242.19 260.21 256.51 237.72 236.28
Total Wt
End, lbs
103,530 101,962 37,470.2 38,220.0 98,178.4 96,874.8
Avg. wt
Gain, lbs
229.62 228.07 246.021 242.288 218.97 217.64
Circo
FLEX®
Fostera®
PCV
1,022 1,025
16.03 16.03
16,383.9 16,436.6
982 980
243.61 241.99
239,178.6 237,056.8
227.58 225.96
3 Trial Results
↓3 pigs
Equal
↑2 pigs
↑1.62 lbs
↓52.70 lbs
↑2,121.8 lbs
↑1.62 lbs
Trial 1* Trial 2* Trial 3* Combined
Notethatthesearenumericaldifferences;*BIVI2011191; BIVI2012056; BIVI2011252

26. Example scenario financials
CircoFLEX® Vaccine B
Number of pigs started 1,000 1,000
Starting Weight 16.0 lbs
Mortality percent 3.9% 4.4%
Total pigs marketed 961 956
Days on Feed 140 days
ADG, lbs 1.63 1.62
Average live weight per pig, lbs 244.2 lbs 242.8 lbs
Carcass yield estimate 74%
Estimated carcass weight 180.7 179.7
Total wt. of carcasses sold 173,653 lbs 171,793 lbs
Difference in lbs, kg 1,860 lbs = 844 kg
Carcass price , Dec 2013* $0.76 / lb
Diff. in total carcass price $1,414
Total carcass price/1,000 pigs $1.41/pig benefit CircoFLEX® over Vaccine B
* http://www.agmanager.info/livestock/marketing/graphs

27. FLEX Family
Customized Control
because every herd is unique