=Paper= {{Paper |id=Vol-2030/HAICTA_2017_paper1 |storemode=property |title=Epidemiological Investigation of Pseudorabies in Greece |pdfUrl=https://ceur-ws.org/Vol-2030/HAICTA_2017_paper1.pdf |volume=Vol-2030 |authors=Konstantinos Papageorgiou,Evanthia Petridou,George Filioussis,Alexandros Theodoridis,Ioannis Grivas,Odysseas Moshidis,Spiros Kritas |dblpUrl=https://dblp.org/rec/conf/haicta/PapageorgiouPFT17 }} ==Epidemiological Investigation of Pseudorabies in Greece== https://ceur-ws.org/Vol-2030/HAICTA_2017_paper1.pdf

Epidemiological Investigation of Pseudorabies in Greece

Konstantinos Papageorgiou1, Evanthia Petridou1, Georgios Filioussis1, Alexandros
Theodoridis2, Ioannis Grivas3, Odysseas Moshidis4, Spyridon K. Kritas 1
1
Department of Microbiology and Infectious Diseases; 2 Animal Production Economics; 3
Laboratory of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, School
of Health Sciences, Aristotle University of Thessaloniki; and d Department of Business
Administration, University of Macedonia, Thessaloniki, Greece.

Abstract. Pseudorabies, is an acute, frequently fatal disease, that mainly
affects pigs. Although pseudorabies virus (PRV) has been eradicated from
many European countries, it is still endemic in East and Southeast parts of
Europe. Greece belongs to the countries where the disease is enzootic. In this
study, we investigated the presence of PRV in Greek farms. For that reason,
42 pig farms were selected from the entire Greek territory. Blood samples
from different age groups had been collected from each farm and were tested
by ELISA for the presence of antibodies against wild strains of PRV. The
results of our study showed that 28.6% of the selected farms were positive for
the presence of antibodies against wild-type strains of PRV and that factors
such as the non-implementation of biosafety measures and the high-density of
pig farms in an area may affect the probability of a farm to become PRV
positive.

Keywords: pseudorabies, epidemiology, pigs, eradication

1 Introduction
Aujeszky’s Disease or Pseudorabies, is an acute, frequently fatal disease, that
primary affects pigs and incidentally other domestic and wild animals. The term
“pseudorabies” was used as a result of the disease’s clinical resemblance to rabies.
Aladar Aujeszky, was the Hungarian veterinarian who first described and reproduced
the disease in 1902, providing evidence that the etiologic agent was filterable (e.g not
a bacterium but a virus) (Mettenleiter et al., 2012).
PRV is spread all over the world, in parts of Europe, Southeast Asia and America.
In Europe, PRV has been eradicated in Germany, Cyprus, Austria, Sweden, The
Netherlands, Denmark, Czech Republic, Finland, France, Hungary, Luxemburg,
Belgium, Switzerland, Slovakia and UK as a result of the implementation of
eradication programs, but it is still endemic in East and Southeast of Europe (Hahn et
al., 2010). PRV has also been eradicated from Canada, New Zealand and USA
(MacDiarmid, 2000). Although PRV has been eradicated from many countries
throughout the world, the virus is still endemic in the populations of wild boar (Meng
et al., 2009). Therefore, these populations should be considered as potential PRV

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source of infection for domestic pigs. In countries that are free of PRV, vaccination is
prohibited.
Greece belongs to the countries where the disease is still enzootic. According to an
old serological study in 1969 (Papatsas et al., 1995), 20.8% of the collected samples
of domestic pigs from several regions of Greece were positive to antibodies against
PRV. But at that time, there was no serious and organized pig farming in Greece. In
addition, although two recent Greek studies (Touloudi et al., 2015; Marinou et al.,
2015) evidence the presence of PRV in 32% to 35% in wild boars, there is no recent
data regarding the presence of PRV in the population of Greek domestic pigs. Here,
we conducted an epidemiological study in order to investigate the presence of PRV
in the Greek pig farms.

2 Materials and Methods

The study was carried out in Greece from October 2010 to October 2011. Forty-
two (42) farrow-to-finish (FTF) pig herds were selected from the entire Greek
territory at random, based on geographical criteria, in order to obtain representative
data from the population herds. The current study included FTF herds larger than 100
sows, as that kind of farms most likely reflect the commercial pig industry of the
country. The sample represented more than 10% of the FTF farms. The data
regarding the characteristics of the selected herds are presented in table 1.

Table 1. Characteristics of commercial pig herds in Greece (>100 sows) and herd sampling for
the study

Area Density Number of farms sampled /Number of
(km2) (# farms in territory
Territory
farms/ Herd-size category
1000 km2)
Small Large Total
East Macedonia and Thraki 19,000 1.4 4/16 1/10 5/26 (19.2)
Central and West Macedonia 25,000 1.1 4/18 5/11 9/29 (31.0)

Thessalia 14,000 6.9 4/81 3/16 7/97 (7.2)
Epiros and West Sterea Hellas 15,000 6.1 2/66 6/24 8/90 (8.9)
East Sterea Hellas 20,000 1.9 7/21 4/15 12/36 (30.6)
Peloponnesos & Crete 30,000 0.9 1/25 1/12 2/37 (5.4)
TOTAL 123,000 2.4 22/227 20/88 42/315
(9.7%) (22.7%) (13.3%)

The owners or the veterinarians of the selected farms, were contacted in order to
obtain information about the farms and the characteristics of the area where these
holdings were located. The obtained information pertained to:

2
• herd size, e.g. the number of sows on the premises. Farms with less than or
equal to 300 sows were considered as small, while those with more than 300
sows as large.
• pig herd area density e.g. less dense (<20 farms per 1,000 sq km) or more
dense (≥20 farms per 1,000 km2) areas.
• direct distance from the closest pig farm, e.g. short (<6 km) or longer (≥6
km) distance.
• purchase (or not) of breeding animals (gilts or/and boars) from genetic
companies.
• practicing (or not) of at least monthly quarantine in distant building used
exclusively for the newly purchased breeding animals.
• practicing (or not) of certain hygienic/ biosecurity measures at farm.
• practicing (or not) of all-in, all-out (AIAO) flow in all production stages.
• practicing (or not) of vaccination of sows for PRV.
• presence (or not) of substantial economic problems in the farm that
frequently interfere with routine management.
• presence (or not) of certain systemic clinical manifestations at the time of
sampling.
• production stage at which important clinical manifestations were present (or
not) at the time of sampling (neonatal, nursery, grower and finishing stage).

A minimum of 8 blood samples from each out of five different age groups (i.e. 6-,
8-, 10-, 12- and 22-week old pigs) had been collected from each farm (e.g. 40
samples per farm). The blood samples for each age group were collected from pigs of
different pens and, ideally, of different rooms. Sera were individually tested by anti-
PRV-gB ELISA (IDEXX Laboratories, Westbrook, ME) for the presence of
antibodies against the PRV and by anti-PRV-gΕ ELISA (IDEXX Laboratories,
Westbrook, ME) for the differentiation of antibodies against the wild strains of PRV.

Apparent prevalence of PRV infected farms was estimated as the proportion of
farms rearing at least one pig presenting wild-type PRV antibodies. The association
of the herd and neighborhood characteristics of the farms (predictors) with PRV
status was investigated through the application of univariable logistic regression
models with robust standard errors. The SPSS software (IBM Corp. Released 2011.
IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.) was
used. Multiple Correspondence Analysis (MCA) was also used to investigate the
interaction and interrelations among all elements, aiming at the identification of the
dominant and most substantial tendency in their structure.

3 Results – Discussion

The exposure of the farms to wild-type PRV was 28.6% with most of the positive
holdings located in the region of east Macedonia, Thrace and central-west Macedonia
(table 2).

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Table 2. Exposure of Greek farms to wild-type PRV as detected by ELISA

ELISA- gE PRV antibody positive farms /Number of
farms sampled (%)
Herd-size category
Territory Small Territory Total
East Macedonia and Thrace 1/3 1/2 2/5 (40,0%)
Central and West Macedonia 2/3 2/6 4/9 (44,4%)
Thessaly 1/3 1/4 2/7 (28,6%)
Epirus and West Sterea Hellas 1/2 2/6 3/8 (37,5%)
East Sterea Hellas 1/7 0/4 1/11 (9,1%)
Peloponnese & Crete 0/1 0/1 0/2 (0,0%)
TOTAL 6/19 (31,6%) 6/22 (27,3%) 12/42 (28,6%)

In the previous published study of 1969, antibodies were found in 20.8% of the
tested serum samples. It is necessary to point out that that study refers to swine blood
serum samples which were tested before the «industrialization» of pig farming and
before the onset of vaccination programs (the vaccinations for PRV in Greece started
in the mid 80’s), whereas in the present study, the majority of the farms (75%) is
practicing a vaccination scheme against PRV. The latter finding indicates that
vaccination alone is not sufficient to eradicate the disease, unless it is accompanied
by other measures such as the removal of the animals, which are found positive to the
presence of antibodies against PRV. The proportion of positive PRV farms for the
level of each predictor is indicated in table 3. The analysis of the data presented in
table 3 showed that 83.3% of the PRV infected farms were purchasing breeding
animals (gilts or/and boars) from sources outside the farm and 100% of them did not
apply quarantine for the newly purchased breeding animals!

4
Table 3. Characteristics of the sampled breeding farms in Greece (42 farms) and descriptive
statistics for predictors tested for association with PRV positive status

Number Characteristic
Farms
Predictors Category (%) of positive within positive
(number)
sites (%) herds (%)

Size (No sows) <300 22 52,4 6 (27,3) 50,0
≥300 20 47,6 6 (30,0)
Density
<20 35 83,3 7 (20) 53,8
(farms/1000 km2)
≥20 7 16,7 5 (71,4)
Distance (km) <6 28 66,7 11 (39,3) 91,7
≥6 14 33,3 1 (7,1)
Gilt purchase No 13 31,0 2 (15,4)
Yes 29 69,0 10 (34,5) 83,3
Quarantine No 32 78,0 12 (37,5) 100,0
Yes 9 22,0 0 (0,0)
Biosecurity
No 19 45,2 10 (52,6) 83,3
measures
Yes 23 54,8 2 (8,7)
AIAO No 18 43,9 8 (44,4) 66,7
Yes 23 56,1 4 (17,4)
PRV-vaccination No 7 16,7 3 (42,9)
Yes 35 83,3 9 (25,7) 75,0
Economic
No 14 33,3 3 (21,4)
problems
Yes 28 66,7 9 (32,1) 69,2
Mortality No 32 76,2 6 (18,8)
Yes 10 23,8 6 (60,0) 50,0
Nervous signs No 36 85,7 9 (25.0)
Yes 6 14,3 3 (50,0) 25,0
Respiratory signs No 16 38,1 2 (12,5)
Yes 26 61,9 10 (38,5) 83,3
Gastroenteric
No 27 64,3 7 (25,9)
signs
Yes 15 35,7 5 (33,3) 41,7
Reproductive
No 20 47,6 5 (25,0)
signs
Yes 22 52,4 7 (31,8) 58,3
Neonatal stage
No 39 92,9 9 (23,1)
problems
Yes 3 7,1 2 (66,7) 16,7
Nursery stage
No 13 31,0 2 (15,4)
problems
Yes 29 69,0 10 (34,5) 83,3
Grower stage
No 16 38,1 1 (6,3)
problems
Yes 26 61,9 11 (42,3) 91,7
Finisher stage
No 29 69,0 5 (17,2)
problems
Yes 13 31,0 7 (53,8) 58,3

Moreover, according to the results of the multivariable logistic regression analysis,
it appears that factors such as “pig herd area density” and “hygienic/ biosecurity
measures” play a key role in the probability of a farm to become PRV positive. More

5
specifically, farms which were located in low-density areas and were applying
hygienic/biosecurity measures, had a predicted probability of being positive for PRV
of 1.97%. However, their probability was increased to 26.7% when farms were
located in low-density areas but were not applying hygienic/biosecurity measures or
to 30.8% when farms were located in high-density areas and were applying
hygienic/biosecurity measures. Finally, the probability of being positive for PRV was
increased to 88.9% when farms were located in high-density areas and were not
applying hygienic/biosecurity measures. It is obvious that the higher the density of
pig farms of an area, the more likely is that a farm will become positive for PRV.
Factors such as movement of flying insects, as well as of other wild and domestic
animals should play a role in the spread of PRV in high-density areas.
MCA had shown (table 4) that the presence of wild-type PRV is mostly related
with problems in growing and finishing pigs, as well as is also related to respiratory
and reproductive disease problems and increased mortality. Lack of appropriate
biosecurity measures and economical problems may be substantial factors for such
presence.

Table 4. Multiple correspondence analysis (MCA)

CTR F1
37 -283 D32 Problems in growing pigs 6
37 -418 A11 Presence of PRV 7
34 -273 C42 Presence of respiratory problems 8
34 -386 D42 Problems in finishing pigs 9
29 -408 C22 Increased mortality 13
28 -283 B81 Abscence of biosecurity measures 14
23 -247 C62 Presence of reproductive problems 17
22 -196 C12 Presence of general health problems 19

22 -209 B52 Presence of economical problems 20
21 -225 A41 Presence of A. pleuropneumoniae 21

In conclusion, this study provides new information regarding the presence of PRV
in Greek pig farms. The use of such information may assist in designing and
implementing measures to control and eradicate the disease from Greece.

References
1. Hahn, E.C., Fadl-Alla, B. and Lichtensteiger, C.A. (2010) Variation of
Aujeszky’s disease viruses in wild swine in USA Vet. Microbiol. 143, 45-51.
2. MacDiarmid, S.C. (2000) Aujeszky’s disease eradication in New Zealand. Aust.
Vet. J. 78, 470-471.

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3. Marinou, K.A., Papatsiros, V.G., Gkotsopoulos, E.K., Odatzoglou, P.K. and
Athanasiou, L.V. (2015) Exposure of extensively farmed wild boars (Sus scrofa
scrofa) to selected pig pathogens in Greece. Vet. Q. 35(2), 97-101.
4. Μeng, X.J., Lindsay, D.S. and Sriranganathan, N. (2009) Wild boars as sources
for infectious diseases in livestock and humans. Philos. Trans. R. Soc. Lond. B.
Biol. Sci. 364, 2697–2707.
5. Mettenleiter, T.C., Ehlers, B., Muller, T., Yoon K-J. and Teifke, J.P. (2012)
Herpesviruses. In Diseases of swine 10th edition, 421-446.
6. Papatsas, J., Paschaleri-Papadopoulou, E., Koubati-Artopiou, M., Kritas, S.K.
and Kyriakis, S.C. (1995) Aujeszky's Disease in pigs: Update review and
proposed measures for its control and eradication in Greece. Journal of Hellenic
Veterinary Medical Society 46,19-29.
7. Touloudi, A., Valiakos, G., Athanasiou, L.V., Birtsas, P., Giannakopoulos, A.,
Papaspyropoulos, K., Kalaitzis, C., Sokos, C., Tsokana, C.N., Spyrou, V.,
Petrovska, L. and Billinis, C. (2015) A serosurvey for selected pathogens in
Greek European wild boar. Vet. Rec. Open 2:e000077. Doi:10.1136/vetreco-
2014-000077.