<!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Archiving and Interchange DTD v1.0 20120330//EN" "JATS-archivearticle1.dtd">
<article xmlns:xlink="http://www.w3.org/1999/xlink">
  <front>
    <journal-meta />
    <article-meta>
      <title-group>
        <article-title>Epidemiological Investigation of Pseudorabies in Greece</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <string-name>Konstantinos Papageorgiou</string-name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Evanthia Petridou</string-name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Georgios Filioussis</string-name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Alexandros Theodoridis</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Ioannis Grivas</string-name>
          <xref ref-type="aff" rid="aff3">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Odysseas Moshidis</string-name>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Spyridon K. Kritas</string-name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <aff id="aff0">
          <label>0</label>
          <institution>Animal Production Economics</institution>
        </aff>
        <aff id="aff1">
          <label>1</label>
          <institution>Department of Business Administration, University of Macedonia</institution>
          ,
          <addr-line>Thessaloniki</addr-line>
          ,
          <country country="GR">Greece</country>
        </aff>
        <aff id="aff2">
          <label>2</label>
          <institution>Department of Microbiology and Infectious Diseases</institution>
        </aff>
        <aff id="aff3">
          <label>3</label>
          <institution>Laboratory of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, School of Health Sciences, Aristotle University of Thessaloniki</institution>
        </aff>
      </contrib-group>
      <abstract>
        <p>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.</p>
      </abstract>
      <kwd-group>
        <kwd>pseudorabies</kwd>
        <kwd>epidemiology</kwd>
        <kwd>pigs</kwd>
        <kwd>eradication</kwd>
      </kwd-group>
    </article-meta>
  </front>
  <body>
    <sec id="sec-1">
      <title>1 Introduction</title>
      <p>
        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)
        <xref ref-type="bibr" rid="ref6">(Mettenleiter et al., 2012)</xref>
        .
      </p>
      <p>
        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
        <xref ref-type="bibr" rid="ref1">(Hahn et
al., 2010)</xref>
        . PRV has also been eradicated from Canada, New Zealand and USA
        <xref ref-type="bibr" rid="ref2">(MacDiarmid, 2000)</xref>
        . 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
source of infection for domestic pigs. In countries that are free of PRV, vaccination is
prohibited.
      </p>
      <p>
        Greece belongs to the countries where the disease is still enzootic. According to an
old serological study in 1969
        <xref ref-type="bibr" rid="ref7">(Papatsas et al., 1995)</xref>
        , 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
        <xref ref-type="bibr" rid="ref4 ref8">(Touloudi et al., 2015; Marinou et al.,
2015)</xref>
        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
      </p>
    </sec>
    <sec id="sec-2">
      <title>Materials and Methods</title>
      <p>The study was carried out in Greece from October 2010 to October 2011.
Fortytwo (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.</p>
      <p>Number of farms sampled /Number of
farms in territory</p>
      <p>Herd-size category
Small
4/16
4/18
4/81
2/66
7/21
1/25</p>
      <p>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:
•
•
•
•
•
•
•
•
•
•
•
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 (&lt;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 (&lt;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).</p>
      <p>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
antiPRV-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.</p>
      <p>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.</p>
    </sec>
    <sec id="sec-3">
      <title>3 Results – Discussion</title>
      <p>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).</p>
      <p>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!
47,6
92,9
7,1
31,0
69,0
38,1
61,9
69,0
31,0</p>
      <p>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
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.</p>
      <p>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.</p>
      <p>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.</p>
    </sec>
  </body>
  <back>
    <ref-list>
      <ref id="ref1">
        <mixed-citation>
          <string-name>
            <surname>Hahn</surname>
            ,
            <given-names>E.C.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Fadl-Alla</surname>
            ,
            <given-names>B.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Lichtensteiger</surname>
            ,
            <given-names>C.A.</given-names>
          </string-name>
          (
          <year>2010</year>
          )
          <article-title>Variation of Aujeszky's disease viruses in wild swine in USA Vet</article-title>
          . Microbiol.
          <volume>143</volume>
          ,
          <fpage>45</fpage>
          -
          <lpage>51</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref2">
        <mixed-citation>
          <string-name>
            <surname>MacDiarmid</surname>
            ,
            <given-names>S.C.</given-names>
          </string-name>
          (
          <year>2000</year>
          )
          <article-title>Aujeszky's disease eradication in New Zealand</article-title>
          . Aust.
        </mixed-citation>
      </ref>
      <ref id="ref3">
        <mixed-citation>
          <string-name>
            <surname>Vet</surname>
          </string-name>
          . J.
          <volume>78</volume>
          ,
          <fpage>470</fpage>
          -
          <lpage>471</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref4">
        <mixed-citation>
          3.
          <string-name>
            <surname>Marinou</surname>
            ,
            <given-names>K.A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Papatsiros</surname>
            ,
            <given-names>V.G.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Gkotsopoulos</surname>
            ,
            <given-names>E.K.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Odatzoglou</surname>
            ,
            <given-names>P.K.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Athanasiou</surname>
            ,
            <given-names>L.V.</given-names>
          </string-name>
          (
          <year>2015</year>
          )
          <article-title>Exposure of extensively farmed wild boars (Sus scrofa scrofa) to selected pig pathogens in Greece</article-title>
          . Vet. Q.
          <volume>35</volume>
          (
          <issue>2</issue>
          ),
          <fpage>97</fpage>
          -
          <lpage>101</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref5">
        <mixed-citation>
          4.
          <string-name>
            <surname>Μeng</surname>
            ,
            <given-names>X.J.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Lindsay</surname>
            ,
            <given-names>D.S.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Sriranganathan</surname>
            ,
            <given-names>N.</given-names>
          </string-name>
          (
          <year>2009</year>
          )
          <article-title>Wild boars as sources for infectious diseases in livestock and humans</article-title>
          .
          <source>Philos. Trans. R. Soc. Lond. B. Biol. Sci</source>
          .
          <volume>364</volume>
          ,
          <fpage>2697</fpage>
          -
          <lpage>2707</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref6">
        <mixed-citation>
          5.
          <string-name>
            <surname>Mettenleiter</surname>
            ,
            <given-names>T.C.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Ehlers</surname>
            ,
            <given-names>B.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Muller</surname>
            ,
            <given-names>T.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Yoon</surname>
            <given-names>K-J.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Teifke</surname>
            ,
            <given-names>J.P.</given-names>
          </string-name>
          (
          <year>2012</year>
          )
          <article-title>Herpesviruses</article-title>
          .
          <source>In Diseases of swine 10th edition</source>
          ,
          <fpage>421</fpage>
          -
          <lpage>446</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref7">
        <mixed-citation>
          6.
          <string-name>
            <surname>Papatsas</surname>
            ,
            <given-names>J.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Paschaleri-Papadopoulou</surname>
            ,
            <given-names>E.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Koubati-Artopiou</surname>
            ,
            <given-names>M.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Kritas</surname>
            ,
            <given-names>S.K.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Kyriakis</surname>
            ,
            <given-names>S.C.</given-names>
          </string-name>
          (
          <year>1995</year>
          )
          <article-title>Aujeszky's Disease in pigs: Update review and proposed measures for its control and eradication in Greece</article-title>
          .
          <source>Journal of Hellenic Veterinary Medical Society</source>
          <volume>46</volume>
          ,
          <fpage>19</fpage>
          -
          <lpage>29</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref8">
        <mixed-citation>
          7.
          <string-name>
            <surname>Touloudi</surname>
            ,
            <given-names>A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Valiakos</surname>
            ,
            <given-names>G.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Athanasiou</surname>
            ,
            <given-names>L.V.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Birtsas</surname>
            ,
            <given-names>P.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Giannakopoulos</surname>
            ,
            <given-names>A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Papaspyropoulos</surname>
            ,
            <given-names>K.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Kalaitzis</surname>
            ,
            <given-names>C.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Sokos</surname>
            ,
            <given-names>C.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Tsokana</surname>
            ,
            <given-names>C.N.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Spyrou</surname>
            ,
            <given-names>V.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Petrovska</surname>
            ,
            <given-names>L.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Billinis</surname>
            ,
            <given-names>C.</given-names>
          </string-name>
          (
          <year>2015</year>
          )
          <article-title>A serosurvey for selected pathogens in Greek European wild boar</article-title>
          .
          <source>Vet. Rec. Open</source>
          <volume>2</volume>
          :
          <fpage>e000077</fpage>
          . Doi:
          <volume>10</volume>
          .1136/vetreco2014-
          <fpage>000077</fpage>
          .
        </mixed-citation>
      </ref>
    </ref-list>
  </back>
</article>