Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae (A.pleuropneumoniae) is one of the most important respiratory diseases of pigs and causes worldwide severe losses in pig farming. For A.pleuropneumoniae control and monitoring, the detection of ApxIV antibodies in the serum is the most frequently used serological method. The aim of this study was to investigate presence of antibodies against A. pleuropneumoniae in blood sera of gilts and sows using the ELISA test. Samples were taken from gilts and sows originating from four commercial swine farms in Serbia. For detection of ApxIV antibodies, commercial ELISA kit was used. A total of 453 blood sera samples of gilts (207) and sows (246) were examined. Antibodies against A. pleuropneumoniae were detected in 57 (12.58%) sera. Antibodies were present in 22 (10.62 %) sera of gilts and in 35(14.22%) sera of sows. Percentage of positive sera differed among the farms, ranging in gilts from 3.33-17.77 % and in sows from 8.95-22.64%. Serological methods is one of the most important procedures in the diagnosis of porcine pleuropneumonia particularly suitable for the control of animal health status in a large breeding

Radosavljević V.

Vojinović D.

Žutić Jadranka

Biotechnology in Animal Husbandry

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Biotechnology in Animal Husbandry 30 (2), p 295-302,  2014                                   ISSN 1450-9156 
Publisher: Institute for Animal Husbandry, Belgrade-Zemun                               UDC 636.09'636.4
DOI: 10.2298/BAH1402295Z 
 
   
 
SEROPREVALENCE OF ACTINOBACILLUS 
PLEUROPNEUMONIAE IN SWINE ORIGINATED FROM 
COMMERCIAL FARMS IN SERBIA  
 
J. Žutić, D. Vojinović, V. Radosavljević 
 
1Institute of Veterinary Medicine of Serbia, Vojvode Toze 14, 11000 Belgrade, Serbia 
Corresponding author: jadranka.zutic@gmail.com 
Original scientific paper 
 
Abstract: Porcine pleuropneumonia caused by Actinobacillus 
pleuropneumoniae (A.pleuropneumoniae)  is one of the most important respiratory 
disease of pigs and causes worldwide severe losses in pig farming. For 
A.pleuropneumoniae control and monitoring, the detection of ApxIV antibodies in 
the serum is the most frequently used serological method. The aim of this study 
was to investigate presence of antibodies against A. pleuropneumoniae in blood 
sera of gilts and sows using the ELISA test. Samples were taken from gilts and 
sows originating from four commercial swine farms in Serbia. For detection of 
ApxIV antibodies, commercial ELISA kit was used. A total of 453 blood sera 
samples of gilts (207) and sows (246) were examined. Antibodies against A. 
pleuropneumoniae were detected in 57 (12.58%) sera. Antibodies were present in 
22 (10.62 %) sera of gilts and in 35(14.22%) sera of sows. Percentage of positive 
sera differed among the farms, ranging in gilts from 3.33-17.77 % and in sows 
from 8.95-22.64%. Serological methods is one of the most important procedures in 
the diagnosis of porcine pleuropneumonia particularly suitable for the control of 
animal health status in a large breeding.  
 
Key words: Actinobacillus pleuropneumoniae, gilt, sow, antibodies, 
ELISA. 
 
Introduction   
 
        The most significant problems in contemporary pig production are in 
connection  with diseases of the respiratory system (Baker,2005; Hansen et al., 
2010). It is a characteristic of the current manner of production to set up 
agglomerations with concentrations of large numbers of animals within a small 
space. As a multifactorial disease, environmental conditions, population size, 
management strategies and pig-specific factors such as age and genetics also play 
significant roles in the outcome of respiratory disease (Opriessing et al., 2011).  
J. Žutić et al. 
 
 
 
296
Such conditions are especially favourable for respiratory pathogens and continuous 
presence of a high degree of virulence in vivo. As a result, there are increasingly 
frequent outbreaks of respiratory infections which are more difficult to control, 
with the maintenance conditions in large agglomerations exerting an extremely 
unfavorable effect (Žutić et al., 2009). The prevalence of respiratory disease is 
affected by the following: the presence of respiratory pathogenic organisms, the 
virulence of the pathogens present, the level of the pathogens in the house 
environment, the immunity of the pig and the time of exposure to the organisms, 
the presence of secondary opportunistic bacteria, the interactions between 
management, environment, the diseases and the pig (The Pig Site Pig 
Health,2013).                                                           
        Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae 
(A.pleuropneumoniae) is one of the most important respiratory disease of pigs and 
is widely spread among pig-keeping countries (Vidić et al., 2004; Bochev, 2007; 
Vaduva et al., 2011). A. pleuropneumoniae is a small, gram-negative, encapsulated 
rod belonging to the family Pasteurellaceae. 
         There are two biotypes of A.pleuropneumoniae, differentiated on the basis of 
their requirement for factor V, ( NAD) in biotype I (NAD-dependent) and biotype 
II (NAD-independent). There are 13 serotypes of biotype I and 2 serotypes of 
biotype II , based on surface polysaccharide antigens (Maldonado et al., 2009; 
Bessone, 2010; Gotschalk, 2012). The four major RTX toxins ApxI, ApxII, ApxIII 
and ApxIV are secreted by the different serotypes in various combinations (Shin et 
al.,2010). All serotypes can secrete ApxIV which can be produced only during 
infection, but not in vitro conditions. Presence of the Apx toxins is strongly 
correlated with virulence. 
            Growing pigs are most likely to be affected when they are 12-16 weeks old, 
but the disease can occur in all ages of swine (Gotschalk and Taylor, 2006). This 
disease is causing significant loss to farmers particularly when finisher pigs are 
involved. Pigs, asymptomatically infected with A.pleuropneumoniae in their upper 
respiratory tract, can transmit the infection.  The main route of spread is by direct 
contact from pig to pig or by aerosol within short distances, although some authors 
reported that airborne transmission between closely located pig units is possible but 
rare (Woeste and Grosse, 2007). The clinical course of the disease can vary widely, 
ranging from the acute forms with severe clinical signs and a high mortality to the 
more chronic forms with few or even without any clinical symptoms (Shi et al., 
2012). In the absence of treatment, the disease can progress very rapidly and death 
can occur within a few hours. Main clinical signs of the acute disease are anorexia, 
depression, fever, dyspnea and/or polypnea. Chronic infections are characterized 
by coughing and pleuritis in lungs (Gotschalk et al., 2010).  The recent 
epidemiological studies indicate a very high rate of exposure reaching up to 100% 
of seropositivity of investigated farms (Krejci and Newberry, 2011). Confirmation 
by laboratory testing is essential for diagnosis, especially in monitoring schemes. 
Seroprevalence of actinobacillus... 
 
 
 
297 
The health monitoring of herds is extremely important, firstly because of the need 
for the adequate strategy to be chosen for controlling the porcine pleuropneumonia 
and, at the same time, in order to prevent economic losses that this disease may 
cause. Identification of seropositive animals is an important measure to control and 
eliminate the disease from the swine farms. Also, serological tests showed 
seroconversion of each animal according to certain technological categories within 
the herd. Using ELISA, it is possible to detect ApxIV antibodies against all 
serotypes of the A. pleuropneumoniae, without cross-reaction with other bacterial 
species (Dreyfus et al., 2004; Nussbaumer et al., 2008; Eamens et al., 2012). The 
aim of the study was to investigate the presence of ApxIV antibodies against A. 
pleuropneumoniae in gilts and sows in four pig farms in Serbia.  
 
Material and methods 
 
        For A.pleuropneumoniae monitoring purposes, the detection of ApxIV 
antibodies in the serum is currently the most frequently used serological method. 
ELISA test can be used to evaluate the A. pleuropneumoniae status of commercial 
herds  especially for the diagnosis of latently infected, without clinical signs. For 
the investigations, samples were taken of the blood of 207 gilts and 246 sows 
originating from 4 commercial swine farms in Serbia. The capacity of each farm is 
about 700-1.000 sows. Investigations were carried out using the method of ELISA 
with the following diagnostic kits: Chekit APP-Apx IV: A. pleuropneumoniae 
antibody test Kit 
 (IDEXX APP-ApxIV Ab Test).         
 
Results and Discussions 
 
A total of 453 blood sera samples of gilts (207) and sows (246) were 
examined. Results of determination of antibodies against of A. pleuropneumoniae 
are given in Table 1. 
The results of the investigations have shown that infection with 
A.pleuropneumoniae is present on all four examined farms. Antibodies against A. 
pleuropneumoniae were detected in 57 (12.58%) sera. Antibodies were present in 
22 (10.62%) sera of gilts and in 35 (14.22%) sera of sows. Percentage of positive 
sera differed among the farms, ranging in gilts from 3.33-17.77% and in sows 8.95-
22.64%. The results of this study show a significant decrease of seropositive 
animals. In previous study, antibodies against A. pleuropneumoniae were detected 
in 69.86% of sows and 76.72% of gilts (Žutić J, et al.,2008).   
 
 
 
J. Žutić et al. 
 
 
 
298
Table 1. Results on presence of antibodies against Actinobacillus pleuropneumoniae in gilts and 
sows blood sera 
 
      Farms Category No. investigated     No. positive    % positive 
gilts 49 7 14.28  
1 sows 55 9 16.36 
gilts 53 5 9.43  
2 sows 61 8 13.11 
gilts 60 2 3.33  
3 sows 67 6 8.95 
gilts 45 8 17.77  
4 sows 63 12 22.64 
Total gilts  207 22 10.62 
Total sows   246 35 14.22 
Total animals  453 57 12.58 
  
It is probably result of intensive serological monitoring of gilts prior to 
fertilization and improving the living conditions of animals. A higher percentage of 
positive animals on the farm 4 are the result of old buildings and poor conditions. 
Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae is 
still a problem today in herds of many countries where the swine industry is 
important (Gotschalk et al.,2010).The recent epidemiological studies indicate a 
very high rate of exposure, reaching up to 100% of seropositivity of investigated 
farms. Of those,  90% in Northwest Germany, 96% in Belgium, 89% in Spain and 
100% in Italy and Belarus (Krejci and Newberry,2011). In Belgium, 
seroprevalence for serovar 2 was 58% (range 0 to 100 %), 53% for serovar 3 (range 
10 to 95%), and 35% for serovar 9 (range 5 to 100%) (Maes et al., 2002). In the 
first report of the presence of A. pleuropneumoniae in Turkey, 258 out of 384 
blood samples (67 %) were positive (Metiner,2007). Shi (Shi et all, 2012) reports 
of  55.72% seropositive of the Tibetan pigs. The prevalence ranged from 42.68-
71.11%. This agrees with Assavacheep (Assavacheep et al., 2003), who had 
reported that  60% pigs were seropositive to at least one serotype and 45% of the 
pigs were seropositive to more than one serotype.   
        The successful control of porcine pleuropneumonia depends on the efficiency 
of preventing intra- and inter-farm transmission of the infection. One of the ways to 
successfully control and eliminate the porcine pleuropneumonia is a timely and fast 
diagnostic procedure with the implementation of immunodiagnostic tests (Žutić et 
al., 2008). 
      Serological diagnostics of infections caused by A.pleuropneumoniae is 
essential measure for the identification of latently infected herds and the 
determination of multiplicity of serotypes within the herd. The serological control 
Seroprevalence of actinobacillus... 
 
 
 
299 
of gilts is of particular importance to detect infected animal prior to fertilization, 
because the mothers can transmit the infection to their offspring at birth. 
 
Conclusions 
 
It is very important to identify the presence of individual agents in the 
etiopathogenesis of respiratory disease of pigs in a large agglomerations. Based on 
our results, ELISA is a reliable method for serological diagnosis of porcine 
pleuropneumonia. This method is specific and may be used for the routine 
surveillance of health status in pig herds.  
 
Acknowledgements 
This investigation was supported by a grant from the Ministry of Education 
and Science of the Republic of Serbia, Project No. 31079. 
 
Seroprevalencija Actinobacillus pleuropneumoniae  kod  
svinja  poreklom  sa komercijalnih  farmi u Srbiji 
 
J. Žutić, D. Vojinović, V.r Radosavljević 
 
Rezime 
 
Pleuropneumonija svinja jedna je od najznačajnijih respiratornih bolesti 
svinja koja širom sveta dovodi do gubitaka, posebno u intenzivnoj svinjarskoj 
proizvodnji. Uzročnik bolesti je A.pleuropneumoniae.Prema korištenju β-
nikotinamid adenin dinukleotida (β-NAD) za rast, uzročnik je podeljen na biovar 1 
(β-NAD ovisan) i biovar 2 (β-NAD neovisan). Svaki biovar je dalje podeljen na 
serotipove. Do sada je otkriveno 15 različitih serotipova, među kojima su 
ustanovljene znatne razlike u virulenciji ali isto tako i varijacije u virulenciji 
između sojeva istog serotipa.Glavne faktore virulencije predstavljaju toksini.Sojevi 
A.pleuropneumoniae stvaraju četiri tipa egzotoksina označenih ApxI, ApxII, 
ApxIII i ApxIV. Za razliku od ostalih, ApxIV toksin proizvode svi serotipovi i 
visoko je specifičan za A.pleuropneumoniae.Bolest se klinički manifestuje kašljem, 
teškim disanjem i visokom temperaturom, a patoanatomski hemoragično-
nekrotičnim promenama na plućima.U brojnim je istraživanjima dokazano da ovaj 
patogen često učestvuje u interakcijama bilo sa  bakterijskim ili virusnim 
patogenima kao što su Mycoplasma hyopneumoniae, PRRSV i PCV2.Glavni put 
širenja je direktni kontakt među svinjama ili aerosolom na kratkoj distanci kao i 
mogućnost prenosa uzročnika sa majki na prasad.U cilju praćenja pojave i kontrole 
pleuropneumonije, najčešće se koriste serološke metode koje otkrivaju prisustvo 
J. Žutić et al. 
 
 
 
300
antitela za ApxIV toksin A.pleuropneumoniae.Cilj je ovog rada bio da se ispitaju 
krvni serumi nazimica i krmača poreklom sa 4 svinjarske farme u Srbiji,na 
prisustvo specifičnih antitela za A.pleuropneumoniae. U ispitivanju je, za 
otkrivanje antitela korišten komercijalni ELISA kit (IDEXX APP-ApxIV Ab Test). 
Ukupno je serološki pregledano 453 uzorka krvnih seruma, i to 207 uzoraka od 
nazimica i 246 uzoraka od krmača. Antitela za A.pleuropneumoniae ustanovljena 
su u 57 (12,58%) od ukupno 453 pregledanih seruma životinja. Kod nazimica, 
antitela su ustanovljena u 22 (10,62%) a kod krmača u 35(14,22%) 
seruma.Procenat pozitivnih seruma životinja bio je različit među farmama i kretao 
se kod nazimica od 3,33-17,77 % a kod krmača 8,95-22,64%. U dijagnostici 
pleuropneumonije svinja, serološke metode predstavljaju jedan od najznačajnijih 
postupaka, posebno pogodnih za kontrolu zdravstvenog stanja životinja u velikim 
aglomeracijama, pri čemu je moguće otkriti i latentno inficirana grla. 
 
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Received 20 March 2014; accepted for publication 4 June 2014 
 
 
 


Seroprevalence of Actinobacillus pleuropneumoniae in swine originated from commercial farms in Serbia

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Seroprevalence of Actinobacillus pleuropneumoniae in swine originated from commercial farms in Serbia

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